Method For Treating Dissolving Pulp

Abstract

The present invention relates to treatment of dissolving pulp with a lytic polysaccharide monooxygenase. The lytic polysaccharide monooxygenase treatment results in reduced viscosity and/or improved viscosity control in the dissolving pulp production process and/or 5 increased reactivity of the dissolving pulp.

Description

REFERENCE TO SEQUENCE LISTING

[0001] This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference.

FIELD OF THE INVENTION

[0002] The present invention relates to treatment of dissolving pulp with one or more enzymes. The enzymatic treatment results in reduced viscosity and/or improved viscosity control in the dissolving pulp production process and/or increased reactivity of the final dissolving pulp.

BACKGROUND OF THE INVENTION

[0003] Commercial dissolving pulp or dissolving-grade pulp is a chemical bleached pulp with a high cellulose content enough to be suitable for the production of regenerated cellulose and cellulose derivatives. Commercial dissolving pulp has special properties, such as a high level of brightness and uniform molecular-weight distribution. Commercial dissolving pulp is manufactured for uses that require a high chemical cellulose purity, and particularly low hemicellulose content, since the chemically similar hemicellulose can interfere with subsequent processes. Dissolving pulp is so named because it is not made into paper, but dissolved either in a solvent or by derivatization into a homogeneous solution, which makes it completely chemically accessible and removes any remaining fibrous structure. Once dissolved, it can be spun into textile fibers such as viscose or Lyocell, or chemically reacted to produce derivatized celluloses, such as cellulose triacetate, a plastic-like material formed into fibers or films, or cellulose ethers such as methyl cellulose, used as a thickener.

[0004] Conventional viscose manufacturing which uses dissolving pulps as raw material requires improvement with respect to its environmental impact as well as its production costs. There is a need in the art to provide a method for treating dissolving pulp with reduced costs and less environmental impact.

[0005] The present invention provides a lytic polysaccharide monooxygenase-based solution that reduces the viscosity and/or improves the viscosity control in the production of dissolving pulp, e.g., kraft and sulfite dissolving pulp. The enzyme solution described in this invention allows a more selective depolymerization of cellulose and thus a better control of pulp viscosity as compared to the conventional methods in use that are unselective with many side reactions, such as oxygen, hydrogen peroxide, ozone, sodium hypochlorite and acid hydrolysis. Furthermore, the reactivity of the dissolving pulp in the present invention is improved, thereby reducing the amount of chemicals used in the viscose production process and/or improving the processability in terms of viscose dope filterability in the viscose making process. Savings in the amount of chemicals utilized in the production of regenerated cellulose such as carbon disulfide (CS.sub.2) in the viscose making process will reduce costs and environmental impact. Similarly, the reactivity increase of the dissolving pulp is expected to benefit the production process of cellulose derivatives, such as subsequent esterification and etherification processes.

SUMMARY OF THE INVENTION

[0006] The invention provides a method for treating dissolving pulp, comprising a step of subjecting the dissolving pulp to a lytic polysaccharide monooxygenase.

[0007] The method of the present invention generates dissolving pulp with reduced viscosity and/or improved viscosity control in the dissolving pulp production process, and/or increased reactivity for viscose making, and/or increased content of oxidized groups, compared to dissolving pulp obtained by the same process where the lytic polysaccharide monooxygenase (LPMO) treatment is omitted. The said dissolving pulp is kraft dissolving pulp and/or sulfite dissolving pulp.

[0008] The present invention further provides a dissolving pulp made by the method of the present invention.

[0009] The present invention further provides a textile fiber or a derivatized cellulose made of the dissolving pulp of the present invention.

[0010] The present invention further provides use of a lytic polysaccharide monooxygenase for treatment of dissolving pulp.

[0011] Definitions

[0012] Dissolving pulp: Dissolving pulp is a high-grade cellulose pulp, with low contents of hemicellulose, lignin and resin. This pulp has special properties, such as high level of brightness and uniform molecular weight distribution. It is used to make products that include rayon and acetate textile fibers, cellophane, photographic film and various chemical additives. To a large extent, use of dissolving wood pulp depends on its purity (cellulose content), which depends mainly on the production process. To obtain products of high quality, these so-called "special" pulps must fulfill certain requirements, such as high cellulose content, low hemicellulose content, a uniform molecular weight distribution, and high cellulose reactivity. Most of the commercial dissolving pulps accomplish these demands to a certain extent. Nevertheless, achieving high cellulose accessibility as well as solvent and reagent reactivity is not an easy task due to the compact and complex structure presented by the cellulose. About 77% of all dissolving pulp is used in the manufacture of cellulosic fibers (rayon and acetate).

[0013] Two basic processes are used to produce dissolving pulp: (a) the sulfite process; and b) the sulfate process (kraft).

[0014] To manufacture dissolving-grade pulps, removing hemicelluloses from the wood fiber is crucial, because hemicelluloses can affect the filterablility of viscose, the xanthation of cellulose and the strength of the end product during the production of viscose. Hemiceluloses are removed during the cooking of wood and the subsequent bleaching. In sulfite pulping, the acidic conditions used are responsible for removing most of the hemicellulose while in sulfate/kraft process usually a prehydrolysis step is required to remove hemicelluloses. Another method to remove hemicelluloses is by treatment of pulps with enzymes that react only with the hemicellulose portion of the pulp.

[0015] Kraft dissolving pulp: "Kraft dissolving pulp" is synonymous with "sulphate dissolving pulp". A preferred example is a prehydrolysis kraft dissolving pulp. Kraft dissolving pulp is produced by digesting wood chips at temperatures above about 120.degree. C. with a solution of sodium hydroxide and sodium sulfide. Some kraft pulping is also done in which the sodium sulfide is augmented by oxygen or anthraquinone. As compared with soda pulping, kraft pulping is particularly useful for pulping of softwoods, which contain a higher percentage of lignin than hardwoods. The term "kraft dissolving pulp" is synonymous with "kraft dissolving cellulose" and "kraft dissolving-grade pulp" and refers to pulp that has a high cellulose content. The cellulose content of the kraft dissolving pulp is preferably at least 90% (weight/weight) such as at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% (w/w). Kraft dissolving pulp is manufactured for uses that require a high chemical purity, and particularly low hemicellulose content. The hemicellulose content of the dissolving pulp is preferably less than 10% (weight/weight) such as less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1% (w/w). Kraft dissolving pulp can e.g. be used for generation of regenerated cellulose or for generation of cellulose derivatives. "Kraft dissolving-grade pulp" can also be defined as pulp that has been purified sufficiently for use in the production of viscose, rayon, cellulose ethers, or cellulose esters with organic or inorganic acids.

[0016] Sulfite dissolving pulp: The sulfite process produces wood pulp which is almost pure cellulose fibers by using various salts of sulfurous acid to extract the lignin from wood chips in large pressure vessels called digesters. The salts used in the pulping process are either sulfites (SO.sub.3.sup.2-), or bisulfites (HSO.sub.3.sup.-), depending on the pH. The counter ion can be sodium (Na.sup.+), calcium (Ca.sup.2+), potassium (K.sup.+), magnesium (Mg.sup.2+) or ammonium (NH.sup.4+).

[0017] Sulfite pulping is carried out between pH 1.5 and 5, depending on the counterion to sulfite (bisulfite) and the ratio of base to sulfurous acid. The pulp is in contact with the pulping chemicals for 4 to 14 hours and at temperatures ranging from 130 to 160.degree. C. (266 to 320.degree. F.), again depending on the chemicals used.

[0018] Most of the intermediates involved in delignification in sulfite pulping are resonance-stabilized carbocations formed either by protonation of carbon-carbon double bonds or acidic cleavage of ether bonds which connect many of the constituents of lignin. It is the latter reaction which is responsible for most lignin degradation in the sulfite process. The sulfite process is not expected to degrade lignin to the same extent that the kraft process does and the lignosulfonates from the sulfite process are useful byproducts.

[0019] The spent cooking liquor from sulfite pulping is usually called brown liquor, but the terms red liquor, thick liquor and sulfite liquor are also used (compared to black liquor in the kraft process). Pulp washers, using countercurrent flow, remove the spent cooking chemicals and degraded lignin and hemicellulose.

[0020] "Bleaching" is the removal of color from pulp, primarily the removal of traces of lignin which remains bound to the fiber after the primary pulping operation. Bleaching usually involves treatment with oxidizing agents such as chlorine (C-stage), chlorine dioxide (D-stage), oxygen (O-stage), hydrogen peroxide (P-stage), ozone (Z-stage) and peracetic acid (CH.sub.3CO.sub.3H; Paa-stage) or a reducing agent such as sodium dithionite (Y-stage). There are chlorine (Cl.sub.2; C-stage) free processes such as the elemental chlorine free (ECF) bleaching where chlorine dioxide (ClO.sub.2; D-stage) is mainly used and typically followed by an alkaline extraction stage. Totally chlorine free (TCF) bleaching is another process where mainly oxygen-based chemicals are used. The pulp bleaching process thus typically comprise a sequence of bleaching steps with washing in between them to remove the degradation products arising from the bleaching reactions.

[0021] Cold Caustic Extraction (CCE): A cold alkali extraction, also called Cold Caustic Extraction (CCE), is a method used to to remove short-chain noncellulosic carbohydrates (cellulose purification) that is based on physical effects such as swelling and solubilization. Usually, a CCE stage takes place at temperatures below 45.degree. C. and using very high NaOH dosage that, in the liquid phase, can reach values up to 100 g/L. Depending on the pulp consistency in use, this will determine the amount of NaOH per dry weight of pulp. Typical conditions for a CCE-stage can be 5-10% w/w NaOH in the liquid phase for at least 10 min.

[0022] Hot Caustic Extraction (HCE): the term "Hot Caustic Extraction" (HCE) is synonymous with "hot alkali extraction". HCE is a method to remove short chain hemicellulose and amorphous cellulose in pulps. A hot caustic extraction (HCE)-stage is a purification process that is based on chemical reactions, in particular alkaline peeling of hemicelluloses, which is carried out at higher temperatures and lower NaOH concentration compared to CCE.

[0023] ISO Brightness: ISO Brightness is defined in ISO 2470-1 (method for measuring ISO brightness of pulps, papers and boards), and it is the intrinsic radiance [reflectance] factor measured with a reflectometer having the characteristics described in ISO 2469.

[0024] Pulp viscosity: is measured by dissolving the pulp in a suitable cellulose solvent such as in cupri-ethylenediamine (CED) and measuring the solution viscosity. This measurement gives an indication of the average degree of polymerization of the cellulose. This property can be referred as intrinsic viscosity in mL/g and measured according to ISO 5351 or as TAPPI viscosity in cP and measured according to TAPPI T 230.

[0025] Unbleached or partially bleached or alkaline extracted kraft dissolving pulp: is produced by a kraft based cooking process such as pre-hydrolysis kraft (PHK) cooking but not fully bleached and purified until becoming a commercial kraft dissolving pulp and thus it is not a finished product. Typically it has an ISO brightness below 90% (such as below 85%, such as below 80%, such as below 75%, such as below 70%, such as below 65%, such as below 60%, such as below 55%, such as below 50%, such as below 45%, such as below 40%, such as below 35%, and such as below 30%).

[0026] Unbleached or partially bleached or alkaline extracted sulfite dissolving pulp: is produced by a sulfite based cooking process but not fully bleached and purified until becoming a commercial sulfite dissolving pulp and thus it is not a finished product. Typically it has an ISO brightness below 90% (such as below 85%, such as below 80%, such as below 75%, such as below 70%, such as below 65%, such as below 60%, such as below 55%, such as below 50%, such as below 45%, such as below 40%, such as below 35%, and such as below 30%).

[0027] Bleached kraft dissolving pulp and bleached sulfite dissolving pulp: is produced by a kraft dissolving pulp or a sulfite based cooking process but fully bleached and purified until becoming a commercial dissolving pulp. Typically it has an ISO brightness above 90% (such as above 91%, such as above 92%, such as above 93%, such as above 94%, such as above 95%, such as above 96%, such as above 97%, such as above 98%, such as above 99%, such as 100%).

[0028] Sequence identity: The relatedness between two amino acid sequences or between two nucleo-tide sequences is described by the parameter "sequence identity".

[0029] For purposes of the present invention, the sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EM-BOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably version 5.0.0 or later. The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) sub-stitution matrix. The output of Needle labeled "longest identity" (obtained using the--nobrief option) is used as the percent identity and is calculated as follows: (Identical Residues x 100)/(Length of Alignment Total Number of Gaps in Alignment)

DETAILED DESCRIPTION OF THE INVENTION

[0030] The invention relates to a method for treating dissolving pulp, comprising a step of subjecting the dissolving pulp to a lytic polysaccharide monooxygenase (LPMO).

[0031] In a preferred embodiment, the method of the present invention further comprises a step of subjecting the dissolving pulp to a cellulase.

[0032] In a preferred method of the present invention, the step of subjecting the dissolving pulp to a lytic polysaccharide monooxygenase and the step of subjecting the dissolving pulp to a cellulase are carried out simultaneously or sequentially in any order. In a further preferred embodiment of the present invention, a lytic polysaccharide monooxygenase is added to the dissolving pulp together with the cellulase. In a further preferred embodiment of the present invention, a lytic polysaccharide monooxygenase is added to the dissolving pulp before the addition of a cellulase. In a further preferred embodiment of the present invention, a lytic polysaccharide monooxygenase is added to the dissolving pulp after the addition of a cellulase.

[0033] In a preferred embodiment, the present method of the present invention comprises a step of bleaching the dissolving pulp. In a preferred method of the present invention, the step of subjecting the dissolving pulp to a lytic polysaccharide monooxygenase and the step of bleaching the dissolving pulp are carried out simultaneously or sequentially in any order.

[0034] In a preferred method of the present invention, the step of bleaching the dissolving pulp is performed using a chemical selected from the group consisting of ClO.sub.2, O.sub.2, O.sub.3, H.sub.2O.sub.2, CH.sub.3CO.sub.3H and NaOCl.

[0035] In a preferred embodiment, the method of the present invention further comprises the step of Alkaline Extraction. In the method of the present invention, Alkaline Extraction is an E, HCE or CCE stage. Special alkaline purification treatments such as HCE or CCE treatments can yield higher cellulose levels in sulfite and kraft processes. In the case of sulfite pulps, HCE is typically employed to further purify the pulp after the sulfite cooking.

[0036] In a preferred embodiment, the method of the present invention further comprises a step of subjecting the dissolving pulp to a lytic polysaccharide monooxygenase and an electron donor thereof, preferably ascorbic acid, gallic acid, pyrogallol or cysteine. The electron donor can exist in the dissolving pulp to be treated. In one embodiment, no or a little amount of electron donor is added to the dissolving pulp. In another embodiment, an effective amount of electron donor is added to the dissolving pulp.

[0037] In a preferred method of the present invention, the dissolving pulp is an unbleached, partially bleached, bleached or alkaline extracted dissolving pulp. In a preferred method of the present invention, the dissolving pulp is kraft pulp or sulfite pulp.

[0038] In one embodiment the lytic polysaccharide monooxygenase added in the method of the present invention has a sequence identity of at least 60% [such as at least 65%, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95%, such as at least 99%] to the mature polypeptide of SEQ ID NO: 1, the mature polypeptide of SEQ ID NO:2, the mature polypeptide of SEQ ID NO:3. In one embodiment the cellulase added in the method of the present invention has a sequence identity of at least 60% (such as at least 65%, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 90%, such as at least 95%, such as at least 99%) to SEQ ID NO: 4, the mature polypeptide of SEQ ID NO: 5, the mature polypeptide of SEQ ID NO: 6, or the mature polypeptide of SEQ ID NO: 7.

[0039] In a preferred embodiment, the lytic polysaccharide monooxygenase added in the method of the present invention comprises or consists of SEQ ID NO: 1 or the mature polypeptide thereof, or SEQ ID NO: 2 or the mature polypeptide thereof, or SEQ ID NO: 3 or the mature polypeptide thereof; or the cellulase comprises or consists of SEQ ID NO: 4, SEQ ID NO: 5 or the mature polypeptide thereof, SEQ ID NO: 6 or the mature polypeptide thereof, or SEQ ID NO: 7 or the mature polypeptide thereof.

[0040] In another preferred embodiment, the lytic polysaccharide monooxygenase added in the method of the present invention comprises amino acids 19 to 226 of SEQ ID NO: 1, or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the lytic polysaccharide monooxygenase added in the method of the present invention comprises amino acids amino acids 20 to 254 of SEQ ID NO: 2, or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the lytic polysaccharide monooxygenase added in the method of the present invention comprises amino acids 22 to 249 of SEQ ID NO: 3, or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the present invention comprises SEQ ID NO: 4 or the full length thereof, or a homologous sequence thereof, an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the present invention comprises amino acids 22-305 of SEQ ID NO: 5 or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the present invention comprises amino acids 22 to 293 of SEQ ID NO: 6 or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. In another preferred embodiment, the cellulase added in the method of the present invention comprises amino acids 19 to 409 of SEQ ID NO: 7 or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof.

[0041] The concentration of the lytic polysaccharide monooxygenase added in the method of the present invention is preferably from 0.05 mg/kg oven dry pulp to 100000 mg/kg oven dry pulp such as a concentration selected from the group consisting of from 0.05 mg/kg oven dry pulp to 250 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 1000 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 2000 mg/kg oven dry pulp, from 1.0 mg/kg oven dry pulp to 5000 mg/kg oven dry pulp, from 5.0 mg/kg oven dry pulp to 10000 mg/kg oven dry pulp, from 10.0 mg/kg oven dry pulp to 15000 mg/kg oven dry pulp, from 15.0 mg/kg oven dry pulp to 20000 mg/kg oven dry pulp, from 20.0 mg/kg oven dry pulp to 30000 mg/kg oven dry pulp, from 30.0 mg/kg oven dry pulp to 40000 mg/kg oven dry pulp, from 40.0 mg/kg oven dry pulp to 60000 mg/kg oven dry pulp, from 60.0 mg/kg oven dry pulp to 80000 mg/kg oven dry pulp, and from 80.0 mg/kg oven dry pulp to 100000 mg/kg oven dry pulp, or any combination of these intervals.

[0042] The concentration of the cellulase added in the present invention is from 0.05 mg/kg oven dry pulp to 100 mg/kg oven dry pulp such as a concentration selected from the group consisting of from 0.05 mg/kg oven dry pulp to 80.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 60.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 40.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 20.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 10.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 5.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 85.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 65.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 45.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 25.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 15.0 mg/kg oven dry pulp and from 0.50 mg/kg oven dry pulp to 5.0 mg/kg oven dry pulp, or any combination of these intervals.

[0043] The method according to the invention results in an improved viscosity control, thereby allowing the reduction in the production of dissolving pulp outside final viscosity specification/target, typically more than 50% (such as more than 60% or more than 70%) reduction in the production of off-grade dissolving pulp with respect to viscosity. In one embodiment the method according to the invention results in increased reactivity of the kraft and/or sulfite dissolving pulp, particularly the kraft dissolving pulp having an increased reactivity of at least 10% (such as at least 20% or at least 30%).

[0044] In a preferred embodiment, the method results in reduced viscosity and/or improved viscosity control in the dissolving pulp production process; and/or the method results in increased reactivity of the dissolving pulp, preferably increased Fock's reactivity related to the viscose making process and therefore allowing savings in CS.sub.2 and thus reducing costs and environmental impact; and/or the method results in increased content of oxidized groups of the dissolving pulp. This increase in oxidized groups can increase the reactivity of the dissolving pulp not only in terms of fiber swelling and chemical accessibility but also considering that more anchor points (carbonyl and/or carboxyl groups) in the cellulose will be available for subsequent derivatization processes in the production of cellulose derivatives.

