U.S. patent application number 16/931149 was filed with the patent office on 2021-02-04 for methods and compositions for consumables.
The applicant listed for this patent is Impossible Foods Inc.. Invention is credited to Patrick O'Reilly Brown, Monte Casino.
Application Number | 20210030014 16/931149 |
Document ID | / |
Family ID | 1000005164053 |
Filed Date | 2021-02-04 |
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United States Patent
Application |
20210030014 |
Kind Code |
A1 |
Brown; Patrick O'Reilly ; et
al. |
February 4, 2021 |
Methods And Compositions For Consumables
Abstract
Provided herein are methods and compositions for the production
of cheese replicas. Generally the cheese replicas are produced by
inducing the enzymatic curdling of non-dairy milks.
Inventors: |
Brown; Patrick O'Reilly;
(Stanford, CA) ; Casino; Monte; (San Bruno,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Impossible Foods Inc. |
Redwood City |
CA |
US |
|
|
Family ID: |
1000005164053 |
Appl. No.: |
16/931149 |
Filed: |
July 16, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14666872 |
Mar 24, 2015 |
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16931149 |
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14152531 |
Jan 10, 2014 |
9011949 |
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14666872 |
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PCT/US2012/046552 |
Jul 12, 2012 |
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14152531 |
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61507096 |
Jul 12, 2011 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A23C 20/005 20130101;
A23C 11/106 20130101; A23K 20/147 20160501; A23C 20/025 20130101;
A23K 10/14 20160501; A23Y 2240/75 20130101; A23Y 2220/00 20130101;
A23Y 2280/00 20130101; A23K 50/48 20160501; A23C 20/02 20130101;
C12Y 203/02013 20130101 |
International
Class: |
A23C 20/02 20060101
A23C020/02; A23C 20/00 20060101 A23C020/00; A23K 10/14 20060101
A23K010/14; A23K 50/48 20060101 A23K050/48; A23K 20/147 20060101
A23K020/147; A23C 11/10 20060101 A23C011/10 |
Claims
1-30. (canceled)
31. A method of preparing a non-dairy milk composition comprising:
(a) providing an aqueous composition comprising one or more
isolated, enriched, or purified plant proteins, one or more oils or
fats, and a fermentable carbon source; (b) forming a colloidal
suspension, solution, or emulsion from the aqueous composition; and
(c) fermenting the colloidal suspension, solution, or emulsion
using one or more microbes selected from the group consisting of
Penicillium camemberti, Geotrichum candidum, Penicillium
roqueforti, Penicillium nalgiovensis, Verticillium lecanii,
Kluyveromyces lactis, Saccharomyces cerevisiae, Candida utilis,
Debaryomyces hansensil, Rhodosporidum infirmominiatum, Candida
jefer, Cornybacteria, Micrococcus sps., Lactobacillus sps.,
Lactococcus, Staphylococcus, Halomonas, Brevibacterium,
Psychrobacter, Leuconostocaceae, Streptococcus thermophilus,
Pediococcus sps., and Propionibacteria.
32. The method of claim 31, wherein the fermentable carbon source
is selected from the group consisting of a disaccharide, a fruit
juice, a monosaccharide, agave syrup, an oligosaccharide,
high-fructose corn syrup, honey, molasses, refined sugars,
sugar-cane extract, treacle, and mixtures thereof.
33. The method of claim 31, wherein the aqueous composition further
comprises an organic acid.
34. The method of claim 33, wherein the organic acid is selected
from the group consisting of acetic acid, citric acid, lactic acid,
malic acid, malonic acid, propionic acid, and mixtures thereof.