[0045] In a preferred embodiment, the method of the present invention further comprises subjecting the dissolving pulp to a xylanase and/or a mannanase and/or a lipase and/or laccase and/or peroxidase.

[0046] A dissolving pulp made by the method described above is also part of the invention. A textile fiber or a derivatized cellulose made of the dissolving pulp described above is also part of the invention.

[0047] The invention also relates to use of a lytic polysaccharide monooxygenase for treatment of dissolving pulp.

[0048] Lytic Polysaccharide Monooxygenase (LPMO)

[0049] The term "lytic polysaccharide monooxygenase" means an enzyme that oxidizes sp(3) carbons in polysaccharides such as chitin, cellulose, and starch in the presence of an external electron donor and, as currently hypothesized, utilizes copper at the active site to activate molecular oxygen. At present those enzymes belong to Auxiliary Activity families AA9, AA10, AA11, AA13, AA14 and AA15 as defined in the database of carbohydrate active enzymes (http://www.cazy.org/).

[0050] In a first aspect, the LPMO comprises the following motifs:

TABLE-US-00001 [ILMV]-P-x(4,5)-G-x-Y-[ILMV]-x-R-x-[EQ]-x(4)-[HNQ] and [FW][TF]-K-[AIV],

[0051] wherein x is any amino acid, x(4,5) is any four or five contiguous amino acids, and x(4) is any four contiguous amino acids.

[0052] The LPMO comprising the above-noted motifs may further comprise:

TABLE-US-00002 H-x(1,2)-G-P-x(3)-[YW]-[AILMV], [EQ]-x-Y-x(2)-C-x-[EHQN]-[FILV]-x-[ILV], or H-x(1,2)-G-P-x(3)-[YW]-[AILMV] and [EQ]-x-Y-x(2)-C-x-[EHQN]-[FILV]-x-[ILV],

[0053] wherein xis any amino acid, x(1,2) is any one or two contiguous amino acids, x(3) is any three contiguous amino acids, and x(2) is any two contiguous amino acids.

[0054] In a preferred aspect, the LPMO further comprises H-x(1,2)-G-P-x(3)-[YW]-[AILMV]. In another preferred aspect, the LPMO further comprises [EQ]-x-Y-x(2)-C-x-[EHQN]-[FILV]-x-[ILV]. In another preferred aspect, the LPMO further comprises H-x(1,2)-G-P-x(3)-[YW]-[AILMV] and [EQ]-x-Y-x(2)-C-x-[EHQN]-[FILV]-x-[ILV].

[0055] In a second aspect, the LPMO comprises the following motif:

TABLE-US-00003 [ILMV]-P-x(4,5)-G-x-Y-[ILMV]-x-R-x-[EQ]-x(3)-A- [HNQ],

[0056] wherein x is any amino acid, x(4,5) is any 4 or 5 contiguous amino acids, and x(3) is any 3 contiguous amino acids. In the above motif, the accepted IUPAC single letter amino acid abbreviation is employed.

[0057] In one aspect, the LPMO comprises an amino acid sequence that has a sequence identity to the mature polypeptide of SEQ ID NO: 1 (Thielavia terrestris), SEQ ID NO: 2 (Lentinus similis), SEQ ID NO: 3 (Thermoascus aurantiacus), SEQ ID NO: 8 (Thielavia terrestris), SEQ ID NO: 9 (Thielavia terrestris), SEQ ID NO: 10 (Thielavia terrestris), SEQ ID NO: 11 (Thielavia terrestris), SEQ ID NO: 12 (Thielavia terrestris), SEQ ID NO: 13 (Thielavia terrestris), SEQ ID NO: 14 (Trichoderma reesei), SEQ ID NO: 15 (Myceliophthora thermophila), SEQ ID NO: 16 (Myceliophthora thermophila), SEQ ID NO: 17 (Myceliophthora thermophila), SEQ ID NO: 18 (Myceliophthora thermophila), SEQ ID NO: 19 (Myceliophthora thermophila), SEQ ID NO: 20 (Thermoascus aurantiacus), SEQ ID NO: 21 (Aspergillus fumigatus), SEQ ID NO: 22 (Penicillium pinophilum), SEQ ID NO: 23 (Thermoascus sp.), SEQ ID NO: 24 (Penicillium sp.), SEQ ID NO: 25 (Thielavia terrestris), SEQ ID NO: 26 (Thielavia terrestris), SEQ ID NO: 27 (Thielavia terrestris), SEQ ID NO: 28 (Thielavia terrestris), SEQ ID NO: 29 (Thielavia terrestris), SEQ ID NO: 30 (Thielavia terrestris), SEQ ID NO: 31 (Thielavia terrestris), SEQ ID NO: 32 (Thielavia terrestris), SEQ ID NO: 33 (Thielavia terrestris), SEQ ID NO: 34 (Thielavia terrestris), SEQ ID NO: 35 (Thielavia terrestris), SEQ ID NO: 36 (Thermoascus crustaceus), SEQ ID NO: 37 (Thermoascus crustaceus), or SEQ ID NO: 38 (Thermoascus crustaceus) of at least 50%, e.g., at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100%.

[0058] In another aspect, the LPMO is an artificial variant comprising a substitution, deletion, and/or insertion of one or more (or several) amino acids of the mature polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, or SEQ ID NO: 38; or a homologous sequence thereof, an allelic variant thereof, or a functional fragment thereof.

[0059] Preferably, amino acid changes are of a minor nature, that is conservative amino acid substitutions or insertions that do not significantly affect the folding and/or activity of the protein; small deletions, typically of one to about 30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue; a small linker peptide of up to about 20-25 residues; or a small extension that facilitates purification by changing net charge or another function, such as a poly-histidine tract, an antigenic epitope or a binding domain.

[0060] Examples of conservative substitutions are within the group of basic amino acids (arginine, lysine and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine and valine), aromatic amino acids (phenylalanine, tryptophan and tyrosine), and small amino acids (glycine, alanine, serine, threonine and methionine). Amino acid substitutions that do not generally alter specific activity are known in the art and are described, for example, by H. Neurath and R. L. Hill, 1979, In, The Proteins, Academic Press, New York. The most commonly occurring exchanges are Ala/Ser, Val/Ile, Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr, Ser/Asn, Ala/Val, Ser/Gly, Tyr/Phe, Ala/Pro, Lys/Arg, Asp/Asn, Leu/Ile, Leu/Val, Ala/Glu, and Asp/Gly.

[0061] Alternatively, the amino acid changes are of such a nature that the physico-chemical properties of the polypeptides are altered. For example, amino acid changes may improve the thermal stability of the polypeptide, alter the substrate specificity, change the pH optimum, and the like.

[0062] Essential amino acids in a parent polypeptide can be identified according to procedures known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham and Wells, 1989, Science 244: 1081-1085). In the latter technique, single alanine mutations are introduced at every residue in the molecule, and the resultant mutant molecules are tested for cellulolytic enhancing activity to identify amino acid residues that are critical to the activity of the molecule. See also, Hilton et al., 1996, J. Biol. Chem. 271: 4699-4708. The active site of the enzyme or other biological interaction can also be determined by physical analysis of structure, as determined by such techniques as nuclear magnetic resonance, crystallography, electron diffraction, or photoaffinity labeling, in conjunction with mutation of putative contact site amino acids. See, for example, de Vos et al., 1992, Science 255: 306-312; Smith et al., 1992, J. Mol. Biol. 224: 899-904; Wlodaver et al., 1992, FEBS Lett. 309: 59-64. The identities of essential amino acids can also be inferred from analysis of identities with polypeptides that are related to the parent polypeptide.

[0063] Single or multiple amino acid substitutions, deletions, and/or insertions can be made and tested using known methods of mutagenesis, recombination, and/or shuffling, followed by a relevant screening procedure, such as those disclosed by Reidhaar-Olson and Sauer, 1988, Science 241: 53-57; Bowie and Sauer, 1989, Proc. Natl. Acad. Sci. USA 86: 2152-2156; WO 95/17413; or WO 95/22625. Other methods that can be used include error-prone PCR, phage display (e.g., Lowman et al., 1991, Biochemistry 30: 10832-10837; U.S. Pat. No. 5,223,409; WO 92/06204), and region-directed mutagenesis (Derbyshire et al., 1986, Gene 46: 145; Ner et al., 1988, DNA 7: 127).

[0064] Mutagenesis/shuffling methods can be combined with high-throughput, automated screening methods to detect activity of cloned, mutagenized polypeptides expressed by host cells (Ness et al., 1999, Nature Biotechnology 17: 893-896). Mutagenized DNA molecules that encode active polypeptides can be recovered from the host cells and rapidly sequenced using standard methods in the art. These methods allow the rapid determination of the importance of individual amino acid residues in a polypeptide.

[0065] The total number of amino acid substitutions, deletions and/or insertions of the mature LPMO of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, or SEQ ID NO: 38 is not more than 10, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0066] In one aspect, the LPMO is used in the presence of a soluble activating divalent metal cation as described in WO 2008/151043, e.g., copper sulfate.

[0067] In one aspect, the LPMO is used in the presence of an electron donor thereof. The electron donor can exist in the dissolving pulp to be treated. In one embodiment, no or a little amount of electron donor can be added to the dissolving pulp. In another embodiment, an effective amount of electron donor can be added to the dissolving pulp.

[0068] In the present invention, the electron donor can be a dioxy compound, a bicylic compound, a heterocyclic compound, a nitrogen-containing compound, or a sulfur-containing compound.

[0069] The dioxy compound may include any suitable compound containing two or more oxygen atoms. In some aspects, the dioxy compounds contain a substituted aryl moiety as described herein. The dioxy compounds may comprise one or more (several) hydroxyl and/or hydroxyl derivatives, but also include substituted aryl moieties lacking hydroxyl and hydroxyl derivatives. Non-limiting examples of dioxy compounds include pyrocatechol or catechol; caffeic acid; 3,4-dihydroxybenzoic acid; 4-tert-butyl-5-methoxy-1,2-benzenediol; ascorbic acid, pyrogallol; gallic acid; methyl-3,4,5-trihydroxybenzoate; 2,3,4-trihydroxybenzophenone; 2,6-dimethoxyphenol; sinapinic acid; 3,5-dihydroxybenzoic acid; 4-chloro-1,2-benzenediol; 4-nitro-1,2-benzenediol; tannic acid; ethyl gallate; methyl glycolate; dihydroxyfumaric acid; 2-butyne-1,4-diol; (croconic acid; 1,3-propanediol; tartaric acid; 2,4-pentanediol; 3-ethyoxy-1,2-propanediol; 2,4,4'-trihydroxybenzophenone; cis-2-butene-1,4-diol; 3,4-dihydroxy-3-cyclobutene-1,2-dione; dihydroxyacetone; acrolein acetal; methyl-4-hydroxybenzoate; 4-hydroxybenzoic acid; and methyl-3,5-dimethoxy-4-hydroxybenzoate; or a salt or solvate thereof.

[0070] The bicyclic compound may include any suitable substituted fused ring system as described herein. The compounds may comprise one or more (several) additional rings, and are not limited to a specific number of rings unless otherwise stated. In one aspect, the bicyclic compound is a flavonoid. In another aspect, the bicyclic compound is an optionally subsituted isoflavonoid. In another aspect, the bicyclic compound is an optionally substituted flavylium ion, such as an optionally substituted anthocyanidin or optionally substituted anthocyanin, or derivative thereof. Non-limiting examples of bicyclic compounds include epicatechin; quercetin; myricetin; taxifolin; kaempferol; morin; acacetin; naringenin; isorhamnetin; apigenin; cyanidin; cyanin; kuromanin; keracyanin; or a salt or solvate thereof.

[0071] The heterocyclic compound may be any suitable compound, such as an optionally substituted aromatic or non-aromatic ring comprising a heteroatom, as described herein. In one aspect, the heterocyclic is a compound comprising an optionally substituted heterocycloalkyl moiety or an optionally substituted heteroaryl moiety. In another aspect, the optionally substituted heterocycloalkyl moiety or optionally substituted heteroaryl moiety is an optionally substituted 5-membered heterocycloalkyl or an optionally substituted 5-membered heteroaryl moiety. In another aspect, the optionally substituted heterocycloalkyl or optionally substituted heteroaryl moiety is an optionally substituted moiety selected from pyrazolyl, furanyl, imidazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrrolyl, pyridyl, pyrimidyl, pyridazinyl, thiazolyl, triazolyl, thienyl, dihydrothieno-pyrazolyl, thianaphthenyl, carbazolyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl, quinolinyl, benzotriazolyl, benzothiazolyl, benzooxazolyl, benzimidazolyl, isoquinolinyl, isoindolyl, acridinyl, benzoisazolyl, dimethylhydantoin, pyrazinyl, tetrahydrofuranyl, pyrrolinyl, pyrrolidinyl, morpholinyl, indolyl, diazepinyl, azepinyl, thiepinyl, piperidinyl, and oxepinyl. In another aspect, the optionally substituted heterocycloalkyl moiety or optionally substituted heteroaryl moiety is an optionally substituted furanyl. Non-limiting examples of heterocyclic compounds include (1,2-dihydroxyethyl)-3,4-dihydroxyfuran-2(5H)-one; 4-hydroxy-5-methyl-3-furanone; 5-hydroxy-2(5H)-furanone; [1,2-dihydroxyethyl]furan-2,3,4(5H)-trione; .alpha.-hydroxy-.gamma.-butyrolactone; ribonic .gamma.-lactone; aldohexuronicaldohexuronic acid .gamma.-lactone; gluconic acid .delta.-lactone; 4-hydroxycoumarin; dihydrobenzofuran; 5-(hydroxymethyl)furfural; furoin; 2(5H)-furanone; 5,6-dihydro-2H-pyran-2-one; and 5,6-dihydro-4-hydroxy-6-methyl-2H-pyran-2-one; or a salt or solvate thereof.

[0072] The nitrogen-containing compound may be any suitable compound with one or more nitrogen atoms. In one aspect, the nitrogen-containing compound comprises an amine, imine, hydroxylamine, or nitroxide moiety. Non-limiting examples of nitrogen-containing compounds include acetone oxime; violuric acid; pyridine-2-aldoxime; 2-aminophenol; 1,2-benzenediamine; 2,2,6,6-tetramethyl-1-piperidinyloxy; 5,6,7,8-tetrahydrobiopterin; 6,7-dimethyl-5,6,7,8-tetrahydropterine; and maleamic acid; or a salt or solvate thereof.

[0073] The quinone compound may be any suitable compound comprising a quinone moiety as described herein. Non-limiting examples of quinone compounds include 1,4-benzoquinone; 1,4-naphthoquinone; 2-hydroxy-1,4-naphthoquinone; 2,3-dimethoxy-5-methyl-1,4-benzoquinone or coenzyme Q.sub.0; 2,3,5,6-tetramethyl-1,4-benzoquinone or duroquinone; 1,4-dihydroxyanthraquinone; 3-hydroxy-1-methyl-5,6-indolinedione or adrenochrome; 4-tert-butyl-5-methoxy-1,2-benzoquinone; pyrroloquinoline quinone; or a salt or solvate thereof.

[0074] The sulfur-containing compound may be any suitable compound comprising one or more sulfur atoms. In one aspect, the sulfur-containing comprises a moiety selected from thionyl, thioether, sulfinyl, sulfonyl, sulfamide, sulfonamide, sulfonic acid, and sulfonic ester. Non-limiting examples of sulfur-containing compounds include ethanethiol; 2-propanethiol; 2-propene-1-thiol; 2-mercaptoethanesulfonic acid; benzenethiol; benzene-1,2-dithiol; cysteine; methionine; glutathione; cystine; or a salt or solvate thereof.

[0075] Cellulases

[0076] Cellulases or cellulolytic enzymes are enzymes involved in hydrolysis of cellulose. In the hydrolysis of native cellulose, it is known that there are three major types of cellulase enzymes involved, namely cellobiohydrolase (1,4-.beta.-D-glucan cellobiohydrolase, EC 3.2.1.91, e.g., cellobiohydrolase I and cellobiohydrolase II), endo-.beta.-1,4-glucanase (endo-1,4-.beta.-D-glucan 4-glucanohydrolase, EC 3.2.1.4) and .beta.-glucosidase (EC 3.2.1.21).

[0077] In order to be efficient, the digestion of cellulose and hemicellulose requires several types of enzymes acting cooperatively. At least three categories of enzymes are necessary to convert cellulose into fermentable sugars: endo-glucanases (EC 3.2.1.4) that cut the cellulose chains at random; cellobiohydrolases (EC 3.2.1.91) which cleave cellobiosyl units from the cellulose chain ends and beta-glucosidases (EC 3.2.1.21) that convert cellobiose and soluble cellodextrins into glucose. Among these three categories of enzymes involved in the biodegradation of cellulose, cellobiohydrolases are the key enzymes for the degradation of native crystalline cellulose. The term "cellobiohydrolase I" is defined herein as a cellulose 1,4-beta-cellobiosidase (also referred to as exo-glucanase, exo-cellobiohydrolase or 1,4-beta-cellobiohydrolase) activity, as defined in the enzyme class EC 3.2.1.91, which catalyzes the hydrolysis of 1,4-beta-D-glucosidic linkages in cellulose and cellotetraose, by the release of cellobiose from the non-reducing ends of the chains. The definition of the term "cellobiohydrolase II activity" is identical, except that cellobiohydrolase II attacks from the reducing ends of the chains.

[0078] Endoglucanases (EC No. 3.2.1.4) catalyses endo hydrolysis of 1,4-beta-D-glycosidic linkages in cellulose, cellulose derivatives (such as carboxy methyl cellulose and hydroxy ethyl cellulose), lichenin, beta-1,4 bonds in mixed beta-1,3 glucans such as cereal beta-D-glucans or xyloglucans and other plant material containing cellulosic parts. The authorized name is endo-1,4-beta-D-glucan 4-glucano hydrolase, but the abbreviated term endoglucanase is used in the present specification. In a preferred embodiment of the present invention, the cellulase used in the present invention is an endoglucanase.

[0079] The cellulases may comprise a carbohydrate-binding module (CBM) which enhances the binding of the enzyme to a cellulose-containing fiber and increases the efficacy of the catalytic active part of the enzyme. A CBM is defined as contiguous amino acid sequence within a carbohydrate-active enzyme with a discreet fold having carbohydrate-binding activity. For further information of CBMs see the CAZy internet server (Supra) or Tomme et al., (1995) in Enzymatic Degradation of Insoluble Polysaccharides (Saddler, J. N. & Penner, M., eds.), Cellulose-binding domains: classification and properties. pp. 142-163, American Chemical Society, Washington.

[0080] In a preferred embodiment the cellulases may be a preparation as defined in WO 2008/151079 , which is hereby incorporated by reference. The cellulase preparation may further comprise a beta-glucosidase, such as the fusion protein disclosed in US 60/832,511. In an embodiment the cellulase preparation also comprises a CBH II, preferably Thielavia terrestris cellobiohydrolase II CEL6A. In an embodiment the cellulase preparation also comprises a cellulase enzymes preparation, preferably the one derived from Trichoderma reesei. Cellulases are synthesized by a large number of microorganisms which include fungi, actinomycetes, myxobacteria and true bacteria but also by plants. Especially endoglucanases of a wide variety of specificities have been identified.