35. The method of claim 31, wherein the one or more isolated plant
proteins are selected from the group consisting of seed-storage
proteins from any seed, ribosomal proteins, actin, hexokinase,
lactate dehydrogenase, fructose bisphosphate aldolase,
phosphofructokinases, triose phosphate isomerases, phosphoglycerate
kinases, phosphoglycerate mutases, enolases, pyruvate kinases,
glyceraldehyde-3-phosphate dehydrogenases, pyruvate decarboxylases,
actins, translation elongation factors, ribulose-1,5-bisphosphate
carboxylase oxygenase (rubisco), ribulose-1,5-bisphosphate
carboxylase oxygenase activase (rubisco activase), albumins,
glycinins, conglycinins, globulins, vicilins, conalbumin, gliadin,
glutelin, gluten, glutenin, hordein, prolamin, phaseolin (protein),
proteinoplast, secalin, extensins, triticeae gluten, zein,
oleosins, caloleosins, steroleosins or other oil body proteins,
vegetative storage protein A, vegetative storage protein B, and
moong seed storage 8S globulin.
36. The method of claim 31, wherein the one or more oils or fats
are selected from the group consisting of corn oil, olive oil, soy
oil, peanut oil, walnut oil, almond oil, sesame oil, cottonseed
oil, rapeseed oil, canola oil, safflower oil, sunflower oil, flax
seed oil, algal oil, palm oil, palm kernel oil, coconut oil,
babassu oil, shea butter, mango butter, cocoa butter, wheat germ
oil, rice bran oil, oils produced by bacteria, algae, archaea or
fungi or genetically engineered bacteria, algae, archaea or fungi,
triglycerides, monoglycerides, diglycerides, sphingosides,
glycolipids, lecithin, lysolecithin, phophatidic acids,
lysophosphatidic acids, oleic acid, palmitoleic acid, palmitic
acid, myristic acid, lauric acid, myristoleic acid, caproic acid,
capric acid, caprylic acid, pelargonic acid, undecanoic acid,
linoleic acid, 20:1 eicosanoic acid, arachidonic acid,
eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated linoleic
acid, conjugated oleic acid, or esters of: oleic acid, palmitoleic
acid, palmitic acid, myristic acid, lauric acid, myristoleic acid,
caproic acid, capric acid, caprylic acid, pelargonic acid,
undecanoic acid, linoleic acid, 20:1 eicosanoic acid, arachidonic
acid, eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated
linoleic acid, or conjugated oleic acid, or glycerol esters of
oleic acid, palmitoleic acid, palmitic acid, myristic acid, lauric
acid, myristoleic acid, caproic acid, capric acid, caprylic acid,
pelargonic acid, undecanoic acid, linoleic acid, 20:1 eicosanoic
acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid,
18:2 conjugated linoleic acid, or conjugated oleic acid, and
triglyceride derivatives of oleic acid, palmitoleic acid, palmitic
acid, myristic acid, lauric acid, myristoleic acid, caproic acid,
capric acid, caprylic acid, pelargonic acid, undecanoic acid,
linoleic acid, 20:1 eicosanoic acid, arachidonic acid,
eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated linoleic
acid, and conjugated oleic acid.
37. The method of claim 31, wherein the aqueous composition further
comprises a starch.
38. The method of claim 37, wherein the starch is selected from the
group consisting of cornstarch, tapioca, wheat starch, and potato
starch.
39. The method of claim 31, further comprising pasteurizing or
sterilizing the non-dairy milk composition.
40. The method of claim 31, further comprising removing insoluble
solids from the aqueous composition.
41. The method of claim 40, wherein the insoluble solids are
removed from the aqueous compositing by centrifugation, straining,
filtering, settling, skimming, adding a coagulant, adding a
flocculant, or a combination thereof.
42. The method of claim 40, wherein removing the insoluble solids
from the aqueous composition results in a composition with less
than 50% of insoluble solids found in the aqueous composition
before the removal of the insoluble solids.
43. A non-dairy milk composition comprising: (a) a colloidal
suspension, solution, or emulsion comprising one or more isolated,
enriched, or purified plant proteins, and one or more oils or fats;
and (b) one or more microbes selected from the group consisting of
Penicillium camemberti, Geotrichum candidum, Penicillium
roqueforti, Penicillium nalgiovensis, Verticillium lecanii,
Kluyveromyces lactis, Saccharomyces cerevisiae, Candida utilis,
Debaryomyces hansensil, Rhodosporidum infirmominiatum, Candida
jefer, Cornybacteria, Micrococcus sps., Lactobacillus sps.,
Lactococcus, Staphylococcus, Halomonas, Brevibacterium,
Psychrobacter, Leuconostocaceae, Streptococcus thermophilus,
Pediococcus sps., and Propionibacteria.