[0081] The cellulase activity may, in a preferred embodiment, be derived from a fungal source, such as a strain of the genus Trichoderma, preferably a strain of Trichoderma reesei; a strain of the genus Humicola, such as a strain of Humicola insolens; or a strain of Chrysosporium, preferably a strain of Chrysosporium lucknowense or a strain of the genus Thielavia, preferably Thielavia terrestris.

[0082] In one aspect, the cellulase comprises an amino acid sequence that has a sequence identity to SEQ ID NO: 4, the mature polypeptide of SEQ ID NO: 5, the mature polypeptide of SEQ ID NO: 6, or the mature polypeptide of SEQ ID NO: 7 of at least 50%, e.g., at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100%. In another aspect, the cellulase is an artificial variant comprising a substitution, deletion, and/or insertion of one or more (or several) amino acids of SEQ ID NO: 4, the mature polypeptide of SEQ ID NO: 5, the mature polypeptide of SEQ ID NO: 6, or the mature polypeptide of SEQ ID NO: 7; or a homologous sequence thereof, or an allelic variant thereof, or a functional fragment thereof. The total number of amino acid substitutions, deletions and/or insertions of SEQ ID NO: 4, the mature polypeptide of SEQ ID NO: 5, the mature polypeptide of SEQ ID NO: 6, or the mature polypeptide of SEQ ID NO: 7 is not more than 10, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0083] Fungi and bacteria produce a spectrum of cellulolytic enzymes (cellulases) which, on the basis of sequence similarities (hydrophobic cluster analysis), can be classified into different families of glycosyl hydrolases [Henrissat B & Bairoch A; Biochem. J. 1993 293 781-788]. At present are known cellulases belonging to the families 5, 6, 7, 8, 9, 10, 12, 26, 44, 45, 48, 60, and 61 of glycosyl hydrolases.

[0084] Additional Enzymes

[0085] Any enzyme having xylanase, mannanase, lipase, laccase, and/or peroxidase activity can be used as additional enzymes in the use and process of the invention. The additional enzymes and a lytic polysaccharide monooxygenase are added simultaneously or sequentially in any order. Below some non-limiting examples are listed of such additional enzymes. The enzymes written in capitals are commercial enzymes available from Novozymes A/S, Krogshoejvej 36, DK-2880 Bagsvaerd, Denmark. The activity of any of those additional enzymes can be analyzed using any method known in the art for the enzyme in question, including the methods mentioned in the references cited.

[0086] An example of a xylanase is the PULPZYME HC hemicellulase.

[0087] Examples of mannanases are the Trichoderma reesei endo-beta-mannanases described in Stahlbbrand et al, J. Biotechnol. 29 (1993), 229-242.

[0088] An example of a lipase is the RESINASE A2X lipase. An example of a xylanase is the PULPZYME HC hemicellulase.

[0089] Examples of peroxidases, and laccases are disclosed in EP 730641; WO 01/98469; EP 719337; EP 765394; EP 767836; EP 763115; and EP 788547. In the present context, whenever the peroxidase or laccase is mentioned that requires or benefits from the presence of acceptors (e.g., oxygen or hydrogen peroxide), enhancers, mediators and/or activators, such compounds should be considered to be included. Examples of enhancers and mediators are disclosed in EP 705327; WO 98/56899; EP 677102; EP 781328; and EP 707637. If desired a distinction could be made by defining a laccase or a peroxidase enzyme system as the combination of the enzyme in question and its acceptor, and optionally also an enhancer and/or mediator for the enzyme in question.

[0090] Temperature for the Method of the Present Invention

[0091] The temperature for the method of the present invention is typically from 20.degree. C. to 100.degree. C. such as a temperature interval selected from the group consisting of from 20.degree. C. to 30.degree. C., from 30.degree. C. to 40.degree. C., from 40.degree. C. to 50.degree. C., from 50.degree. C. to 60.degree. C., from 60.degree. C. to 70.degree. C., from 70.degree. C. to 80.degree. C., from 80.degree. C. to 90.degree. C., from 90.degree. C. to 100.degree. C., or any combination of these intervals.

[0092] Incubation Time for the Method of the Present Invention

[0093] The incubation time used for the mehod of the present invention is typically from 1 minute to 60 hours such as a time interval selected from the group consisting of from 1 minute to 15 minutes, from 15 minutes to 30 minutes, from 30 minutes to 45 minutes, from 45 minutes to 60 minutes, from 1 hour to 3 hours, from 3 hours to 6 hours, from 6 hours to 10 hours, from 10 hours to 12 hours, from 12 hours to 15 hours, from 15 hours to 20 hours, from 20 hours to 22 hours, from 22 hours to 25 hours, from 25 hours to 30 hours, from 30 hours to 40 hours, from 40 hours to 50 hours, from 50 hours to 60 hours, or any combination of these time intervals.

[0094] Pulp Used and Produced in the Method According to the Invention:

[0095] The dissolving pulp used in the present invention can be wood pulp coming e.g. from softwood trees (such as spruce, pine, fir, larch and hemlock) and/or hardwoods (such as eucalyptus, aspen and birch) or other plant sources such as bamboo.

[0096] In a preferred embodiment the dissolving pulp is selected from the group consisting of dissolving hardwood pulp and dissolving softwood pulp, or a mixture thereof.

[0097] The invention relates in one embodiment to dissolving pulp made by the method according to the invention. The invention relates in one embodiment to a kraft dissolving pulp or a sulfite dissolving pulp made by the method according to the invention.

[0098] The invention further relates to use of the dissolving pulp according to the invention for production of textile fibers. The dissolving pulp produced may be used in the manufacture of regenerated cellulose such as viscose, rayon, lyocell and modal fibers.

[0099] Performing the Method of the Invention in the Presence of One or More Surfactants

[0100] The method of the present invention can be performed in the presence of one or more surfactants such as one or more anionic surfactants and/or one or more nonionic surfactants and/or one or more cationic surfactants.

[0101] Surfactants can in one embodiment include poly(alkylene glycol)-based surfactants, ethoxylated dialkylphenols, ethoxylated dialkylphenols, ethoxylated alcohols and/or silicone based surfactants.

[0102] Examples of poly(alkylene glycol)-based surfactant are poly(ethylene glycol) alkyl ester, poly(ethylene glycol) alkyl ether, ethylene oxide/propylene oxide homo- and copolymers, or poly(ethylene oxide-co-propylene oxide) alkyl esters or ethers. Other examples include ethoxylated derivatives of primary alcohols, such as dodecanol, secondary alcohols, poly[propylene oxide], derivatives thereof, tridecylalcohol ethoxylated phosphate ester, and the like.

[0103] Specific presently preferred anionic surfactant materials useful in the practice of the invention comprise sodium alpha-sulfo methyl laurate, (which may include some alpha-sulfo ethyl laurate) for example as commercially available under the trade name ALPHA-STEP.TM.-ML40; sodium xylene sulfonate, for example as commercially available under the trade name STEPANATE.TM.-X; triethanolammonium lauryl sulfate, for example as commercially available under the trade name STEPANOL.TM.-WAT; diosodium lauryl sulfosuccinate, for example as commercially available under the trade name STEPAN.TM.-Mild SL3; further blends of various anionic surfactants may also be utilized, for example a 50%-50% or a 25%-75% blend of the aforesaid ALPHA-STEP.TM. and STEPANATE.TM. materials, or a 20%-80% blend of the aforesaid ALPHA-STEP.TM. and STEPANOL.TM. materials (all of the aforesaid commercially available materials may be obtained from Stepan Company, Northfield, Ill.).

[0104] Specific presently preferred nonionic surfactant materials useful in the practice of the invention comprise cocodiethanolamide, such as commercially available under trade name NINOL.TM.-11CM; alkyl polyoxyalkylene glycol ethers, such as relatively high molecular weight butyl ethylenoxide-propylenoxide block copolymers commercially available under the trade name TOXIMUL.TM.-8320 from the Stepan Company. Additional alkyl polyoxyalkylene glycol ethers may be selected, for example, as disclosed in U.S. Pat. No. 3,078,315. Blends of the various nonionic surfactants may also be utilized, for example a 50%-50% or a 25%-75% blend of the aforesaid NINOL.TM. and TOXIMUL.TM. materials.

[0105] Specific presently preferred anionic/nonionic surfactant blends useful in the practice of the invention include various mixtures of the above materials, for example a 50%-50% blends of the aforesaid ALPHA-STEP.TM. and NINOL.TM. materials or a 25%-75% blend of the aforesaid STEPANATE.TM. and TOXIMUL.TM. materials.

[0106] Preferably, the various anionic, nonionic and anionic/nonionic surfactant blends utilized in the practice of the invention have a solids or actives content up to about 100% by weight and preferably have an active content ranging from about 10% to about 80%. Of course, other blends or other solids (active) content may also be utilized and these anionic surfactants, nonionic surfactants, and mixtures thereof may also be utilized with known pulping chemicals such as, for example, anthraquinone and derivatives thereof and/or other typical paper chemicals, such as caustics, defoamers and the like.

[0107] The invention described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed, since these embodiments are intended as illustrations of several aspects of the invention. Any equivalent embodiments are intended to be within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. In the case of conflict, the present disclosure including definitions will control.

[0108] Various references are cited herein, the disclosures of which are incorporated by reference in their entireties.

EXAMPLES

[0109] Materials and Methods

[0110] The intrinsic viscosity of the pulp was measured according to ISO 5351 (International Organization for Standardization 5351).

[0111] The pulp viscosity was measured by mViPr according to WO 2011/107472 A9.

[0112] The amount of aldehyde groups (CHO content) was measured spectrophotometrically according to the procedure described by Obolenskaya et al., "Determination of aldehyde groups in oxidized pulps," Laboratory Manipulations in Wood and Cellulose Chemistry, Ecologia, Moscow, 211-212, 1991, which is based on the reaction of 2,3,5-triphenyltetrazolium chloride (TTC) with the aldehyde groups leading to the formation of formazan (red colorant).

[0113] The Fock's reactivity is a measure of how much of a known amount of pulp is reacted with CS.sub.2 as a small-scale simulation of the viscose making process and it was carried out at 9% NaOH.

TABLE-US-00004 Enzyme Description Thielavia Tt LPMO shown as the mature polypeptide of terrestris SEQ ID NO: 1 herein, also shown as the LPMO mature polypeptide of SEQ ID NO: 8 of (Tt LPMO) WO 2010/080532A Lentinus similis Ls LPMO shown as the mature polypeptide of LPMO SEQ ID NO: 2 herein, also shown as (Ls LPMO) the mature polypeptide of SEQ ID NO: 6 of WO2014/066141A Thermoascus Ta LPMO shown as the mature polypeptide of aurantiacus SEQ ID NO: 3 herein, also shown as the LPMO mature polypeptide of (Ta LPMO) SEQ ID NO: 2 of WO2005/074656A Endoglucanase-1 Endoglucanase shown as SEQ ID NO: 4 herein, which is a Q120H variant of the mature endoglucanase shown as SEQ ID NO: 9 of WO 96/29397A Endoglucanase-2 Endoglucanase shown as SEQ ID NO: 5 herein, which is a cellulase shown as the mature polypeptide of SEQ ID NO: 2 in WO1991/017243A Endoglucanase-3 Endoglucanase shown as SEQ ID NO: 6 herein, which is a GH45 cellulase shown as the mature polypeptide of SEQ ID NO: 2 in WO 2015/058700A Endoglucanase-4 Endoglucanase shown as SEQ ID NO: 7 herein, which is a GH5 endoglucanase shown as the mature polypeptide of SEQ ID NO: 2 in WO2013/019780A

Example 1

LPMO Treatment of Unbleached Hardwood Kraft Dissolving Pulp

[0114] Unbleached hardwood kraft dissolving pulp with a kappa number of 6.8 (TAPPI T 236 procedure), ISO brightness of 51% and an intrinsic viscosity of 1025 mL/g produced by a pre-hydrolysis kraft pulping process and further treated with a cold-caustic extraction stage from a dissolving pulp production process was used. This pulp was treated with several LPMOs in a small-scale assay using 24 mg of oven-dry fiber at 0.4% consistency, 45.degree. C., pH 5.0 (acetate buffer, 50 mM) for 20 hours. The enzyme treatment (denoted as X-stage) was done at a dosage of 5 mg EP (enzyme protein)/g odp (oven-dry pulp).

[0115] The pulp suspension at 0.4% consistency was disintegrated with a magnetic bar in glass test tubes placed in a heating block at 45.degree. C. Ascorbic acid, gallic acid or pyrogallol was added as electron donor to a final concentration of 1 mM in the suspension, followed by the addition of the LPMO enzyme to a final volume of 6 mL. After the 20 hour incubation time upon the addition of the enzyme, the tubes were cooled down in ice and then 6 mL of cupri-ethylenediamine (CED) solution added to dissolve the fibers. The pulp dissolution was done in a rotary agitator at a room temperature of 25.degree. C. for 25 min. Control experiments were done in the same way but without the addition of the enzyme.

[0116] After the dissolution time, the pulp viscosity was measured by mViPr. The mViPr pipette consists of a modified Gilson Concept C300 pipette equipped with a pressure sensor and Diamond D300 Gilson tips. Samples were kept at a constant temperature within .+-.0.1.degree. C. A volume of 200 .mu.L dissolved pulp was aspirated and dispensed in and out of the pipette, respectively, while recording the pressure in the pipette headspace. A pipette speed of 4 was applied. Apirations were followed by a 2s delay, and dispensing was followed by a 5s delay. Each sample measurement consisted of 15 aspiration-dispensing cycles, and pressure results were average of 15 aspiration or dispensing pressures, respectively.

[0117] The aspiration pressure results from the mViPr measurements are presented in Table 1. It can be seen that the LPMOs can reduce the average size of the cellulose which is expressed as the reduction in the solution viscosity as measured by the reduction in aspiration pressure. The performance of each LPMO is also dependent on the specific electron donor utilized. In addition, the electron donor itself also degrades cellulose when compared to the original pulp, particularly ascorbic acid. Tt LPMO is quite effective in reducing pulp viscosity with all electron donors.

TABLE-US-00005 TABLE 1 Aspiration pressure of the different unbleached pulps dissolved in CED Aspiration Reduction vs. Enzyme pressure (Pa) control (%) Original -1900.9 -- Control Ascorbic acid -1408.8 0 Ta LPMO + Ascorbic acid -1195.9 15 Ls LPMO + Ascorbic acid -1135.8 19 Tt LPMO + Ascorbic acid -656.6 53 Control Gallic acid -1753.8 0 Ta LPMO + Gallic acid -1089.5 38 Ls LPMO + Gallic acid -1091.6 38 Tt LPMO + Gallic acid -580.2 67 Control Pyrogallol -1605.9 0 Ta LPMO + Pyrogallol -923.7 42 Ls LPMO + Pyrogallol -232.9 85 Tt LPMO + Pyrogallol -655.9 59

Example 2

LPMO Treatment of Bleached Hardwood Kraft Dissolving Pulp

[0118] Bleached hardwood kraft never dried pulp of acetate-grade produced by a pre-hydrolysis kraft pulping process having an ISO brightness of 93.7% and an intrinsic viscosity of 684 mL/g was used. This pulp was treated with several LPMOs with three different electron donors using the same conditions and procedures as in Example 1.

[0119] The aspiration pressure results from the mViPr measurements are presented in Table 2 for the bleached dissolving pulps. LPMOs can reduce the average size of the cellulose molecules as expressed by the reduction in the solution (dissolved pulp) viscosity, measured by the reduction in aspiration pressure. Higher reduction in bleached pulp viscosity compared to controls (no enzyme) were achieved when the LPMOs were used together with gallic acic and pyrogallol as compared to the ascorbic acid which also reduces signfificantly viscosity itself. Tt LPMO is a more robust LPMO regarding viscosity reduction with all the electron donors used.

TABLE-US-00006 TABLE 2 Aspiration pressure of the different bleached pulps dissolved in CED Aspiration Reduction vs. Enzyme pressure (Pa) control (%) Original -450.9 -- Control Ascorbic acid -345.1 0 Ta LPMO + Ascorbic acid -339.5 2 Ls LPMO + Ascorbic acid -338.2 2 Tt LPMO + Ascorbic acid -244.5 29 Control Gallic acid -523.1 0 Ta LPMO + Gallic acid -245.3 53 Ls LPMO + Gallic acid -235.2 55 Tt LPMO + Gallic acid -160.3 69 Control Pyrogallol -433.7 0 Ta LPMO + Pyrogallol -260.9 40 Ls LPMO + Pyrogallol -88.8 80 Tt LPMO + Pyrogallol -43.7 90

Example 3

Effect of LPMO Treatment with/without Endoglucanase at Medium Pulp Consistency on Bleached Dissolving Pulp Viscosity, Reactivity and CHO Content

[0120] Bleached hardwood kraft dissolving pulp produced by a pre-hydrolysis kraft pulping process of viscose-grade was used having an intrinsic viscosity of 512 mL/g. This pulp was treated with several LPMOs using gallic acid (1 mM) as electron donor at 1.5% consistency in Distek vessels (Distek model Symphony 7100) with heating and continuous overhead stirring.

[0121] Once the pulps were disintegrated and at the temperature set-point of 45.degree. C., gallic acid was added and then the enzyme (LPMO and/or endoglucanase). The enzyme treatment of the pulp took 25.5 hour at pH 5.0 (acetate buffer, 50 mM) using 2 mg EP/g odp of LPMO and 1.2 mg EP/kg odp of endoglucanase. After the enzyme treatment, the pulps were filtered and washed in three consecutive steps with 1 L of tap water. Part of the pulp sample was dried before measuring the CHO content and Fock's reactivity and part of the sample was kept wet in the fridge to test for intrinsic viscosity.

[0122] In Table 3, it can be seen that with a lower dosage of 2 mg EP/g odp a small reduction in viscosity reduction is obtained with LPMOs tested, as compared to Examples 1 and 2 with a higher dosage of LPMOs. However, when combined with an endoglucanase, the viscosity reduction is enhanced with a surprising synergistic effect. The same synergy between LPMO and endoglucanase is seen in terms of Fock's reactivity and the amount of CHO groups.

TABLE-US-00007 TABLE 3 Intrinsic viscosity, Fock's reactivity and CHO content of the bleached dissolving pulps Intrinsic Fock's CHO viscosity reactivity content Enzyme (mL/g) (%) (mmol/kg odp) Original 512 16 17.8 Control Gallic acid 482 16 16.3 Tt LPMO + Gallic acid 479 20 19.8 Ta LPMO + Gallic acid 471 16 22.9 endoglucanase 480 15 19.7 Tt LPMO + endoglucanase + 429 26 27.7 Gallic acid

Example 4

Effect of Combined LPMO and Endoglucanase Treatment on Unbleached Hardwood Kraft Dissolving Pulp

[0123] The same unbleached hardwood kraft dissolving pulp of Example 1 was used and treated with several LPMOs using gallic acid (1 mM) as electron donor in combination with several endoglucanases in a small-scale assay using 24 mg of oven-dry fiber at 0.4% consistency, 50.degree. C., pH 6.0 (phosphate buffer, 50 mM) for 20 hours. The LPMO treatments were done at dosages of 2.5 mg EP (enzyme protein)/g odp (high dosage) or 0.5 mg EP/g odp (low dosage). The endoglucanase treatments were done at dosages of 0.5 mg EP/kg odp. The assay and viscosity measurements were performed using the same conditions and procedures as described in Example 1.