44. The non-dairy milk composition of claim 43, wherein the aqueous
composition further comprises a fermentable carbon source.
45. The non-dairy milk composition of claim 44, wherein the
fermentable carbon source is selected from the group consisting of
a disaccharide, a fruit juice, a monosaccharide, agave syrup, an
oligosaccharide, high-fructose corn syrup, honey, molasses, refined
sugars, sugar-cane extract, treacle, and mixtures thereof.
46. The non-dairy milk composition of claim 43, wherein the aqueous
composition further comprises an organic acid.
47. The non-dairy milk composition of claim 46, wherein the organic
acid is selected from the group consisting of acetic acid, citric
acid, lactic acid, malic acid, malonic acid, propionic acid, and
mixtures thereof.
48. The non-dairy milk composition of claim 43, wherein the one or
more isolated plant proteins are selected from the group consisting
of seed-storage proteins from any seed, ribosomal proteins, actin,
hexokinase, lactate dehydrogenase, fructose bisphosphate aldolase,
phosphofructokinases, triose phosphate isomerases, phosphoglycerate
kinases, phosphoglycerate mutases, enolases, pyruvate kinases,
glyceraldehyde-3-phosphate dehydrogenases, pyruvate decarboxylases,
actins, translation elongation factors, ribulose-1,5-bisphosphate
carboxylase oxygenase (rubisco), ribulose-1,5-bisphosphate
carboxylase oxygenase activase (rubisco activase), albumins,
glycinins, conglycinins, globulins, vicilins, conalbumin, gliadin,
glutelin, gluten, glutenin, hordein, prolamin, phaseolin (protein),
proteinoplast, secalin, extensins, triticeae gluten, zein,
oleosins, caloleosins, steroleosins or other oil body proteins,
vegetative storage protein A, vegetative storage protein B, and
moong seed storage 8S globulin.
49. The non-dairy milk composition of claim 43, wherein the one or
more oils or fats are selected from the group consisting of corn
oil, olive oil, soy oil, peanut oil, walnut oil, almond oil, sesame
oil, cottonseed oil, rapeseed oil, canola oil, safflower oil,
sunflower oil, flax seed oil, algal oil, palm oil, palm kernel oil,
coconut oil, babassu oil, shea butter, mango butter, cocoa butter,
wheat germ oil, rice bran oil, oils produced by bacteria, algae,
archaea or fungi or genetically engineered bacteria, algae, archaea
or fungi, triglycerides, monoglycerides, diglycerides,
sphingosides, glycolipids, lecithin, lysolecithin, phophatidic
acids, lysophosphatidic acids, oleic acid, palmitoleic acid,
palmitic acid, myristic acid, lauric acid, myristoleic acid,
caproic acid, capric acid, caprylic acid, pelargonic acid,
undecanoic acid, linoleic acid, 20:1 eicosanoic acid, arachidonic
acid, eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated
linoleic acid, conjugated oleic acid, or esters of: oleic acid,
palmitoleic acid, palmitic acid, myristic acid, lauric acid,
myristoleic acid, caproic acid, capric acid, caprylic acid,
pelargonic acid, undecanoic acid, linoleic acid, 20:1 eicosanoic
acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid,
18:2 conjugated linoleic acid, or conjugated oleic acid, or
glycerol esters of oleic acid, palmitoleic acid, palmitic acid,
myristic acid, lauric acid, myristoleic acid, caproic acid, capric
acid, caprylic acid, pelargonic acid, undecanoic acid, linoleic
acid, 20:1 eicosanoic acid, arachidonic acid, eicosapentanoic acid,
docosohexanoic acid, 18:2 conjugated linoleic acid, or conjugated
oleic acid, and triglyceride derivatives of oleic acid, palmitoleic
acid, palmitic acid, myristic acid, lauric acid, myristoleic acid,
caproic acid, capric acid, caprylic acid, pelargonic acid,
undecanoic acid, linoleic acid, 20:1 eicosanoic acid, arachidonic
acid, eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated
linoleic acid, and conjugated oleic acid.