[0124] The aspiration pressure results from the mViPr measurements are presented in Table 4. It can be seen that the higher the dosage of LPMO, the higher the pulp viscosity reduction. A high dosage of LPMO gives a surprisingly higher viscosity reduction than the endoglucanase treatments. The treatment with a combination of LPMO and an endoglucanase shows better viscosity reduction performance than individual treatment with LPMO or endoglucanase. Using this pulp of high viscosity, it is observed a boosting effect on viscosity reduction by the use of the LPMO combined with the endoglucanase, which allows a significant reduction on the amount of LPMO needed. For instance, at a low Tt LPMO dosage (0.5 mg EP/g odp), the co-addition of an endoglucanase 2 and 3 gives even higher viscosity reduction than the high dosage of Tt LPMO alone (2.5 mg EP/g odp).

TABLE-US-00008 TABLE 4 Aspiration pressure of the different unbleached pulps dissolved in CED Aspiration Reduction pressure vs. Enzyme (Pa) control (%) Original -1594.1 -- Control Gallic acid -1565.3 0 Tt LPMO (high dose) + Gallic acid -821.0 48 Tt LPMO (low dose) + Gallic acid -1361.0 13 Ls LPMO (high dose) + Gallic acid -665.3 57 Ls LPMO (low dose) + Gallic acid -1141.0 27 Endoglucanase 1 -995.9 36 Tt LPMO (high dose) + Gallic acid + -415.1 73 endoglucanase 1 Tt LPMO (low dose) + Gallic acid + -883.2 44 endoglucanase 1 Ls LPMO (high dose) + Gallic acid + -646.2 59 endoglucanase 1 Ls LPMO (low dose) + Gallic acid + -905.4 42 endoglucanase 1 Endoglucanase 2 -1083.7 31 Tt LPMO (high dose) + Gallic acid + -341.9 78 endoglucanase 2 Tt LPMO (low dose) + Gallic acid + -762.6 51 endoglucanase 2 Ls LPMO (high dose) + Gallic acid + -485.7 69 endoglucanase 2 Ls LPMO (low dose) + Gallic acid + -969.1 38 endoglucanase 2 Endoglucanase 3 -1049.2 33 Tt LPMO (high dose) + Gallic acid + -441.7 72 endoglucanase 3 Tt LPMO (low dose) + Gallic acid + -506.1 68 endoglucanase 3 Ls LPMO (high dose) + Gallic acid + -284.6 82 endoglucanase 3 Ls LPMO (low dose) + Gallic acid + -315.3 80 endoglucanase 3 Endoglucanase 4 -1270.7 19 Tt LPMO (high dose) + Gallic acid + -732.9 53 endoglucanase 4 Tt LPMO (low dose) + Gallic acid + -1169.2 25 endoglucanase 4 Ls LPMO (high dose) + Gallic acid + -904.8 42 endoglucanase 4 Ls LPMO (low dose) + Gallic acid + -1186.5 24 endoglucanase 4

Sequence CWU 1

1

381226PRTThielavia terrestris 1Met Leu Ala Asn Gly Ala Ile Val Phe Leu Ala Ala Ala Leu Gly Val1 5 10 15Ser Gly His Tyr Thr Trp Pro Arg Val Asn Asp Gly Ala Asp Trp Gln 20 25 30Gln Val Arg Lys Ala Asp Asn Trp Gln Asp Asn Gly Tyr Val Gly Asp 35 40 45Val Thr Ser Pro Gln Ile Arg Cys Phe Gln Ala Thr Pro Ser Pro Ala 50 55 60Pro Ser Val Leu Asn Thr Thr Ala Gly Ser Thr Val Thr Tyr Trp Ala65 70 75 80Asn Pro Asp Val Tyr His Pro Gly Pro Val Gln Phe Tyr Met Ala Arg 85 90 95Val Pro Asp Gly Glu Asp Ile Asn Ser Trp Asn Gly Asp Gly Ala Val 100 105 110Trp Phe Lys Val Tyr Glu Asp His Pro Thr Phe Gly Ala Gln Leu Thr 115 120 125Trp Pro Ser Thr Gly Lys Ser Ser Phe Ala Val Pro Ile Pro Pro Cys 130 135 140Ile Lys Ser Gly Tyr Tyr Leu Leu Arg Ala Glu Gln Ile Gly Leu His145 150 155 160Val Ala Gln Ser Val Gly Gly Ala Gln Phe Tyr Ile Ser Cys Ala Gln 165 170 175Leu Ser Val Thr Gly Gly Gly Ser Thr Glu Pro Pro Asn Lys Val Ala 180 185 190Phe Pro Gly Ala Tyr Ser Ala Thr Asp Pro Gly Ile Leu Ile Asn Ile 195 200 205Tyr Tyr Pro Val Pro Thr Ser Tyr Gln Asn Pro Gly Pro Ala Val Phe 210 215 220Ser Cys2252254PRTLentinus similis 2Met Lys Tyr Ser Ile Leu Gly Leu Thr Ala Leu Ser Phe Val Ala Ser1 5 10 15Ala Ala Ala His Thr Leu Val Trp Gly Val Trp Val Asn Gly Val Asp 20 25 30Gln Gly Asp Gly Arg Asn Ile Tyr Ile Arg Ser Pro Pro Asn Asn Asn 35 40 45Pro Val Lys Asn Leu Thr Ser Pro Asp Met Thr Cys Asn Val Asp Asn 50 55 60Arg Val Val Pro Lys Ser Val Pro Val Asn Ala Gly Asp Thr Leu Thr65 70 75 80Phe Glu Trp Tyr His Asn Thr Arg Asp Asp Asp Ile Ile Ala Ser Ser 85 90 95His His Gly Pro Ile Ala Val Tyr Ile Ala Pro Ala Ala Ser Asn Gly 100 105 110Gln Gly Asn Val Trp Val Lys Leu Phe Glu Asp Ala Tyr Asn Val Thr 115 120 125Asn Ser Thr Trp Ala Val Asp Arg Leu Ile Thr Ala His Gly Gln His 130 135 140Ser Val Val Val Pro His Val Ala Pro Gly Asp Tyr Leu Phe Arg Ala145 150 155 160Glu Ile Ile Ala Leu His Glu Ala Asp Ser Leu Tyr Ser Gln Asn Pro 165 170 175Ile Arg Gly Ala Gln Phe Tyr Ile Ser Cys Ala Gln Ile Thr Ile Asn 180 185 190Ser Ser Asp Asp Ser Thr Pro Leu Pro Ala Gly Val Pro Phe Pro Gly 195 200 205Ala Tyr Thr Asp Ser Thr Pro Gly Ile Gln Phe Asn Ile Tyr Thr Thr 210 215 220Pro Ala Thr Ser Tyr Val Ala Pro Pro Pro Ser Val Trp Ser Gly Ala225 230 235 240Leu Gly Gly Ser Ile Ala Gln Val Gly Asp Ala Ser Leu Glu 245 2503249PRTThermoascus aurantiacus 3Met Ser Phe Ser Lys Ile Ile Ala Thr Ala Gly Val Leu Ala Ser Ala1 5 10 15Ser Leu Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp Gly 20 25 30Lys Asn Tyr Gly Gly Tyr Leu Val Asn Gln Tyr Pro Tyr Met Ser Asn 35 40 45Pro Pro Glu Val Ile Ala Trp Ser Thr Thr Ala Thr Asp Leu Gly Phe 50 55 60Val Asp Gly Thr Gly Tyr Gln Thr Pro Asp Ile Ile Cys His Arg Gly65 70 75 80Ala Lys Pro Gly Ala Leu Thr Ala Pro Val Ser Pro Gly Gly Thr Val 85 90 95Glu Leu Gln Trp Thr Pro Trp Pro Asp Ser His His Gly Pro Val Ile 100 105 110Asn Tyr Leu Ala Pro Cys Asn Gly Asp Cys Ser Thr Val Asp Lys Thr 115 120 125Gln Leu Glu Phe Phe Lys Ile Ala Glu Ser Gly Leu Ile Asn Asp Asp 130 135 140Asn Pro Pro Gly Ile Trp Ala Ser Asp Asn Leu Ile Ala Ala Asn Asn145 150 155 160Ser Trp Thr Val Thr Ile Pro Thr Thr Ile Ala Pro Gly Asn Tyr Val 165 170 175Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gln Asn Gln Asp Gly 180 185 190Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Gln Val Thr Gly Gly Gly 195 200 205Ser Asp Asn Pro Ala Gly Thr Leu Gly Thr Ala Leu Tyr His Asp Thr 210 215 220Asp Pro Gly Ile Leu Ile Asn Ile Tyr Gln Lys Leu Ser Ser Tyr Ile225 230 235 240Ile Pro Gly Pro Pro Leu Tyr Thr Gly 2454278PRTArtificialQ120H variant of the mature endoglucanase shown as SEQ ID NO 9 of WO 96/29397 4Ala Ser Gly Ser Gly Gln Ser Thr Arg Tyr Trp Asp Cys Cys Lys Pro1 5 10 15Ser Cys Ala Trp Pro Gly Lys Ala Ala Val Ser Gln Pro Val Tyr Ala 20 25 30Cys Asp Ala Asn Phe Gln Arg Leu Ser Asp Phe Asn Val Gln Ser Gly 35 40 45Cys Asn Gly Gly Ser Ala Tyr Ser Cys Ala Asp Gln Thr Pro Trp Ala 50 55 60Val Asn Asp Asn Leu Ala Tyr Gly Phe Ala Ala Thr Ser Ile Ala Gly65 70 75 80Gly Ser Glu Ser Ser Trp Cys Cys Ala Cys Tyr Ala Leu Thr Phe Thr 85 90 95Ser Gly Pro Val Ala Gly Lys Thr Met Val Val Gln Ser Thr Ser Thr 100 105 110Gly Gly Asp Leu Gly Ser Asn His Phe Asp Ile Ala Met Pro Gly Gly 115 120 125Gly Val Gly Ile Phe Asn Gly Cys Ser Ser Gln Phe Gly Gly Leu Pro 130 135 140Gly Ala Gln Tyr Gly Gly Ile Ser Ser Arg Asp Gln Cys Asp Ser Phe145 150 155 160Pro Ala Pro Leu Lys Pro Gly Cys Gln Trp Arg Phe Asp Trp Phe Gln 165 170 175Asn Ala Asp Asn Pro Thr Phe Thr Phe Gln Gln Val Gln Cys Pro Ala 180 185 190Glu Ile Val Ala Arg Ser Gly Cys Lys Arg Asn Asp Asp Ser Ser Phe 195 200 205Pro Val Phe Thr Pro Pro Ser Gly Gly Asn Gly Gly Thr Gly Thr Pro 210 215 220Thr Ser Thr Ala Pro Gly Ser Gly Gln Thr Ser Pro Gly Gly Gly Ser225 230 235 240Gly Cys Thr Ser Gln Lys Trp Ala Gln Cys Gly Gly Ile Gly Phe Ser 245 250 255Gly Cys Thr Thr Cys Val Ser Gly Thr Thr Cys Gln Lys Leu Asn Asp 260 265 270Tyr Tyr Ser Gln Cys Leu 2755305PRTHumicola insolens 5Met Arg Ser Ser Pro Leu Leu Pro Ser Ala Val Val Ala Ala Leu Pro1 5 10 15Val Leu Ala Leu Ala Ala Asp Gly Arg Ser Thr Arg Tyr Trp Asp Cys 20 25 30Cys Lys Pro Ser Cys Gly Trp Ala Lys Lys Ala Pro Val Asn Gln Pro 35 40 45Val Phe Ser Cys Asn Ala Asn Phe Gln Arg Ile Thr Asp Phe Asp Ala 50 55 60Lys Ser Gly Cys Glu Pro Gly Gly Val Ala Tyr Ser Cys Ala Asp Gln65 70 75 80Thr Pro Trp Ala Val Asn Asp Asp Phe Ala Leu Gly Phe Ala Ala Thr 85 90 95Ser Ile Ala Gly Ser Asn Glu Ala Gly Trp Cys Cys Ala Cys Tyr Glu 100 105 110Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Lys Met Val Val Gln 115 120 125Ser Thr Ser Thr Gly Gly Asp Leu Gly Ser Asn His Phe Asp Leu Asn 130 135 140Ile Pro Gly Gly Gly Val Gly Ile Phe Asp Gly Cys Thr Pro Gln Phe145 150 155 160Gly Gly Leu Pro Gly Gln Arg Tyr Gly Gly Ile Ser Ser Arg Asn Glu 165 170 175Cys Asp Arg Phe Pro Asp Ala Leu Lys Pro Gly Cys Tyr Trp Arg Phe 180 185 190Asp Trp Phe Lys Asn Ala Asp Asn Pro Ser Phe Ser Phe Arg Gln Val 195 200 205Gln Cys Pro Ala Glu Leu Val Ala Arg Thr Gly Cys Arg Arg Asn Asp 210 215 220Asp Gly Asn Phe Pro Ala Val Gln Ile Pro Ser Ser Ser Thr Ser Ser225 230 235 240Pro Val Asn Gln Pro Thr Ser Thr Ser Thr Thr Ser Thr Ser Thr Thr 245 250 255Ser Ser Pro Pro Val Gln Pro Thr Thr Pro Ser Gly Cys Thr Ala Glu 260 265 270Arg Trp Ala Gln Cys Gly Gly Asn Gly Trp Ser Gly Cys Thr Thr Cys 275 280 285Val Ala Gly Ser Thr Cys Thr Lys Ile Asn Asp Trp Tyr His Gln Cys 290 295 300Leu3056293PRTNeurospora tetrasperma 6Met Arg Ser Ser Thr Val Leu Gln Thr Gly Leu Val Ala Ala Leu Pro1 5 10 15Phe Ala Val Gln Ala Ala Ser Gly Ser Gly Gln Ser Thr Arg Tyr Trp 20 25 30Asp Cys Cys Lys Pro Ser Cys Ser Trp Ser Gly Lys Ala Ser Val Asn 35 40 45Arg Pro Val Leu Ala Cys Asp Ala Asn Asn Asn Pro Leu Ser Asp Ala 50 55 60Ser Val Lys Ser Gly Cys Asp Gly Gly Ser Ala Tyr Thr Cys Ala Asn65 70 75 80Asn Ser Pro Trp Ala Val Asn Asp Gln Leu Ser Tyr Gly Phe Ala Ala 85 90 95Thr Lys Leu Ser Gly Gly Thr Glu Ser Ser Trp Cys Cys Ala Cys Tyr 100 105 110Ala Leu Thr Phe Thr Ser Gly Pro Val Ala Gly Lys Thr Met Val Val 115 120 125Gln Ser Thr Ser Thr Gly Gly Asp Leu Gly Ser Asn His Phe Asp Ile 130 135 140Asn Met Pro Gly Gly Gly Val Gly Leu Phe Asp Gly Cys Thr Arg Gln145 150 155 160Phe Gly Gly Leu Pro Gly Ala Gln Tyr Gly Gly Ile Ser Ser Arg Ser 165 170 175Gln Cys Asp Ser Phe Pro Ala Ala Leu Lys Pro Gly Cys Gln Trp Arg 180 185 190Phe Asp Trp Phe Gln Asn Ala Asp Asn Pro Asn Phe Thr Phe Lys Gln 195 200 205Val Gln Cys Pro Ser Glu Leu Thr Ser Arg Thr Gly Cys Lys Arg Asn 210 215 220Asp Asp Ser Gln Phe Pro Val Phe Thr Pro Pro Ser Gly Gly Gly Thr225 230 235 240Asn Pro Ser Thr Pro Thr Thr Pro Pro Ser Ser Gly Gly Gly Ser Gly 245 250 255Cys Thr Ala Asp Lys Tyr Ala Gln Cys Gly Gly Ser Gly Trp Ser Gly 260 265 270Cys Thr Asn Cys Pro Ser Gly Ser Thr Cys Lys Thr Ile Asn Asp Tyr 275 280 285Tyr His Gln Cys Ala 2907409PRTTalaromyces leycettanus 7Met Lys Phe Ser Asn Val Ile Leu Ala Ala Ser Ala Ser Ser Leu Val1 5 10 15Leu Ala Ala Pro Lys Ser Lys Thr Lys Arg Thr Ser Ala Phe Gln Trp 20 25 30Phe Gly Ala Asn Glu Ser Gly Ala Glu Phe Gly Asn Gln Asn Ile Pro 35 40 45Gly Thr Leu Gly Thr Asp Tyr Thr Trp Pro Asp Thr Ser Thr Ile Gln 50 55 60Thr Leu Arg Asn Ala Gly Met Asn Ile Phe Arg Val Pro Phe Leu Met65 70 75 80Glu Arg Leu Val Pro Asn Gln Met Thr Gly Ser Pro Asp Pro Thr Tyr 85 90 95Leu Ala Asp Leu Lys Ser Thr Val Asn Phe Ile Thr Gly Thr Gly Ala 100 105 110Tyr Ala Val Val Asp Pro His Asn Tyr Gly Arg Tyr Tyr Asn Asn Ile 115 120 125Ile Thr Ser Thr Ser Asp Phe Ala Ala Phe Trp Thr Thr Val Ala Ser 130 135 140Gln Phe Ala Ser Asn Pro Arg Val Ile Phe Asp Thr Asn Asn Glu Tyr145 150 155 160Asn Asn Met Asp Gln Thr Leu Val Leu Asn Leu Asn Gln Ala Ala Ile 165 170 175Asn Ala Ile Arg Ala Ala Gly Ala Thr Ser Gln Tyr Ile Phe Ala Glu 180 185 190Gly Asn Ser Trp Thr Gly Ala Trp Thr Trp Thr Ser Val Asn Asp Asn 195 200 205Met Lys Gln Leu Thr Asp Pro Ser Asn Lys Leu Val Tyr Glu Met His 210 215 220Gln Tyr Leu Asp Ser Asp Gly Ser Gly Thr Ser Asp Gln Cys Val Asn225 230 235 240Ser Thr Ile Gly Tyr Asp Arg Ile Val Ser Ala Thr Gln Trp Leu Gln 245 250 255Ala Asn Gly Lys Val Ala Phe Leu Gly Glu Phe Ala Gly Gly Ser Asn 260 265 270Ser Val Cys Glu Ala Ala Val Thr Gly Met Leu Asp Tyr Met Glu Gln 275 280 285Asn Ser Asp Val Trp Leu Gly Ala Glu Trp Trp Ala Ala Gly Pro Trp 290 295 300Trp Gly Asn Tyr Ile Tyr Ser Met Glu Pro Pro Ser Gly Ile Ala Tyr305 310 315 320Gln Asn Tyr Leu Ser Ile Leu Glu Pro Tyr Phe Pro Gly Gly Ser Tyr 325 330 335Ser Gly Gly Thr Gly Ser Gly Ser Gly Ser Thr Thr Thr Thr Ala Thr 340 345 350Thr Thr Thr Thr Lys Val Pro Pro Thr Ser Thr Thr Ser Ser Ala Ser 355 360 365Ser Thr Gly Thr Gly Val Ala Gln His Trp Gly Gln Cys Gly Gly Gln 370 375 380Gly Trp Thr Gly Pro Thr Thr Cys Val Ser Pro Tyr Thr Cys Gln Glu385 390 395 400Leu Asn Pro Tyr Tyr Tyr Gln Cys Leu 4058326PRTThielavia terrestris 8Met Lys Ser Phe Thr Ile Ala Ala Leu Ala Ala Leu Trp Ala Gln Glu1 5 10 15Ala Ala Ala His Ala Thr Phe Gln Asp Leu Trp Ile Asp Gly Val Asp 20 25 30Tyr Gly Ser Gln Cys Val Arg Leu Pro Ala Ser Asn Ser Pro Val Thr 35 40 45Asn Val Ala Ser Asp Asp Ile Arg Cys Asn Val Gly Thr Ser Arg Pro 50 55 60Thr Val Lys Cys Pro Val Lys Ala Gly Ser Thr Val Thr Ile Glu Met65 70 75 80His Gln Gln Pro Gly Asp Arg Ser Cys Ala Asn Glu Ala Ile Gly Gly 85 90 95Asp His Tyr Gly Pro Val Met Val Tyr Met Ser Lys Val Asp Asp Ala 100 105 110Val Thr Ala Asp Gly Ser Ser Gly Trp Phe Lys Val Phe Gln Asp Ser 115 120 125Trp Ala Lys Asn Pro Ser Gly Ser Thr Gly Asp Asp Asp Tyr Trp Gly 130 135 140Thr Lys Asp Leu Asn Ser Cys Cys Gly Lys Met Asn Val Lys Ile Pro145 150 155 160Glu Asp Ile Glu Pro Gly Asp Tyr Leu Leu Arg Ala Glu Val Ile Ala 165 170 175Leu His Val Ala Ala Ser Ser Gly Gly Ala Gln Phe Tyr Met Ser Cys 180 185 190Tyr Gln Leu Thr Val Thr Gly Ser Gly Ser Ala Thr Pro Ser Thr Val 195 200 205Asn Phe Pro Gly Ala Tyr Ser Ala Ser Asp Pro Gly Ile Leu Ile Asn 210 215 220Ile His Ala Pro Met Ser Thr Tyr Val Val Pro Gly Pro Thr Val Tyr225 230 235 240Ala Gly Gly Ser Thr Lys Ser Ala Gly Ser Ser Cys Ser Gly Cys Glu 245 250 255Ala Thr Cys Thr Val Gly Ser Gly Pro Ser Ala Thr Leu Thr Gln Pro 260 265 270Thr Ser Thr Ala Thr Ala Thr Ser Ala Pro Gly Gly Gly Gly Ser Gly 275 280 285Cys Thr Ala Ala Lys Tyr Gln Gln Cys Gly Gly Thr Gly Tyr Thr Gly 290 295 300Cys Thr Thr Cys Ala Ser Gly Ser Thr Cys Ser Ala Val Ser Pro Pro305 310 315 320Tyr Tyr Ser Gln Cys Leu 3259239PRTThielavia terrestris 9Met Arg Phe Asp Ala Leu Ser Ala Leu Ala Leu Ala Pro Leu Val Ala1 5 10 15Gly His Gly Ala Val Thr Ser Tyr Ile Ile Gly Gly Lys Thr Tyr Pro 20 25 30Gly Tyr Glu Gly Phe Ser Pro Ala Ser Ser Pro Pro Thr Ile Gln Tyr 35 40 45Gln Trp Pro Asp Tyr Asn Pro Thr Leu Ser Val Thr Asp Pro Lys Met 50 55 60Arg Cys Asn Gly Gly Thr Ser Ala Glu Leu Ser Ala Pro Val Gln Ala65 70 75 80Gly Glu Asn Val Thr Ala Val Trp Lys Gln Trp Thr His Gln Gln Gly 85 90 95Pro Val Met Val Trp Met Phe

Lys Cys Pro Gly Asp Phe Ser Ser Ser 100 105 110His Gly Asp Gly Lys Gly Trp Phe Lys Ile Asp Gln Leu Gly Leu Trp 115 120 125Gly Asn Asn Leu Asn Ser Asn Asn Trp Gly Thr Ala Ile Val Tyr Lys 130 135 140Thr Leu Gln Trp Ser Asn Pro Ile Pro Lys Asn Leu Ala Pro Gly Asn145 150 155 160Tyr Leu Ile Arg His Glu Leu Leu Ala Leu His Gln Ala Asn Thr Pro 165 170 175Gln Phe Tyr Ala Glu Cys Ala Gln Leu Val Val Ser Gly Ser Gly Ser 180 185 190Ala Leu Pro Pro Ser Asp Tyr Leu Tyr Ser Ile Pro Val Tyr Ala Pro 195 200 205Gln Asn Asp Pro Gly Ile Thr Val Asp Ile Tyr Asn Gly Gly Leu Thr 210 215 220Ser Tyr Thr Pro Pro Gly Gly Pro Val Trp Ser Gly Phe Glu Phe225 230 23510258PRTThielavia terrestris 10Met Leu Leu Thr Ser Val Leu Gly Ser Ala Ala Leu Leu Ala Ser Gly1 5 10 15Ala Ala Ala His Gly Ala Val Thr Ser Tyr Ile Ile Ala Gly Lys Asn 20 25 30Tyr Pro Gly Tyr Gln Gly Phe Ser Pro Ala Asn Ser Pro Asn Val Ile 35 40 45Gln Trp Gln Trp His Asp Tyr Asn Pro Val Leu Ser Cys Ser Asp Ser 50 55 60Lys Leu Arg Cys Asn Gly Gly Thr Ser Ala Thr Leu Asn Ala Thr Ala65 70 75 80Ala Pro Gly Asp Thr Ile Thr Ala Ile Trp Ala Gln Trp Thr His Ser 85 90 95Gln Gly Pro Ile Leu Val Trp Met Tyr Lys Cys Pro Gly Ser Phe Ser 100 105 110Ser Cys Asp Gly Ser Gly Ala Gly Trp Phe Lys Ile Asp Glu Ala Gly 115 120 125Phe His Gly Asp Gly Val Lys Val Phe Leu Asp Thr Glu Asn Pro Ser 130 135 140Gly Trp Asp Ile Ala Lys Leu Val Gly Gly Asn Lys Gln Trp Ser Ser145 150 155 160Lys Val Pro Glu Gly Leu Ala Pro Gly Asn Tyr Leu Val Arg His Glu 165 170 175Leu Ile Ala Leu His Gln Ala Asn Asn Pro Gln Phe Tyr Pro Glu Cys 180 185 190Ala Gln Val Val Ile Thr Gly Ser Gly Thr Ala Gln Pro Asp Ala Ser 195 200 205Tyr Lys Ala Ala Ile Pro Gly Tyr Cys Asn Gln Asn Asp Pro Asn Ile 210 215 220Lys Val Pro Ile Asn Asp His Ser Ile Pro Gln Thr Tyr Lys Ile Pro225 230 235 240Gly Pro Pro Val Phe Lys Gly Thr Ala Ser Lys Lys Ala Arg Asp Phe 245 250 255Thr Ala11226PRTThielavia terrestris 11Met Leu Ala Asn Gly Ala Ile Val Phe Leu Ala Ala Ala Leu Gly Val1 5 10 15Ser Gly His Tyr Thr Trp Pro Arg Val Asn Asp Gly Ala Asp Trp Gln 20 25 30Gln Val Arg Lys Ala Asp Asn Trp Gln Asp Asn Gly Tyr Val Gly Asp 35 40 45Val Thr Ser Pro Gln Ile Arg Cys Phe Gln Ala Thr Pro Ser Pro Ala 50 55 60Pro Ser Val Leu Asn Thr Thr Ala Gly Ser Thr Val Thr Tyr Trp Ala65 70 75 80Asn Pro Asp Val Tyr His Pro Gly Pro Val Gln Phe Tyr Met Ala Arg 85 90 95Val Pro Asp Gly Glu Asp Ile Asn Ser Trp Asn Gly Asp Gly Ala Val 100 105 110Trp Phe Lys Val Tyr Glu Asp His Pro Thr Phe Gly Ala Gln Leu Thr 115 120 125Trp Pro Ser Thr Gly Lys Ser Ser Phe Ala Val Pro Ile Pro Pro Cys 130 135 140Ile Lys Ser Gly Tyr Tyr Leu Leu Arg Ala Glu Gln Ile Gly Leu His145 150 155 160Val Ala Gln Ser Val Gly Gly Ala Gln Phe Tyr Ile Ser Cys Ala Gln 165 170 175Leu Ser Val Thr Gly Gly Gly Ser Thr Glu Pro Pro Asn Lys Val Ala 180 185 190Phe Pro Gly Ala Tyr Ser Ala Thr Asp Pro Gly Ile Leu Ile Asn Ile 195 200 205Tyr Tyr Pro Val Pro Thr Ser Tyr Gln Asn Pro Gly Pro Ala Val Phe 210 215 220Ser Cys22512304PRTThielavia terrestris 12Met Lys Gly Leu Phe Ser Ala Ala Ala Leu Ser Leu Ala Val Gly Gln1 5 10 15Ala Ser Ala His Tyr Ile Phe Gln Gln Leu Ser Ile Asn Gly Asn Gln 20 25 30Phe Pro Val Tyr Gln Tyr Ile Arg Lys Asn Thr Asn Tyr Asn Ser Pro 35 40 45Val Thr Asp Leu Thr Ser Asp Asp Leu Arg Cys Asn Val Gly Ala Gln 50 55 60Gly Ala Gly Thr Asp Thr Val Thr Val Lys Ala Gly Asp Gln Phe Thr65 70 75 80Phe Thr Leu Asp Thr Pro Val Tyr His Gln Gly Pro Ile Ser Ile Tyr 85 90 95Met Ser Lys Ala Pro Gly Ala Ala Ser Asp Tyr Asp Gly Ser Gly Gly 100 105 110Trp Phe Lys Ile Lys Asp Trp Gly Pro Thr Phe Asn Ala Asp Gly Thr 115 120 125Ala Thr Trp Asp Met Ala Gly Ser Tyr Thr Tyr Asn Ile Pro Thr Cys 130 135 140Ile Pro Asp Gly Asp Tyr Leu Leu Arg Ile Gln Ser Leu Ala Ile His145 150 155 160Asn Pro Trp Pro Ala Gly Ile Pro Gln Phe Tyr Ile Ser Cys Ala Gln 165 170 175Ile Thr Val Thr Gly Gly Gly Asn Gly Asn Pro Gly Pro Thr Ala Leu 180 185 190Ile Pro Gly Ala Phe Lys Asp Thr Asp Pro Gly Tyr Thr Val Asn Ile 195 200 205Tyr Thr Asn Phe His Asn Tyr Thr Val Pro Gly Pro Glu Val Phe Ser 210 215 220Cys Asn Gly Gly Gly Ser Asn Pro Pro Pro Pro Val Ser Ser Ser Thr225 230 235 240Pro Ala Thr Thr Thr Leu Val Thr Ser Thr Arg Thr Thr Ser Ser Thr 245 250 255Ser Ser Ala Ser Thr Pro Ala Ser Thr Gly Gly Cys Thr Val Ala Lys 260 265 270Trp Gly Gln Cys Gly Gly Asn Gly Tyr Thr Gly Cys Thr Thr Cys Ala 275 280 285Ala Gly Ser Thr Cys Ser Lys Gln Asn Asp Tyr Tyr Ser Gln Cys Leu 290 295 30013317PRTThielavia terrestris 13Met Lys Gly Leu Ser Leu Leu Ala Ala Ala Ser Ala Ala Thr Ala His1 5 10 15Thr Ile Phe Val Gln Leu Glu Ser Gly Gly Thr Thr Tyr Pro Val Ser 20 25 30Tyr Gly Ile Arg Asp Pro Ser Tyr Asp Gly Pro Ile Thr Asp Val Thr 35 40 45Ser Asp Ser Leu Ala Cys Asn Gly Pro Pro Asn Pro Thr Thr Pro Ser 50 55 60Pro Tyr Ile Ile Asn Val Thr Ala Gly Thr Thr Val Ala Ala Ile Trp65 70 75 80Arg His Thr Leu Thr Ser Gly Pro Asp Asp Val Met Asp Ala Ser His 85 90 95Lys Gly Pro Thr Leu Ala Tyr Leu Lys Lys Val Asp Asp Ala Leu Thr 100 105 110Asp Thr Gly Ile Gly Gly Gly Trp Phe Lys Ile Gln Glu Ala Gly Tyr 115 120 125Asp Asn Gly Asn Trp Ala Thr Ser Thr Val Ile Thr Asn Gly Gly Phe 130 135 140Gln Tyr Ile Asp Ile Pro Ala Cys Ile Pro Asn Gly Gln Tyr Leu Leu145 150 155 160Arg Ala Glu Met Ile Ala Leu His Ala Ala Ser Thr Gln Gly Gly Ala 165 170 175Gln Leu Tyr Met Glu Cys Ala Gln Ile Asn Val Val Gly Gly Ser Gly 180 185 190Ser Ala Ser Pro Gln Thr Tyr Ser Ile Pro Gly Ile Tyr Gln Ala Thr 195 200 205Asp Pro Gly Leu Leu Ile Asn Ile Tyr Ser Met Thr Pro Ser Ser Gln 210 215 220Tyr Thr Ile Pro Gly Pro Pro Leu Phe Thr Cys Ser Gly Ser Gly Asn225 230 235 240Asn Gly Gly Gly Ser Asn Pro Ser Gly Gly Gln Thr Thr Thr Ala Lys 245 250 255Pro Thr Thr Thr Thr Ala Ala Thr Thr Thr Ser Ser Ala Ala Pro Thr 260 265 270Ser Ser Gln Gly Gly Ser Ser Gly Cys Thr Val Pro Gln Trp Gln Gln 275 280 285Cys Gly Gly Ile Ser Phe Thr Gly Cys Thr Thr Cys Ala Ala Gly Tyr 290 295 300Thr Cys Lys Tyr Leu Asn Asp Tyr Tyr Ser Gln Cys Gln305 310 31514249PRTTrichoderma reesei 14Met Lys Ser Cys Ala Ile Leu Ala Ala Leu Gly Cys Leu Ala Gly Ser1 5 10 15Val Leu Gly His Gly Gln Val Gln Asn Phe Thr Ile Asn Gly Gln Tyr 20 25 30Asn Gln Gly Phe Ile Leu Asp Tyr Tyr Tyr Gln Lys Gln Asn Thr Gly 35 40 45His Phe Pro Asn Val Ala Gly Trp Tyr Ala Glu Asp Leu Asp Leu Gly 50 55 60Phe Ile Ser Pro Asp Gln Tyr Thr Thr Pro Asp Ile Val Cys His Lys65 70 75 80Asn Ala Ala Pro Gly Ala Ile Ser Ala Thr Ala Ala Ala Gly Ser Asn 85 90 95Ile Val Phe Gln Trp Gly Pro Gly Val Trp Pro His Pro Tyr Gly Pro 100 105 110Ile Val Thr Tyr Val Val Glu Cys Ser Gly Ser Cys Thr Thr Val Asn 115 120 125Lys Asn Asn Leu Arg Trp Val Lys Ile Gln Glu Ala Gly Ile Asn Tyr 130 135 140Asn Thr Gln Val Trp Ala Gln Gln Asp Leu Ile Asn Gln Gly Asn Lys145 150 155 160Trp Thr Val Lys Ile Pro Ser Ser Leu Arg Pro Gly Asn Tyr Val Phe 165 170 175Arg His Glu Leu Leu Ala Ala His Gly Ala Ser Ser Ala Asn Gly Met 180 185 190Gln Asn Tyr Pro Gln Cys Val Asn Ile Ala Val Thr Gly Ser Gly Thr 195 200 205Lys Ala Leu Pro Ala Gly Thr Pro Ala Thr Gln Leu Tyr Lys Pro Thr 210 215 220Asp Pro Gly Ile Leu Phe Asn Pro Tyr Thr Thr Ile Thr Ser Tyr Thr225 230 235 240Ile Pro Gly Pro Ala Leu Trp Gln Gly 24515232PRTMyceliophthora thermophila 15Met Lys Phe Thr Ser Ser Leu Ala Val Leu Ala Ala Ala Gly Ala Gln1 5 10 15Ala His Tyr Thr Phe Pro Arg Ala Gly Thr Gly Gly Ser Leu Ser Gly 20 25 30Glu Trp Glu Val Val Arg Met Thr Glu Asn His Tyr Ser His Gly Pro 35 40 45Val Thr Asp Val Thr Ser Pro Glu Met Thr Cys Tyr Gln Ser Gly Val 50 55 60Gln Gly Ala Pro Gln Thr Val Gln Val Lys Ala Gly Ser Gln Phe Thr65 70 75 80Phe Ser Val Asp Pro Ser Ile Gly His Pro Gly Pro Leu Gln Phe Tyr 85 90 95Met Ala Lys Val Pro Ser Gly Gln Thr Ala Ala Thr Phe Asp Gly Thr 100 105 110Gly Ala Val Trp Phe Lys Ile Tyr Gln Asp Gly Pro Asn Gly Leu Gly 115 120 125Thr Asp Ser Ile Thr Trp Pro Ser Ala Gly Lys Thr Glu Val Ser Val 130 135 140Thr Ile Pro Ser Cys Ile Asp Asp Gly Glu Tyr Leu Leu Arg Val Glu145 150 155 160His Ile Ala Leu His Ser Ala Ser Ser Val Gly Gly Ala Gln Phe Tyr 165 170 175Ile Ala Cys Ala Gln Leu Ser Val Thr Gly Gly Ser Gly Thr Leu Asn 180 185 190Thr Gly Ser Leu Val Ser Leu Pro Gly Ala Tyr Lys Ala Thr Asp Pro 195 200 205Gly Ile Leu Phe Gln Leu Tyr Trp Pro Ile Pro Thr Glu Tyr Ile Asn 210 215 220Pro Gly Pro Ala Pro Val Ser Cys225 23016235PRTMyceliophthora thermophila 16Met Lys Ala Leu Ser Leu Leu Ala Ala Ala Ser Ala Val Ser Ala His1 5 10 15Thr Ile Phe Val Gln Leu Glu Ala Asp Gly Thr Arg Tyr Pro Val Ser 20 25 30Tyr Gly Ile Arg Asp Pro Ser Tyr Asp Gly Pro Ile Thr Asp Val Thr 35 40 45Ser Asn Asp Val Ala Cys Asn Gly Gly Pro Asn Pro Thr Thr Pro Ser 50 55 60Ser Asp Val Ile Thr Val Thr Ala Gly Thr Thr Val Lys Ala Ile Trp65 70 75 80Arg His Thr Leu Gln Ser Gly Pro Asp Asp Val Met Asp Ala Ser His 85 90 95Lys Gly Pro Thr Leu Ala Tyr Leu Lys Lys Val Gly Asp Ala Thr Lys 100 105 110Asp Ser Gly Val Gly Gly Gly Trp Phe Lys Ile Gln Glu Asp Gly Tyr 115 120 125Asn Asn Gly Gln Trp Gly Thr Ser Thr Val Ile Ser Asn Gly Gly Glu 130 135 140His Tyr Ile Asp Ile Pro Ala Cys Ile Pro Glu Gly Gln Tyr Leu Leu145 150 155 160Arg Ala Glu Met Ile Ala Leu His Ala Ala Gly Ser Pro Gly Gly Ala 165 170 175Gln Leu Tyr Met Glu Cys Ala Gln Ile Asn Ile Val Gly Gly Ser Gly 180 185 190Ser Val Pro Ser Ser Thr Val Ser Phe Pro Gly Ala Tyr Ser Pro Asn 195 200 205Asp Pro Gly Leu Leu Ile Asn Ile Tyr Ser Met Ser Pro Ser Ser Ser 210 215 220Tyr Thr Ile Pro Gly Pro Pro Val Phe Lys Cys225 230 23517323PRTMyceliophthora thermophila 17Met Lys Ser Phe Ala Leu Thr Thr Leu Ala Ala Leu Ala Gly Asn Ala1 5 10 15Ala Ala His Ala Thr Phe Gln Ala Leu Trp Val Asp Gly Val Asp Tyr 20 25 30Gly Ala Gln Cys Ala Arg Leu Pro Ala Ser Asn Ser Pro Val Thr Asp 35 40 45Val Thr Ser Asn Ala Ile Arg Cys Asn Ala Asn Pro Ser Pro Ala Arg 50 55 60Gly Lys Cys Pro Val Lys Ala Gly Ser Thr Val Thr Val Glu Met His65 70 75 80Gln Gln Pro Gly Asp Arg Ser Cys Ser Ser Glu Ala Ile Gly Gly Ala 85 90 95His Tyr Gly Pro Val Met Val Tyr Met Ser Lys Val Ser Asp Ala Ala 100 105 110Ser Ala Asp Gly Ser Ser Gly Trp Phe Lys Val Phe Glu Asp Gly Trp 115 120 125Ala Lys Asn Pro Ser Gly Gly Ser Gly Asp Asp Asp Tyr Trp Gly Thr 130 135 140Lys Asp Leu Asn Ser Cys Cys Gly Lys Met Asn Val Lys Ile Pro Ala145 150 155 160Asp Leu Pro Ser Gly Asp Tyr Leu Leu Arg Ala Glu Ala Leu Ala Leu 165 170 175His Thr Ala Gly Ser Ala Gly Gly Ala Gln Phe Tyr Met Thr Cys Tyr 180 185 190Gln Leu Thr Val Thr Gly Ser Gly Ser Ala Ser Pro Pro Thr Val Ser 195 200 205Phe Pro Gly Ala Tyr Lys Ala Thr Asp Pro Gly Ile Leu Val Asn Ile 210 215 220His Ala Pro Leu Ser Gly Tyr Thr Val Pro Gly Pro Ala Val Tyr Ser225 230 235 240Gly Gly Ser Thr Lys Lys Ala Gly Ser Ala Cys Thr Gly Cys Glu Ser 245 250 255Thr Cys Ala Val Gly Ser Gly Pro Thr Ala Thr Val Ser Gln Ser Pro 260 265 270Gly Ser Thr Ala Thr Ser Ala Pro Gly Gly Gly Gly Gly Cys Thr Val 275 280 285Gln Lys Tyr Gln Gln Cys Gly Gly Glu Gly Tyr Thr Gly Cys Thr Asn 290 295 300Cys Ala Ser Gly Ser Thr Cys Ser Ala Val Ser Pro Pro Tyr Tyr Ser305 310 315 320Gln Cys Val18310PRTMyceliophthora thermophila 18Met Lys Pro Phe Ser Leu Val Ala Leu Ala Thr Ala Val Ser Gly His1 5 10 15Ala Ile Phe Gln Arg Val Ser Val Asn Gly Gln Asp Gln Gly Gln Leu 20 25 30Lys Gly Val Arg Ala Pro Ser Ser Asn Ser Pro Ile Gln Asn Val Asn 35 40 45Asp Ala Asn Met Ala Cys Asn Ala Asn Ile Val Tyr His Asp Ser Thr 50 55 60Ile Ile Lys Val Pro Ala Gly Ala Arg Val Gly Ala Trp Trp Gln His65 70 75 80Val Ile Gly Gly Pro Gln Gly Ala Asn Asp Pro Asp Asn Pro Ile Ala 85 90 95Ala Ser His Lys Gly Pro Ile Gln Val Tyr Leu Ala Lys Val Asp Asn 100 105 110Ala Ala Thr Ala Ser Pro Ser Gly Leu Arg Trp Phe Lys Val Ala Glu 115 120 125Arg Gly Leu Asn Asn Gly Val Trp Ala Val Asp Glu Leu Ile Ala Asn 130 135 140Asn Gly Trp His Tyr Phe Asp Leu Pro Ser Cys Val Ala Pro Gly Gln145 150 155 160Tyr Leu Met Arg Val Glu Leu Leu Ala Leu His Ser Ala Ser Ser Pro