50. The non-dairy milk composition of claim 43, wherein the aqueous
composition further comprises a starch.
51. The non-dairy milk composition of claim 50, wherein the starch
is selected from the group consisting of cornstarch, tapioca, wheat
starch, and potato starch.
52. The non-dairy milk composition of claim 43, wherein the
non-dairy milk composition has at least 50% of its insoluble solids
removed.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 14/666,872 filed Mar. 24, 2015 which is a
continuation of U.S. patent application Ser. No. 14/152,531 filed
Jan. 10, 2014, which is a continuation of PCT/US2012/046552, filed
Jul. 12, 2012, which claims priority to U.S. Application Ser. No.
61/507,096, filed Jul. 12, 2011, the disclosures of which are
incorporated herein by reference in their entirety.
BACKGROUND OF THE INVENTION
[0002] Cheese making has relied on dairy milks as the major
ingredient for more than 4000 years. Dairy cheese is made from
curds formed from dairy milk. Dairy milks can readily be made to
form curds suitable for making cheese by contacting the dairy milk
with rennet (an aspartic protease which cleaves kappa-casein) at
mildly acidic pH. Some cheeses, e.g., cream cheese, ricotta,
cottage cheese and paneer, are made without rennet. In the absence
of rennet, dairy cheese may be induced to curdle with acid (e.g.,
lemon juice, vinegar, etc.) or a combination of heat and acid. Acid
coagulation can also occur naturally from starter culture
fermentation. The strength of the curds depends on the type of
coagulation. Most commercially produced cheeses use some type of
rennet (animal, vegetable or microbial-derived) in their
production.
[0003] The global dairy sector contributes an estimated 4 percent
to the total global anthropogenic greenhouse gas emissions.
Producing 1 kg of cheddar cheese requires an average of 10,000
Liters of fresh water. Additionally, many individuals cannot digest
and metabolize lactose. In these individuals enteric bacteria
ferment the lactose, resulting in various abdominal symptoms, which
can include abdominal pain, bloating, flatulence, diarrhea, nausea,
and acid reflux. Additionally, the presence of lactose and its
fermentation products raises the osmotic pressure of the colon
contents. 3.4% of children in the U.S.A. are reported to have
allergies to dairy milks. Many individuals choose to avoid milk for
ethical or religious reasons.
[0004] Non-dairy milks, including plant-derived milks avoid many of
the environmental, food sensitivity, ethical and religious problems
associated with dairy milk and they can be made free of lactose,
making the generation of dairy substitutes using the plant derived
milks attractive. However, rennet is not an effective agent for
inducing non-dairy proteins or emulsions, including plant-derived
milks, including almond milk, chestnut milk, pecan milk, hazelnut
milk, cashew milk, pine nut milk, and walnut milk, to curdle.
Consequently, traditional cheese making techniques have not been
successfully used to produce non-dairy cheese replicas.
[0005] Flavor and aroma in dairy cheese results in part from the
degradation of lactose, proteins and fats, carried out by ripening
agents, which include: bacteria and enzymes in the milk, bacterial
cultures added during the cheese-making process, rennet, other
proteases, lipases, added molds and/or yeasts and bacteria and
fungi that opportunistically colonize the cheese during ripening
and aging.
[0006] Cheese replicas made principally of non-dairy ingredients
are commercially available. Most of these cheese replicas include
some dairy ingredients, for example, casein. Some commercially
available cheese replicas contain no animal products. These include
fermented cheese replicas made from nut milks from which insoluble
carbohydrates have not been effectively removed, and made without
using a protein crosslinking agent and several products in which a
starch is a principal ingredient or containing agar, carrageenan
and tofu to provide the desired texture. Most tasters consider none
of the currently available cheese replicas to adequately replicate
the taste, aroma and mouthfeel of dairy cheeses.
[0007] Complex carbohydrates in currently available cheese replicas
made from nut milks have unfavorable effects on the texture,
resulting in a product with a grainy mouthfeel and lacking the
creaminess of dairy cheeses.