165 170 175Gly Gly Ala Gln Phe Tyr Met Gly Cys Ala Gln Ile Glu Val Thr Gly 180 185 190Ser Gly Thr Asn Ser Gly Ser Asp Phe Val Ser Phe Pro Gly Ala Tyr 195 200 205Ser Ala Asn Asp Pro Gly Ile Leu Leu Ser Ile Tyr Asp Ser Ser Gly 210 215 220Lys Pro Thr Asn Gly Gly Arg Ser Tyr Pro Ile Pro Gly Pro Arg Pro225 230 235 240Ile Ser Cys Ser Gly Ser Gly Asp Gly Gly Asn Asn Gly Gly Gly Gly 245 250 255Asp Asp Asn Asn Asn Asn Asn Gly Gly Gly Asn Asn Gly Gly Gly Gly 260 265 270Gly Gly Ser Val Pro Leu Tyr Gly Gln Cys Gly Gly Ile Gly Tyr Thr 275 280 285Gly Pro Thr Thr Cys Ala Gln Gly Thr Cys Lys Val Ser Asn Glu Tyr 290 295 300Tyr Ser Gln Cys Leu Pro305 31019246PRTMyceliophthora thermophila 19Met Lys Leu Ser Leu Phe Ser Val Leu Ala Thr Ala Leu Thr Val Glu1 5 10 15Gly His Ala Ile Phe Gln Lys Val Ser Val Asn Gly Ala Asp Gln Gly 20 25 30Ser Leu Thr Gly Leu Arg Ala Pro Asn Asn Asn Asn Pro Val Gln Asp 35 40 45Val Asn Ser Gln Asp Met Ile Cys Gly Gln Ser Gly Ser Thr Ser Asn 50 55 60Thr Ile Ile Glu Val Lys Ala Gly Asp Arg Ile Gly Ala Trp Tyr Gln65 70 75 80His Val Ile Gly Gly Ala Gln Phe Pro Asn Asp Pro Asp Asn Pro Ile 85 90 95Ala Lys Ser His Lys Gly Pro Val Met Ala Tyr Leu Ala Lys Val Asp 100 105 110Asn Ala Ala Thr Ala Ser Lys Thr Gly Leu Lys Trp Phe Lys Ile Trp 115 120 125Glu Asp Thr Phe Asn Pro Ser Thr Lys Thr Trp Gly Val Asp Asn Leu 130 135 140Ile Asn Asn Asn Gly Trp Val Tyr Phe Asn Leu Pro Gln Cys Ile Ala145 150 155 160Asp Gly Asn Tyr Leu Leu Arg Val Glu Val Leu Ala Leu His Ser Ala 165 170 175Tyr Ser Gln Gly Gln Ala Gln Phe Tyr Gln Ser Cys Ala Gln Ile Asn 180 185 190Val Ser Gly Gly Gly Ser Phe Thr Pro Pro Ser Thr Val Ser Phe Pro 195 200 205Gly Ala Tyr Ser Ala Ser Asp Pro Gly Ile Leu Ile Asn Ile Tyr Gly 210 215 220Ala Thr Gly Gln Pro Asp Asn Asn Gly Gln Pro Tyr Thr Ala Pro Gly225 230 235 240Pro Ala Pro Ile Ser Cys 24520354PRTThermoascus aurantiacus 20Met Ser Phe Ser Lys Ile Ala Ala Ile Thr Gly Ala Ile Thr Tyr Ala1 5 10 15Ser Leu Ala Ala Ala His Gly Tyr Val Thr Gly Ile Val Ala Asp Gly 20 25 30Thr Tyr Tyr Gly Gly Tyr Ile Val Thr Gln Tyr Pro Tyr Met Ser Thr 35 40 45Pro Pro Asp Val Ile Ala Trp Ser Thr Lys Ala Thr Asp Leu Gly Phe 50 55 60Val Asp Pro Ser Ser Tyr Ala Ser Ser Asp Ile Ile Cys His Lys Gly65 70 75 80Ala Glu Pro Gly Ala Leu Ser Ala Lys Val Ala Ala Gly Gly Thr Val 85 90 95Glu Leu Gln Trp Thr Asp Trp Pro Glu Ser His Lys Gly Pro Val Ile 100 105 110Asp Tyr Leu Ala Ala Cys Asn Gly Asp Cys Ser Thr Val Asp Lys Thr 115 120 125Lys Leu Glu Phe Phe Lys Ile Asp Glu Ser Gly Leu Ile Asp Gly Ser 130 135 140Ser Ala Pro Gly Thr Trp Ala Ser Asp Asn Leu Ile Ala Asn Asn Asn145 150 155 160Ser Trp Thr Val Thr Ile Pro Ser Thr Ile Ala Pro Gly Asn Tyr Val 165 170 175Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Thr Asn Gly 180 185 190Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Glu Val Thr Gly Ser Gly 195 200 205Thr Asp Thr Pro Ala Gly Thr Leu Gly Thr Glu Leu Tyr Lys Ala Thr 210 215 220Asp Pro Gly Ile Leu Val Asn Ile Tyr Gln Thr Leu Thr Ser Tyr Asp225 230 235 240Ile Pro Gly Pro Ala Leu Tyr Thr Gly Gly Ser Ser Gly Ser Ser Gly 245 250 255Ser Ser Asn Thr Ala Lys Ala Thr Thr Ser Thr Ala Ser Ser Ser Ile 260 265 270Val Thr Pro Thr Pro Val Asn Asn Pro Thr Val Thr Gln Thr Ala Val 275 280 285Val Asp Val Thr Gln Thr Val Ser Gln Asn Ala Ala Val Ala Thr Thr 290 295 300Thr Pro Ala Ser Thr Ala Val Ala Thr Ala Val Pro Thr Gly Thr Thr305 310 315 320Phe Ser Phe Asp Ser Met Thr Ser Asp Glu Phe Val Ser Leu Met Arg 325 330 335Ala Thr Val Asn Trp Leu Leu Ser Asn Lys Lys His Ala Arg Asp Leu 340 345 350Ser Tyr21250PRTAspergillus fumigatus 21Met Thr Leu Ser Lys Ile Thr Ser Ile Ala Gly Leu Leu Ala Ser Ala1 5 10 15Ser Leu Val Ala Gly His Gly Phe Val Ser Gly Ile Val Ala Asp Gly 20 25 30Lys Tyr Tyr Gly Gly Tyr Leu Val Asn Gln Tyr Pro Tyr Met Ser Asn 35 40 45Pro Pro Asp Thr Ile Ala Trp Ser Thr Thr Ala Thr Asp Leu Gly Phe 50 55 60Val Asp Gly Thr Gly Tyr Gln Ser Pro Asp Ile Ile Cys His Arg Asp65 70 75 80Ala Lys Asn Gly Lys Leu Thr Ala Thr Val Ala Ala Gly Ser Gln Ile 85 90 95Glu Phe Gln Trp Thr Thr Trp Pro Glu Ser His His Gly Pro Leu Ile 100 105 110Thr Tyr Leu Ala Pro Cys Asn Gly Asp Cys Ala Thr Val Asp Lys Thr 115 120 125Thr Leu Lys Phe Val Lys Ile Ala Ala Gln Gly Leu Ile Asp Gly Ser 130 135 140Asn Pro Pro Gly Val Trp Ala Asp Asp Glu Met Ile Ala Asn Asn Asn145 150 155 160Thr Ala Thr Val Thr Ile Pro Ala Ser Tyr Ala Pro Gly Asn Tyr Val 165 170 175Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Leu Asn Gly 180 185 190Ala Gln Asn Tyr Pro Gln Cys Phe Asn Ile Gln Ile Thr Gly Gly Gly 195 200 205Ser Ala Gln Gly Ser Gly Thr Ala Gly Thr Ser Leu Tyr Lys Asn Thr 210 215 220Asp Pro Gly Ile Lys Phe Asp Ile Tyr Ser Asp Leu Ser Gly Gly Tyr225 230 235 240Pro Ile Pro Gly Pro Ala Leu Phe Asn Ala 245 25022322PRTPenicillium pinophilum 22Met Pro Ser Thr Lys Val Ala Ala Leu Ser Ala Val Leu Ala Leu Ala1 5 10 15Ser Thr Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp Gly 20 25 30Lys Ser Tyr Ser Gly Tyr Leu Val Asn Gln Phe Pro Tyr Glu Ser Asn 35 40 45Pro Pro Ala Val Ile Gly Trp Ala Thr Thr Ala Thr Asp Leu Gly Phe 50 55 60Val Ala Pro Ser Glu Tyr Thr Asn Ala Asp Ile Ile Cys His Lys Asn65 70 75 80Ala Thr Pro Gly Ala Leu Ser Ala Pro Val Ala Ala Gly Gly Thr Val 85 90 95Glu Leu Gln Trp Thr Thr Trp Pro Asp Ser His His Gly Pro Val Ile 100 105 110Ser Tyr Leu Ala Asn Cys Asn Gly Asn Cys Ser Thr Val Asp Lys Thr 115 120 125Lys Leu Asp Phe Val Lys Ile Asp Gln Gly Gly Leu Ile Asp Asp Thr 130 135 140Thr Pro Pro Gly Thr Trp Ala Ser Asp Lys Leu Ile Ala Ala Asn Asn145 150 155 160Ser Trp Thr Val Thr Ile Pro Ser Thr Ile Ala Pro Gly Asn Tyr Val 165 170 175Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Ala Asp Gly 180 185 190Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Glu Ile Thr Gly Ser Gly 195 200 205Thr Ala Ala Pro Ser Gly Thr Ala Gly Glu Lys Leu Tyr Thr Ser Thr 210 215 220Asp Pro Gly Ile Leu Val Asn Ile Tyr Gln Ser Leu Ser Thr Tyr Val225 230 235 240Ile Pro Gly Pro Thr Leu Trp Ser Gly Ala Ala Asn Gly Ala Val Ala 245 250 255Thr Gly Ser Ala Thr Ala Val Ala Thr Thr Ala Thr Ala Ser Ala Thr 260 265 270Ala Thr Pro Thr Thr Leu Val Thr Ser Val Ala Pro Ala Ser Ser Thr 275 280 285Phe Ala Thr Ala Val Val Thr Thr Val Ala Pro Ala Val Thr Asp Val 290 295 300Val Thr Val Thr Asp Val Val Thr Val Thr Thr Val Ile Thr Thr Thr305 310 315 320Val Leu23444PRTThermoascus sp. 23Met Leu Ser Phe Ala Ser Ala Lys Ser Ala Val Leu Thr Thr Leu Leu1 5 10 15Leu Leu Gly Ser Ala Gln Ala His Thr Leu Met Thr Thr Leu Phe Val 20 25 30Asp Gly Val Asn Gln Gly Asp Gly Val Cys Ile Arg Met Asn Asn Asn 35 40 45Gly Ser Thr Ala Asn Thr Tyr Ile Gln Pro Val Thr Ser Lys Asp Ile 50 55 60Ala Cys Gly Ile Gln Gly Glu Ile Gly Ala Ala Arg Val Cys Pro Ala65 70 75 80Lys Ala Ser Ser Thr Leu Thr Phe Gln Phe Arg Glu Gln Pro Ser Asn 85 90 95Pro Asn Ser Ala Pro Leu Asp Pro Ser His Lys Gly Pro Ala Ala Val 100 105 110Tyr Leu Lys Lys Val Asp Ser Ala Ile Ala Ser Asn Asn Ala Ala Gly 115 120 125Asp Gly Trp Phe Lys Ile Trp Glu Ser Val Tyr Asp Glu Ser Thr Gly 130 135 140Lys Trp Gly Thr Thr Lys Met Ile Glu Asn Asn Gly His Ile Ser Val145 150 155 160Lys Val Pro Asp Asp Ile Glu Gly Gly Tyr Tyr Leu Ala Arg Thr Glu 165 170 175Leu Leu Ala Leu His Ala Ala Asn Glu Gly Asp Pro Gln Phe Tyr Val 180 185 190Gly Cys Ala Gln Leu Phe Ile Asp Ser Ala Gly Thr Ala Lys Pro Pro 195 200 205Thr Val Ser Ile Gly Glu Gly Thr Tyr Asp Leu Ser Met Pro Ala Met 210 215 220Thr Tyr Asn Ile Tyr Gln Thr Pro Leu Ala Leu Pro Tyr Pro Met Tyr225 230 235 240Gly Pro Pro Val Tyr Thr Pro Gly Ser Gly Ser Gly Ser Gly Ser Gly 245 250 255Ser Gly Ser Ala Ser Ala Thr Arg Ser Ser Ala Ile Pro Thr Ala Thr 260 265 270Ala Val Thr Asp Cys Ser Ser Glu Glu Asp Arg Glu Asp Ser Val Met 275 280 285Ala Thr Gly Val Pro Val Ala Arg Ser Thr Leu Arg Thr Trp Val Asp 290 295 300Arg Leu Ser Trp His Gly Lys Ala Arg Glu Asn Val Lys Pro Ala Ala305 310 315 320Arg Arg Ser Ala Leu Val Gln Thr Glu Gly Leu Lys Pro Glu Gly Cys 325 330 335Ile Phe Val Asn Gly Asn Trp Cys Gly Phe Glu Val Pro Asp Tyr Asn 340 345 350Asp Ala Glu Ser Cys Trp Ala Ala Ser Asp Asn Cys Trp Lys Gln Ser 355 360 365Asp Ser Cys Trp Asn Gln Thr Gln Pro Thr Gly Tyr Asn Asn Cys Gln 370 375 380Ile Trp Gln Asp Gln Lys Cys Lys Pro Ile Gln Asp Ser Cys Ser Gln385 390 395 400Ser Asn Pro Thr Gly Pro Pro Asn Lys Gly Lys Asp Ile Thr Pro Thr 405 410 415Trp Pro Pro Leu Glu Gly Ser Met Lys Thr Phe Thr Lys Arg Thr Val 420 425 430Ser Tyr Arg Asp Trp Ile Met Lys Arg Lys Gly Ala 435 44024253PRTPenicillium sp. 24Met Leu Ser Ser Thr Thr Arg Thr Leu Ala Phe Thr Gly Leu Ala Gly1 5 10 15Leu Leu Ser Ala Pro Leu Val Lys Ala His Gly Phe Val Gln Gly Ile 20 25 30Val Ile Gly Asp Gln Phe Tyr Ser Gly Tyr Ile Val Asn Ser Phe Pro 35 40 45Tyr Glu Ser Asn Pro Pro Pro Val Ile Gly Trp Ala Thr Thr Ala Thr 50 55 60Asp Leu Gly Phe Val Asp Gly Thr Gly Tyr Gln Gly Pro Asp Ile Ile65 70 75 80Cys His Arg Asn Ala Thr Pro Ala Pro Leu Thr Ala Pro Val Ala Ala 85 90 95Gly Gly Thr Val Glu Leu Gln Trp Thr Pro Trp Pro Asp Ser His His 100 105 110Gly Pro Val Ile Thr Tyr Leu Ala Pro Cys Asn Gly Asn Cys Ser Thr 115 120 125Val Asp Lys Thr Thr Leu Glu Phe Phe Lys Ile Asp Gln Gln Gly Leu 130 135 140Ile Asp Asp Thr Ser Pro Pro Gly Thr Trp Ala Ser Asp Asn Leu Ile145 150 155 160Ala Asn Asn Asn Ser Trp Thr Val Thr Ile Pro Asn Ser Val Ala Pro 165 170 175Gly Asn Tyr Val Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Asn 180 185 190Asn Lys Asp Gly Ala Gln Asn Tyr Pro Gln Cys Ile Asn Ile Glu Val 195 200 205Thr Gly Gly Gly Ser Asp Ala Pro Glu Gly Thr Leu Gly Glu Asp Leu 210 215 220Tyr His Asp Thr Asp Pro Gly Ile Leu Val Asp Ile Tyr Glu Pro Ile225 230 235 240Ala Thr Tyr Thr Ile Pro Gly Pro Pro Glu Pro Thr Phe 245 25025223PRTThielavia terrestris 25Met Lys Leu Ser Ser Gln Leu Ala Ala Leu Thr Leu Ala Ala Ala Ser1 5 10 15Val Ser Gly His Tyr Ile Phe Glu Gln Ile Ala His Gly Gly Thr Lys 20 25 30Phe Pro Pro Tyr Glu Tyr Ile Arg Arg Asn Thr Asn Tyr Asn Ser Pro 35 40 45Val Thr Ser Leu Ser Ser Asn Asp Leu Arg Cys Asn Val Gly Gly Glu 50 55 60Thr Ala Gly Asn Thr Thr Val Leu Asp Val Lys Ala Gly Asp Ser Phe65 70 75 80Thr Phe Tyr Ser Asp Val Ala Val Tyr His Gln Gly Pro Ile Ser Leu 85 90 95Tyr Met Ser Lys Ala Pro Gly Ser Val Val Asp Tyr Asp Gly Ser Gly 100 105 110Asp Trp Phe Lys Ile His Asp Trp Gly Pro Thr Phe Ser Asn Gly Gln 115 120 125Ala Ser Trp Pro Leu Arg Asp Asn Tyr Gln Tyr Asn Ile Pro Thr Cys 130 135 140Ile Pro Asn Gly Glu Tyr Leu Leu Arg Ile Gln Ser Leu Ala Ile His145 150 155 160Asn Pro Gly Ala Thr Pro Gln Phe Tyr Ile Ser Cys Ala Gln Val Arg 165 170 175Val Ser Gly Gly Gly Ser Ala Ser Pro Ser Pro Thr Ala Lys Ile Pro 180 185 190Gly Ala Phe Lys Ala Thr Asp Pro Gly Tyr Thr Ala Asn Ile Tyr Asn 195 200 205Asn Phe His Ser Tyr Thr Val Pro Gly Pro Ala Val Phe Gln Cys 210 215 22026246PRTThielavia terrestris 26Met Lys Phe Ser Leu Val Ser Leu Leu Ala Tyr Gly Leu Ser Val Glu1 5 10 15Ala His Ser Ile Phe Gln Arg Val Ser Val Asn Gly Gln Asp Gln Gly 20 25 30Leu Leu Thr Gly Leu Arg Ala Pro Ser Asn Asn Asn Pro Val Gln Asp 35 40 45Val Asn Ser Gln Asn Met Ile Cys Gly Gln Ser Gly Ser Lys Ser Gln 50 55 60Thr Val Ile Asn Val Lys Ala Gly Asp Arg Ile Gly Ser Leu Trp Gln65 70 75 80His Val Ile Gly Gly Ala Gln Phe Ser Gly Asp Pro Asp Asn Pro Ile 85 90 95Ala His Ser His Lys Gly Pro Val Met Ala Tyr Leu Ala Lys Val Asp 100 105 110Asn Ala Ala Ser Ala Ser Gln Thr Gly Leu Lys Trp Phe Lys Ile Trp 115 120 125Gln Asp Gly Phe Asp Thr Ser Ser Lys Thr Trp Gly Val Asp Asn Leu 130 135 140Ile Lys Asn Asn Gly Trp Val Tyr Phe His Leu Pro Gln Cys Leu Ala145 150 155 160Pro Gly Gln Tyr Leu Leu Arg Val Glu Val Leu Ala Leu His Ser Ala 165 170 175Tyr Gln Gln Gly Gln Ala Gln Phe Tyr Gln Ser Cys Ala Gln Ile Asn 180 185 190Val Ser Gly Ser Gly Ser Phe Ser Pro Ser Gln Thr Val Ser Ile Pro 195 200 205Gly Val Tyr Ser