[0008] Starches that comprise the major gelling agent in many
currently available cheese replicas lead to a relatively high
carbohydrate content, which may be undesirable to consumers, for
example those wishing to limit carbohydrate intake.
[0009] Because of these deficiencies, there is currently no cheese
replica that is acceptable to most consumers as an alternative to
traditional dairy cheeses.
[0010] Thus, it is clear that there is a great need in the art for
an improved method and system for producing non-dairy cheese
replicas while avoiding the shortcomings and drawbacks of the
cheese replicas that have previously been available to
consumers.
SUMMARY OF THE INVENTION
[0011] The invention relates to methods and compositions for
non-dairy milk and cheese products, including without limitation,
plant-derived milk and cheese products, as an alternative to dairy
products for human consumption.
[0012] The invention provides a method for producing a non-dairy
cheese by preparing an emulsion comprising proteins and fats from
plants or other non-animal sources, inducing the emulsion to form a
gel by enzymatic cross-linking of the proteins or denaturing the
proteins, and producing a cheese-replica from the gel. In one
embodiment, the emulsion contains less than 10% animal products. In
one embodiment, the emulsion contains less than 8%, 7%, 6%, 5%, or
3% animal products. In one embodiment the emulsion contains no
animal products. In one embodiment the inducing step of said method
comprises adding an enzyme. In one embodiment of said method the
enzyme used is transglutaminase. In one embodiment the enzyme used
is Factor XIII (fibrin-stabilizing factor). In one embodiment of
said method the enzyme used is Keratinocyte transglutaminase
(TGM1). In one embodiment the enzyme used is Tissue
transglutaminase (TGM2). In one embodiment of said method the
enzyme used is Epidermal transglutaminase (TGM3). In one embodiment
the enzyme used is Prostate transglutaminase (TGM4). In one
embodiment the enzyme used is TGM X (TGM5). In one embodiment of
said method the enzyme used is TGM Y (TGM6). In one embodiment of
said method the enzyme used is TGM Z (TGM7). In one embodiment, the
enzyme is a transglutaminase from Streptoverticillium mobaraense.
In one embodiment, the enzyme is similar or identical to a
transglutaminase from Streptoverticillium mobaraense. In one
embodiment, the enzyme is a lysyl oxidase. In one embodiment the
emulsion is non-dairy milk. In one embodiment at least one of the
plant sources are nuts. In one embodiment at least one of the plant
sources are legumes. In one embodiment at least one of the plant
sources are seeds. In one embodiment at least one of the plant
sources are leaves. In one embodiment at least one of the plant
sources are fruit from the family Fabaceae. In one embodiment at
least one of the non-animal sources is a species of bacteria. In
one embodiment at least one of the non-animal sources is a species
of archaea. In one embodiment at least one of the non-animal
sources is a species of fungus. In one embodiment at least one of
the non-animal sources is a species of algae. In one embodiment the
emulsion is a non-dairy milk. In one embodiment the nuts are one or
more of following: almonds, cashews, brazilnuts, chestnuts,
coconuts, hazelnuts, macadamia nuts, peanuts, pecans, pistachios or
walnuts. One embodiment of said method has an additional step of
adding a sugar to the emulsion. In one embodiment the sugar added
to the emulsion is a monosaccharide. In one embodiment the sugar
added to the emulsion is a disaccharide. In one embodiment said
method has an additional step of inoculating with lactic acid
bacteria. In the embodiment of inoculating with lactic acid
bacteria the invention provides an addition step of allowing growth
of the bacterial cultures. In one embodiment the method can include
an additional step of adding one or more of the following
Penicillium camemberti, Geotrichum candidum, Penicillium
roqueforti, Penicillium nalgiovensis, Verticillium lecanii,
Kluyveromyces lactis, Saccharomyces cerevisiae, Candida utilis,
Debaryomyces hansensil, Rhodosporidum infirmominiatum, Candida
jefer, Cornybacteria, Micrococcus sps., Lactobacillus sps.,
Lactococcus, Staphylococcus, Halomonas, Brevibacterium,
Psychrobacter, Leuconostocaceae, Streptococcus thermophilus,
Pediococcus sps., or Propionibacteria. In one embodiment the method
can include an additional step of adding one or more of the
following halophilc bacterium or archaea. In one embodiment the
invention provides for the use of thermal denaturing and
cross-linking. In one embodiment the invention provides thermal
denaturing used without the addition of any enzymes. One embodiment
of said method has an additional step of cutting the gel. One
embodiment of said method has an additional step of draining and
shaping the cut gel. One embodiment of said method has an
additional step of adding a flavoring component. In one embodiment
the flavoring component is one or more species of bacteria. In one
embodiment the flavoring component is lactic acid bacteria. In one
embodiment the flavoring component is a mold. In one embodiment the
flavoring component is yeast. In one embodiment the method includes
concentrated proteins prior to preparing the emulsion. In one
embodiment the concentrated proteins are each essentially
homogeneous proteins. In one embodiment the method includes the
additional step of concentrating proteins and fats from one or more
plant sources. In one embodiment the method includes the additional
step of concentrating proteins and fats from one or more non-animal
sources. In one embodiment the method includes the additional step
of isolating the proteins and fats from one or more plant sources.