Ala Thr Asp Pro Ser Ile Leu Ile Asn Ile Tyr Gly 210 215 220Ser Thr Gly Gln Pro Asp Asn Gly Gly Lys Ala Tyr Asn Pro Pro Gly225 230 235 240Pro Ala Pro Ile Ser Cys 24527334PRTThielavia terrestris 27Met Arg Thr Thr Phe Ala Ala Ala Leu Ala Ala Phe Ala Ala Gln Glu1 5 10 15Val Ala Gly His Ala Ile Phe Gln Gln Leu Trp His Gly Ser Ser Cys 20 25 30Val Arg Met Pro Leu Ser Asn Ser Pro Val Thr Asn Val Gly Ser Arg 35 40 45Asp Met Ile Cys Asn Ala Gly Thr Arg Pro Val Ser Gly Lys Cys Pro 50 55 60Val Lys Ala Gly Gly Thr Val Thr Val Glu Met His Gln Gln Pro Gly65 70 75 80Asp Arg Ser Cys Asn Asn Glu Ala Ile Gly Gly Ala His Trp Gly Pro 85 90 95Val Gln Val Tyr Leu Ser Lys Val Glu Asp Ala Ser Thr Ala Asp Gly 100 105 110Ser Thr Gly Trp Phe Lys Ile Phe Ala Asp Thr Trp Ser Lys Lys Ala 115 120 125Gly Ser Ser Val Gly Asp Asp Asp Asn Trp Gly Thr Arg Asp Leu Asn 130 135 140Ala Cys Cys Gly Lys Met Gln Val Lys Ile Pro Ala Asp Ile Pro Ser145 150 155 160Gly Asp Tyr Leu Leu Arg Ala Glu Ala Leu Ala Leu His Thr Ala Gly 165 170 175Gln Val Gly Gly Ala Gln Phe Tyr Met Ser Cys Tyr Gln Ile Thr Val 180 185 190Ser Gly Gly Gly Ser Ala Ser Pro Ala Thr Val Lys Phe Pro Gly Ala 195 200 205Tyr Ser Ala Asn Asp Pro Gly Ile His Ile Asn Ile His Ala Ala Val 210 215 220Ser Asn Tyr Val Ala Pro Gly Pro Ala Val Tyr Ser Gly Gly Thr Thr225 230 235 240Lys Val Ala Gly Ser Gly Cys Gln Gly Cys Glu Asn Thr Cys Lys Val 245 250 255Gly Ser Ser Pro Thr Ala Thr Ala Pro Ser Gly Lys Ser Gly Ala Gly 260 265 270Ser Asp Gly Gly Ala Gly Thr Asp Gly Gly Ser Ser Ser Ser Ser Pro 275 280 285Asp Thr Gly Ser Ala Cys Ser Val Gln Ala Tyr Gly Gln Cys Gly Gly 290 295 300Asn Gly Tyr Ser Gly Cys Thr Gln Cys Ala Pro Gly Tyr Thr Cys Lys305 310 315 320Ala Val Ser Pro Pro Tyr Tyr Ser Gln Cys Ala Pro Ser Ser 325 33028227PRTThielavia terrestris 28Met Lys Leu Ser Val Ala Ile Ala Val Leu Ala Ser Ala Leu Ala Glu1 5 10 15Ala His Tyr Thr Phe Pro Ser Ile Gly Asn Thr Ala Asp Trp Gln Tyr 20 25 30Val Arg Ile Thr Thr Asn Tyr Gln Ser Asn Gly Pro Val Thr Asp Val 35 40 45Thr Ser Asp Gln Ile Arg Cys Tyr Glu Arg Asn Pro Gly Thr Gly Ala 50 55 60Gln Gly Ile Tyr Asn Val Thr Ala Gly Gln Thr Ile Asn Tyr Asn Ala65 70 75 80Lys Ala Ser Ile Ser His Pro Gly Pro Met Ser Phe Tyr Ile Ala Lys 85 90 95Val Pro Ala Gly Gln Thr Ala Ala Thr Trp Asp Gly Lys Gly Ala Val 100 105 110Trp Thr Lys Ile Tyr Gln Asp Met Pro Lys Phe Gly Ser Ser Leu Thr 115 120 125Trp Pro Thr Met Gly Ala Lys Ser Val Pro Val Thr Ile Pro Arg Cys 130 135 140Leu Gln Asn Gly Asp Tyr Leu Leu Arg Ala Glu His Ile Ala Leu His145 150 155 160Ser Ala Ser Ser Val Gly Gly Ala Gln Phe Tyr Leu Ser Cys Ala Gln 165 170 175Leu Thr Val Ser Gly Gly Ser Gly Thr Trp Asn Pro Lys Asn Arg Val 180 185 190Ser Phe Pro Gly Ala Tyr Lys Ala Thr Asp Pro Gly Ile Leu Ile Asn 195 200 205Ile Tyr Tyr Pro Val Pro Thr Ser Tyr Ser Pro Pro Gly Pro Pro Ala 210 215 220Glu Thr Cys22529368PRTThielavia terrestris 29Met Pro Ser Phe Ala Ser Lys Thr Leu Leu Ser Thr Leu Ala Gly Ala1 5 10 15Ala Ser Val Ala Ala His Gly His Val Ser Asn Ile Val Ile Asn Gly 20 25 30Val Ser Tyr Gln Gly Tyr Asp Pro Thr Ser Phe Pro Tyr Met Gln Asn 35 40 45Pro Pro Ile Val Val Gly Trp Thr Ala Ala Asp Thr Asp Asn Gly Phe 50 55 60Val Ala Pro Asp Ala Phe Ala Ser Gly Asp Ile Ile Cys His Lys Asn65 70 75 80Ala Thr Asn Ala Lys Gly His Ala Val Val Ala Ala Gly Asp Lys Ile 85 90 95Phe Ile Gln Trp Asn Thr Trp Pro Glu Ser His His Gly Pro Val Ile 100 105 110Asp Tyr Leu Ala Ser Cys Gly Ser Ala Ser Cys Glu Thr Val Asp Lys 115 120 125Thr Lys Leu Glu Phe Phe Lys Ile Asp Glu Val Gly Leu Val Asp Gly 130 135 140Ser Ser Ala Pro Gly Val Trp Gly Ser Asp Gln Leu Ile Ala Asn Asn145 150 155 160Asn Ser Trp Leu Val Glu Ile Pro Pro Thr Ile Ala Pro Gly Asn Tyr 165 170 175Val Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Glu Asn Ala Asp 180 185 190Gly Ala Gln Asn Tyr Pro Gln Cys Phe Asn Leu Gln Ile Thr Gly Thr 195 200 205Gly Thr Ala Thr Pro Ser Gly Val Pro Gly Thr Ser Leu Tyr Thr Pro 210 215 220Thr Asp Pro Gly Ile Leu Val Asn Ile Tyr Ser Ala Pro Ile Thr Tyr225 230 235 240Thr Val Pro Gly Pro Ala Leu Ile Ser Gly Ala Val Ser Ile Ala Gln 245 250 255Ser Ser Ser Ala Ile Thr Ala Ser Gly Thr Ala Leu Thr Gly Ser Ala 260 265 270Thr Ala Pro Ala Ala Ala Ala Ala Thr Thr Thr Ser Thr Thr Asn Ala 275 280 285Ala Ala Ala Ala Thr Ser Ala Ala Ala Ala Ala Gly Thr Ser Thr Thr 290 295 300Thr Thr Ser Ala Ala Ala Val Val Gln Thr Ser Ser Ser Ser Ser Ser305 310 315 320Ala Pro Ser Ser Ala Ala Ala Ala Ala Thr Thr Thr Ala Ala Ala Ser 325 330 335Ala Arg Pro Thr Gly Cys Ser Ser Gly Arg Ser Arg Lys Gln Pro Arg 340 345 350Arg His Ala Arg Asp Met Val Val Ala Arg Gly Ala Glu Glu Ala Asn 355 360 36530330PRTThielavia terrestris 30Met Pro Pro Ala Leu Pro Gln Leu Leu Thr Thr Val Leu Thr Ala Leu1 5 10 15Thr Leu Gly Ser Thr Ala Leu Ala His Ser His Leu Ala Tyr Ile Ile 20 25 30Val Asn Gly Lys Leu Tyr Gln Gly Phe Asp Pro Arg Pro His Gln Ala 35 40 45Asn Tyr Pro Ser Arg Val Gly Trp Ser Thr Gly Ala Val Asp Asp Gly 50 55 60Phe Val Thr Pro Ala Asn Tyr Ser Thr Pro Asp Ile Ile Cys His Ile65 70 75 80Ala Gly Thr Ser Pro Ala Gly His Ala Pro Val Arg Pro Gly Asp Arg 85 90 95Ile His Val Gln Trp Asn Gly Trp Pro Val Gly His Ile Gly Pro Val 100 105 110Leu Ser Tyr Leu Ala Arg Cys Glu Ser Asp Thr Gly Cys Thr Gly Gln 115 120 125Asn Lys Thr Ala Leu Arg Trp Thr Lys Ile Asp Asp Ser Ser Pro Thr 130 135 140Met Gln Asn Val Ala Gly Ala Gly Thr Gln Gly Glu Gly Thr Pro Gly145 150 155 160Lys Arg Trp Ala Thr Asp Val Leu Ile Ala Ala Asn Asn Ser Trp Gln 165 170 175Val Ala Val Pro Ala Gly Leu Pro Thr Gly Ala Tyr Val Leu Arg Asn 180 185 190Glu Ile Ile Ala Leu His Tyr Ala Ala Arg Lys Asn Gly Ala Gln Asn 195 200 205Tyr Pro Leu Cys Met Asn Leu Trp Val Asp Ala Ser Gly Asp Asn Ser 210 215 220Ser Val Ala Ala Thr Thr Ala Ala Val Thr Ala Gly Gly Leu Gln Met225 230 235 240Asp Ala Tyr Asp Ala Arg Gly Phe Tyr Lys Glu Asn Asp Pro Gly Val 245 250 255Leu Val Asn Val Thr Ala Ala Leu Ser Ser Tyr Val Val Pro Gly Pro 260 265 270Thr Val Ala Ala Gly Ala Thr Pro Val Pro Tyr Ala Gln Gln Ser Pro 275 280 285Ser Val Ser Thr Ala Ala Gly Thr Pro Val Val Val Thr Arg Thr Ser 290 295 300Glu Thr Ala Pro Tyr Thr Gly Ala Met Thr Pro Thr Val Ala Ala Arg305 310 315 320Met Lys Gly Arg Gly Tyr Asp Arg Arg Gly 325 33031236PRTThielavia terrestris 31Met Lys Thr Phe Thr Ala Leu Leu Ala Ala Ala Gly Leu Val Ala Gly1 5 10 15His Gly Tyr Val Asp Asn Ala Thr Ile Gly Gly Gln Phe Tyr Gln Asn 20 25 30Pro Ala Val Leu Thr Phe Phe Gln Pro Asp Arg Val Ser Arg Ser Ile 35 40 45Pro Gly Asn Gly Pro Val Thr Asp Val Thr Leu Ile Asp Leu Gln Cys 50 55 60Asn Ala Asn Ser Thr Pro Ala Lys Leu His Ala Thr Ala Ala Ala Gly65 70 75 80Ser Asp Val Ile Leu Arg Trp Thr Leu Trp Pro Glu Ser His Val Gly 85 90 95Pro Val Ile Thr Tyr Met Ala Arg Cys Pro Asp Thr Gly Cys Gln Asp 100 105 110Trp Met Pro Gly Thr Ser Ala Val Trp Phe Lys Ile Lys Glu Gly Gly 115 120 125Arg Asp Gly Thr Ser Asn Thr Trp Ala Asp Thr Pro Leu Met Thr Ala 130 135 140Pro Thr Ser Tyr Thr Tyr Thr Ile Pro Ser Cys Leu Lys Lys Gly Tyr145 150 155 160Tyr Leu Val Arg His Glu Ile Ile Ala Leu His Ala Ala Tyr Thr Tyr 165 170 175Pro Gly Ala Gln Phe Tyr Pro Gly Cys His Gln Leu Asn Val Thr Gly 180 185 190Gly Gly Ser Thr Val Pro Ser Ser Gly Leu Val Ala Phe Pro Gly Ala 195 200 205Tyr Lys Gly Ser Asp Pro Gly Ile Thr Tyr Asp Ala Tyr Lys Ala Gln 210 215 220Thr Tyr Gln Ile Pro Gly Pro Ala Val Phe Thr Cys225 230 23532250PRTThielavia terrestris 32Met Ala Leu Leu Leu Leu Ala Gly Leu Ala Ile Leu Ala Gly Pro Ala1 5 10 15His Ala His Gly Gly Leu Ala Asn Tyr Thr Val Gly Asn Thr Trp Tyr 20 25 30Arg Gly Tyr Asp Pro Phe Thr Pro Ala Ala Asp Gln Ile Gly Gln Pro 35 40 45Trp Met Ile Gln Arg Ala Trp Asp Ser Ile Asp Pro Ile Phe Ser Val 50 55 60Asn Asp Lys Ala Leu Ala Cys Asn Thr Pro Ala Thr Ala Pro Thr Ser65 70 75 80Tyr Ile Pro Ile Arg Ala Gly Glu Asn Ile Thr Ala Val Tyr Trp Tyr 85 90 95Trp Leu His Pro Val Gly Pro Met Thr Ala Trp Leu Ala Arg Cys Asp 100 105 110Gly Asp Cys Arg Asp Ala Asp Val Asn Glu Ala Arg Trp Phe Lys Ile 115 120 125Trp Glu Ala Gly Leu Leu Ser Gly Pro Asn Leu Ala Glu Gly Met Trp 130 135 140Tyr Gln Lys Ala Phe Gln Asn Trp Asp Gly Ser Pro Asp Leu Trp Pro145 150 155 160Val Thr Ile Pro Ala Gly Leu Lys Ser Gly Leu Tyr Met Ile Arg His 165 170 175Glu Ile Leu Ser Ile His Val Glu Asp Lys Pro Gln Phe Tyr Pro Glu 180 185 190Cys Ala His Leu Asn Val Thr Gly Gly Gly Asp Leu Leu Pro Pro Asp 195 200 205Glu Phe Leu Val Lys Phe Pro Gly Ala Tyr Lys Glu Asp Asn Pro Ser 210 215 220Ile Lys Ile Asn Ile Tyr Ser Asp Gln Tyr Ala Asn Thr Thr Asn Tyr225 230 235 240Thr Ile Pro Gly Gly Pro Ile Trp Asp Gly 245 25033478PRTThielavia terrestris 33Met Met Pro Ser Leu Val Arg Phe Ser Met Gly Leu Ala Thr Ala Phe1 5 10 15Ala Ser Leu Ser Thr Ala His Thr Val Phe Thr Thr Leu Phe Ile Asn 20 25 30Gly Val Asp Gln Gly Asp Gly Thr Cys Ile Arg Met Ala Lys Lys Gly 35 40 45Ser Val Cys Thr His Pro Ile Ala Gly Gly Leu Asp Ser Pro Asp Met 50 55 60Ala Cys Gly Arg Asp Gly Gln Gln Ala Val Ala Phe Thr Cys Pro Ala65 70 75 80Pro Ala Gly Ser Lys Leu Ser Phe Glu Phe Arg Met Trp Ala Asp Ala 85 90 95Ser Gln Pro Gly Ser Ile Asp Pro Ser His Leu Gly Ser Thr Ala Ile 100 105 110Tyr Leu Lys Gln Val Ser Asn Ile Ser Ser Asp Ser Ala Ala Gly Pro 115 120 125Gly Trp Phe Lys Ile Tyr Ala Glu Gly Tyr Asp Thr Ala Ala Lys Lys 130 135 140Trp Ala Thr Glu Lys Leu Ile Asp Asn Gly Gly Leu Leu Ser Ile Glu145 150 155 160Leu Pro Pro Thr Leu Pro Ala Gly Tyr Tyr Leu Ala Arg Ser Glu Ile 165 170 175Val Thr Ile Gln Asn Val Thr Asn Asp His Val Asp Pro Gln Phe Tyr 180 185 190Val Gly Cys Ala Gln Leu Phe Val Gln Gly Pro Pro Thr Thr Pro Thr 195 200 205Val Pro Pro Asp Arg Leu Val Ser Ile Pro Gly His Val His Ala Ser 210 215 220Asp Pro Gly Leu Thr Phe Asn Ile Trp Arg Asp Asp Pro Ser Lys Thr225 230 235 240Ala Tyr Thr Val Val Gly Pro Ala Pro Phe Ser Pro Thr Ala Ala Pro 245 250 255Thr Pro Thr Ser Thr Asn Thr Asn Gly Gln Gln Gln Gln Gln Gln Gln 260 265 270Gln Ala Ile Lys Gln Thr Asp Gly Val Ile Pro Ala Asp Cys Gln Leu 275 280 285Lys Asn Ala Asn Trp Cys Gly Ala Glu Val Pro Ala Tyr Ala Asp Glu 290 295 300Ala Gly Cys Trp Ala Ser Ser Ala Asp Cys Phe Ala Gln Leu Asp Ala305 310 315 320Cys Tyr Thr Ser Ala Pro Pro Thr Gly Ser Arg Gly Cys Arg Leu Trp 325 330 335Glu Asp Trp Cys Thr Gly Ile Gln Gln Gly Cys Arg Ala Gly Arg Trp 340 345 350Arg Gly Pro Pro Pro Phe His Gly Glu Gly Ala Ala Ala Glu Thr Ala 355 360 365Ser Ala Gly Arg Gly Gly Ala Arg Ile Ala Ala Val Ala Gly Cys Gly 370 375 380Gly Gly Thr Gly Asp Met Val Glu Glu Val Phe Leu Phe Tyr Trp Asp385 390 395 400Ala Cys Ser Gly Trp Arg Arg Ser Arg Gly Gly Gly Ser Ile Leu Ala 405 410 415Arg Leu Ile Leu His Val Leu Leu Pro Leu Leu Arg Pro Arg Arg Ala 420 425 430Pro Arg Val His Leu Leu Leu Phe His Leu Tyr Leu Asn Phe Cys Tyr 435 440 445Pro Gly Thr Ser Gly Phe Tyr Asn Arg Leu Ser Ile Lys Leu Gly Ile 450 455 460Trp Pro Ser Lys Met Ser Pro Asp Val Ala His Tyr Val Lys465 470 47534230PRTThielavia terrestris 34Met Gln Leu Leu Val Gly Leu Leu Leu Ala Ala Val Ala Ala Arg Ala1 5 10 15His Tyr Thr Phe Pro Arg Leu Val Val Asn Gly Gln Pro Glu Asp Lys 20 25 30Asp Trp Ser Val Thr Arg Met Thr Lys Asn Ala Gln Ser Lys Gln Gly 35 40 45Val Gln Asp Pro Thr Ser Pro Asp Ile Arg Cys Tyr Thr Ser Gln Thr 50 55 60Ala Pro Asn Val Ala Thr Val Pro Ala Gly Ala Thr Val His Tyr Ile65 70 75 80Ser Thr Gln Gln Ile Asn His Pro Gly Pro Thr Gln Tyr Tyr Leu Ala 85 90 95Lys Val Pro Ala Gly Ser Ser Ala Lys Thr Trp Asp Gly Ser Gly Ala 100 105 110Val Trp Phe Lys Ile Ser Thr Thr Met Pro Tyr Leu Asp Asn Asn Lys 115 120 125Gln Leu Val Trp Pro Asn Gln Asn Thr Tyr Thr Thr Val Asn Thr Thr 130 135 140Ile Pro Ala Asp Thr Pro Ser Gly Glu Tyr Leu Leu Arg Val Glu Gln145 150 155 160Ile Ala Leu His Leu Ala Ser Gln Pro Asn Gly Ala Gln Phe Tyr Leu 165 170 175Ala Cys Ser Gln Ile Gln Ile Thr Gly Gly Gly Asn Gly Thr Pro Gly

180 185 190Pro Leu Val Ala Leu Pro Gly Ala Tyr Lys Ser Asn Asp Pro Gly Ile 195 200 205Leu Val Asn Ile Tyr Ser Met Gln Pro Gly Asp Tyr Lys Pro Pro Gly 210 215 220Pro Pro Val Trp Ser Gly225 23035257PRTThielavia terrestris 35Met Lys Leu Tyr Leu Ala Ala Phe Leu Gly Ala Val Ala Thr Pro Gly1 5 10 15Ala Phe Ala His Gln Ile His Gly Ile Leu Leu Val Asn Gly Thr Glu 20 25 30Thr Pro Glu Trp Lys Tyr Val Arg Asp Val Ala Trp Glu Gly Ala Tyr 35 40 45Glu Pro Glu Lys Tyr Pro Asn Thr Glu Phe Phe Lys Thr Pro Pro Gln 50 55 60Thr Asp Ile Asn Asn Pro Asn Ile Thr Cys Gly Arg Asn Ala Phe Asp65 70 75 80Ser Ala Ser Lys Thr Glu Thr Ala Asp Ile Leu Ala Gly Ser Glu Val 85 90 95Gly Phe Arg Val Ser Trp Asp Gly Asn Gly Lys Tyr Gly Val Phe Trp 100 105 110His Pro Gly Pro Gly Gln Ile Tyr Leu Ser Arg Ala Pro Asn Asp Asp 115 120 125Leu Glu Asp Tyr Arg Gly Asp Gly Asp Trp Phe Lys Ile Ala Thr Gly 130 135 140Ala Ala Val Ser Asn Thr Glu Trp Leu Leu Trp Asn Lys His Asp Phe145 150 155 160Asn Phe Thr Ile Pro Lys Thr Thr Pro Pro Gly Lys Tyr Leu Met Arg 165 170 175Ile Glu Gln Phe Met Pro Ser Thr Val Glu Tyr Ser Gln Trp Tyr Val 180 185 190Asn Cys Ala His Val Asn Ile Ile Gly Pro Gly Gly Gly Thr Pro Thr 195 200 205Gly Phe Ala Arg Phe Pro Gly Thr Tyr Thr Val Asp Asp Pro Gly Ile 210 215 220Lys Val Pro Leu Asn Gln Ile Val Asn Ser Gly Glu Leu Pro Gln Asp225 230 235 240Gln Leu Arg Leu Leu Glu Tyr Lys Pro Pro Gly Pro Ala Leu Trp Thr 245 250 255Gly36251PRTThermoascus crustaceus 36Met Ala Phe Ser Gln Ile Met Ala Ile Thr Gly Val Phe Leu Ala Ser1 5 10 15Ala Ser Leu Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp 20 25 30Gly Lys Ser Tyr Gly Gly Tyr Ile Val Asn Gln Tyr Pro Tyr Met Ser 35 40 45Asp Pro Pro Glu Val Val Gly Trp Ser Thr Thr Ala Thr Asp Leu Gly 50 55 60Phe Val Asp Gly Thr Gly Tyr Gln Gly Pro Asp Ile Ile Cys His Arg65 70 75 80Gly Ala Lys Pro Ala Ala Leu Thr Ala Gln Val Ala Ala Gly Gly Thr 85 90 95Val Lys Leu Glu Trp Thr Pro Trp Pro Asp Ser His His Gly Pro Val 100 105 110Ile Asn Tyr Leu Ala Pro Cys Asn Gly Asp Cys Ser Thr Val Asp Lys 115 120 125Thr Gln Leu Lys Phe Phe Lys Ile Ala Gln Ala Gly Leu Ile Asp Asp 130 135 140Asn Ser Pro Pro Gly Ile Trp Ala Ser Asp Asn Leu Ile Ala Ala Asn145 150 155 160Asn Ser Trp Thr Val Thr Ile Pro Thr Thr Thr Ala Pro Gly Asn Tyr 165 170 175Val Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Lys Asp 180 185 190Gly Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Lys Val Thr Gly Asn 195 200 205Gly Ser Gly Asn Pro Pro Ala Gly Ala Leu Gly Thr Ala Leu Tyr Lys 210 215 220Asp Thr Asp Pro Gly Ile Leu Ile Asn Ile Tyr Gln Lys Leu Ser Ser225 230 235 240Tyr Val Ile Pro Gly Pro Ala Leu Tyr Thr Gly 245 25037349PRTThermoascus crustaceus 37Met Ser Phe Ser Lys Ile Leu Ala Ile Ala Gly Ala Ile Thr Tyr Ala1 5 10 15Ser Ser Ala Ala Ala His Gly Tyr Val Gln Gly Ile Val Val Asp Gly 20 25 30Ser Tyr Tyr Gly Gly Tyr Met Val Thr Gln Tyr Pro Tyr Thr Ala Gln 35 40 45Pro Pro Glu Leu Ile Ala Trp Ser Thr Lys Ala Thr Asp Leu Gly Phe 50 55 60Val Asp Gly Ser Gly Tyr Thr Ser Pro Asp Ile Ile Cys His Lys Gly65 70 75 80Ala Glu Pro Gly Ala Gln Ser Ala Lys Val Ala Ala Gly Gly Thr Val 85 90 95Glu Leu Gln Trp Thr Ala Trp Pro Glu Ser His Lys Gly Pro Val Ile 100 105 110Asp Tyr Leu Ala Ala Cys Asp Gly Asp Cys Ser Ser Val Asp Lys Thr 115 120 125Ala Leu Lys Phe Phe Lys Ile Asp Glu Ser Gly Leu Ile Asp Gly Asn 130 135 140Gly Ala Gly Thr Trp Ala Ser Asp Thr Leu Ile Lys Asn Asn Asn Ser145 150 155 160Trp Thr Val Thr Ile Pro Ser Thr Ile Ala Ser Gly Asn Tyr Val Leu 165 170 175Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Lys Asp Gly Ala 180 185 190Gln Asn Tyr Pro Gln Cys Ile Asn Leu Glu Val Thr Gly Ser Gly Thr 195 200 205Glu Asn Pro Ala Gly Thr Leu Gly Thr Ala Leu Tyr Thr Asp Thr Asp 210 215 220Pro Gly Leu Leu Val Asn Ile Tyr Gln Gly Leu Ser Asn Tyr Ser Ile225 230 235 240Pro Gly Pro Ala Leu Tyr Ser Gly Asn Ser Asp Asn Ala Gly Ser Leu 245 250 255Asn Pro Thr Thr Thr Pro Ser Ile Gln Asn Ala Ala Ala Ala Pro Ser 260 265 270Thr Ser Thr Ala Ser Val Val Thr Asp Ser Ser Ser Ala Thr Gln Thr 275 280 285Ala Ser Val Ala Ala Thr Thr Pro Ala Ser Thr Ser Ala Val Thr Ala 290 295 300Ser Pro Ala Pro Asp Thr Gly Ser Asp Val Thr Lys Tyr Leu Asp Ser305 310 315 320Met Ser Ser Asp Glu Val Leu Thr Leu Val Arg Gly Thr Leu Ser Trp 325 330 335Leu Val Ser Asn Lys Lys His Ala Arg Asp Leu Ser His 340 34538436PRTThermoascus crustaceus 38Met Leu Ser Phe Ile Pro Thr Lys Ser Ala Ala Leu Thr Thr Leu Leu1 5 10 15Leu Leu Gly Thr Ala His Ala His Thr Leu Met Thr Thr Met Phe Val 20 25 30Asp Gly Val Asn Gln Gly Asp Gly Val Cys Ile Arg Met Asn Asn Asp 35 40 45Gly Gly Thr Ala Asn Thr Tyr Ile Gln Pro Ile Thr Ser Lys Asp Ile 50 55 60Ala Cys Gly Ile Gln Gly Glu Ile Gly Ala Ser Arg Val Cys Pro Val65 70 75 80Lys Ala Ser Ser Thr Leu Thr Phe Gln Phe Arg Glu Gln Pro Asn Asn 85 90 95Pro Asn Ser Ser Pro Leu Asp Pro Ser His Lys Gly Pro Ala Ala Val 100 105 110Tyr Leu Lys Lys Val Asp Ser Ala Ile Ala Ser Asn Asn Ala Ala Gly 115 120 125Asp Ser Trp Phe Lys Ile Trp Glu Ser Val Tyr Asp Glu Ser Thr Gly 130 135 140Lys Trp Gly Thr Thr Lys Met Ile Glu Asn Asn Gly His Ile Ser Val145 150 155 160Lys Val Pro Asp Asp Ile Glu Gly Gly Tyr Tyr Leu Ala Arg Thr Glu 165 170 175Leu Leu Ala Leu His Ser Ala Asp Gln Gly Asp Pro Gln Phe Tyr Val 180 185 190Gly Cys Ala Gln Leu Phe Ile Asp Ser Asp Gly Thr Ala Lys Pro Pro 195 200 205Thr Val Ser Ile Gly Glu Gly Thr Tyr Asp Leu Ser Met Pro Ala Met 210 215 220Thr Tyr Asn Ile Trp Glu Thr Pro Leu Ala Leu Pro Tyr Pro Met Tyr225 230 235 240Gly Pro Pro Val Tyr Thr Pro Gly Ser Gly Ser Gly Ser Val Arg Ala 245 250 255Thr Ser Ser Ser Ala Val Pro Thr Ala Thr Glu Ser Ser Phe Val Glu 260 265 270Glu Arg Ala Asn Pro Val Thr Ala Asn Ser Val Tyr Ser Ala Arg Gly 275 280 285Lys Phe Lys Thr Trp Ile Asp Lys Leu Ser Trp Arg Gly Lys Val Arg 290 295 300Glu Asn Val Arg Gln Ala Ala Gly Arg Arg Ser Thr Leu Val Gln Thr305 310 315 320Val Gly Leu Lys Pro Lys Gly Cys Ile Phe Val Asn Gly Asn Trp Cys 325 330 335Gly Phe Glu Val Pro Asp Tyr Asn Asp Ala Glu Ser Cys Trp Ala Ala 340 345 350Ser Asp Asn Cys Trp Lys Gln Ser Asp Ala Cys Trp Asn Lys Thr Gln 355 360 365Pro Thr Gly Tyr Asn Asn Cys Gln Ile Trp Gln Asp Lys Lys Cys Lys 370 375 380Val Ile Gln Asp Ser Cys Ser Gly Pro Asn Pro His Gly Pro Pro Asn385 390 395 400Lys Gly Lys Asp Leu Thr Pro Glu Trp Pro Pro Leu Lys Gly Ser Met 405 410 415Asp Thr Phe Ser Lys Arg Thr Ile Gly Tyr Arg Asp Trp Ile Val Arg 420 425 430Arg Arg Gly Ala 435

Claims

1. A method for treating dissolving pulp, comprising a step of subjecting the dissolving pulp to a lytic polysaccharide monooxygenase.

2. The method according to claim 1, further comprising a step of subjecting the dissolving pulp to a cellulase, pre:erably an endoglucanase.

3. The method according to claim 2, wherein the step of subjecting the dissolving pulp to a lytic polysaccharide monooxygenase and the step of subjecting the dissolving pulp to a cellulase are carried out simultaneously.

4. The method according to claim 1, further comprising a step of bleaching the dissolving pulp.

5. The method according to claim 1, further comprising a step of subjecting the dissolving pulp to a lytic polysaccharide monooxygenase and an electron donor thereof, preferably ascorbic acid, gallic acid, pyrogallol or cysteine.

6. The method according to claim 1, wherein the dissolving pulp is an alkaline extracted dissolving pulp; kraft pulp or sulfite pulp.

7. The method according to claim 1, wherein the lytic polysaccharide monooxygenase has a sequence identity of at least 60% to the mature polypeptide of SEQ ID NO: 1 and/or to the mature polypeptide of SEQ ID NO: 2 and/or to the mature polypeptide of SEQ ID NO: 3.

8. The method according to claim 2, wherein the cellulase has a sequence identity of at least 60% to SEQ ID NO: 4, and/or to the mature polypeptide of SEQ ID NO: 5, and/or to the mature polypeptide of SEQ ID NO: 6, and/or to the mature polypeptide of SEQ ID NO: 7.

9. The method according to claim 1, wherein the lytic polysaccharide monooxygenase comprises SEQ ID NO: 1 or the mature polypeptide thereof, SEQ ID NO: 2 or the mature polypeptide thereof, or SEQ ID NO: 3 or the mature polypeptide thereof.

10. The method according to claim 1, wherein concentration of the lytic polysaccharide monooxygenase is from 0.05 mg/kg oven dry pulp to 100000 mg/kg oven dry pulp, optionally a concentration selected from the group consisting of from 0.05 mg/kg oven dry pulp to 250 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 1000 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 2000 mg/kg oven dry pulp, from 1.0 mg/kg oven dry pulp to 5000 mg/kg oven dry pulp, from 5.0 mg/kg oven dry pulp to 10000 mg/kg oven dry pulp, from 10.0 mg/kg oven dry pulp to 15000 mg/kg oven dry pulp, from 15.0 mg/kg oven dry pulp to 20000 mg/kg oven dry pulp, from 20.0 mg/kg oven dry pulp to 30000 mg/kg oven dry pulp, from 30.0 mg/kg oven dry pulp to 40000 mg/kg oven dry pulp, from 40.0 mg/kg oven dry pulp to 60000 mg/kg oven dry pulp, from 60.0 mg/kg oven dry pulp to 80000 mg/kg oven dry pulp, and from 80.0 mg/kg oven dry pulp to 100000 mg/kg oven dry pulp, or any combination of these intervals.

11. The method according to claim 2, wherein concentration of the cellulase is from 0.05 mg/kg oven dry pulp to 100 mg/kg oven dry pulp, optionally a concentration selected from the group consisting of from 0.05 mg/kg oven dry pulp to 80.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 60.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 40.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 20.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 10.0 mg/kg oven dry pulp, from 0.25 mg/kg oven dry pulp to 5.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 85.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 65.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 45.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 25.0 mg/kg oven dry pulp, from 0.50 mg/kg oven dry pulp to 15.0 mg/kg oven dry pulp and from 0.50 mg/kg oven dry pulp to 5.0 mg/kg oven dry pulp or any combination of these intervals.

12. The method according to claim 1, wherein the method results in reduced viscosity and/or mproved viscosity control in the dissolving pulp production process; and/or wherein the method results in increased reactivity of the dissolving pulp, preferably increased Fock's reactivity; and/or wherein the method results in increased content of oxidized groups of the dissolving pulp.

13. The method according to claim 1, further comprising a step of subjecting the dissolving pulp to a xylanase and/or a mannanase and/or a lipase and/or laccase and/or peroxidase.

14. A dissolving pulp made by the method according to claim 1.

15. A textile fiber or a derivatized cellulose made of the dissolving pulp according to claim 14.

16. (canceled)

17. The method according to claim 2, wherein the step of subjecting the dissolving pulp to a lytic polysaccharide monooxygenase and the step of subjecting the dissolving pulp to a cellulase are carried out sequentially.

18. The method according to claim 2 wherein the cellulase comprises SEQ ID NO; 4 of the mature polypeptide thereof, SEQ ID NO: 5 or the mature polypeptide thereof, SEQ ID NO: 6 or the mature polypeptide thereof, or SEQ ID NO: 7 or the mature polypeptide thereof.