In one embodiment the method includes the additional step of
isolating the proteins and fats from one or more non-animal
sources. In one embodiment the method includes the additional step
of purifying the proteins and fats from one or more plant sources.
In one embodiment the method includes the additional step of
purifying the proteins and fats from one or more non-animal
sources. In one embodiment the invention provides for the
concentrated proteins to come from the same essentially homogeneous
plant-derived protein. In one embodiment the invention provides for
the concentrated proteins to come from the same essentially
homogeneous non-animal-derived protein. In one embodiment the
invention provides for the concentrated proteins to come from
separate plant species. In one embodiment the invention provides
for the concentrated proteins to come from separate non-animal
species.
[0013] In addition, the invention provides steps for preparation to
undertake the claimed methods of producing a non-animal based
cheese including (a) obtaining nuts or seeds, and (b) surface
sterilizing the nuts or seeds. In one embodiment the method uses
nuts or seeds as the plant sources. In one embodiment the surface
sterilization step is blanching procedure. In one embodiment the
preparation method can also include a step for washing the nuts or
seeds. In one embodiment the method of preparation can also include
the step of decompounding the nuts or seeds. In one embodiment the
step of decompounding the nuts or seeds is a blending procedure. In
one embodiment the method of preparation can also include
centrifugation. In one embodiment the method of preparation can
also include a centrifugation procedure that results in the removal
of at least 85% of the suspended solids. In one embodiment the
method of preparation can also include a centrifugation procedure
that results in the removal of at least 75%, 65%, 55%, or 45% of
the suspended solids.
[0014] The invention provides a method comprising decompounding
nuts or seeds in water, removing at least 85% of the suspended
solids and adding a transglutaminase to catalyze the formation of
crosslinks between proteins from the nuts or seeds.
[0015] The invention provides a method for making a non-dairy
cheese replica comprising, obtaining nut milk, centrifugation of
the nut milk to remove insoluble matter, and crosslinking proteins
within the nut milk with transglutaminase.
[0016] In one embodiment the invention provides a composition
comprising centrifuged non-dairy milk wherein at least 85% of the
insoluble solids have been removed relative to the non-dairy milk
prior to centrifugation.
[0017] In one embodiment the invention provides a composition
comprising a non-dairy milk with less than 30% polysaccharides. In
one embodiment the non-dairy milk composition has less than 10%
polysaccharides. In one embodiment the non-dairy milk composition
less than 1% polysaccharides. In one embodiment the non-dairy milk
composition is entirely composed of ingredients derived from
non-animal sources. In one embodiment the non-dairy milk
composition is at least comprised of 20% protein isolated from a
single plant species.
[0018] In one embodiment the invention provides for a composition
comprised of non-dairy milk and a cross-linking enzyme. In one
embodiment of the non-dairy milk and a cross-linking enzyme
composition the cross-linking enzyme is Factor XIII
(fibrin-stabilizing factor). In one embodiment of the non-dairy
milk and a cross-linking enzyme composition, the enzyme is a
transglutaminase from Streptoverticillium mobaraense. In one
embodiment of the non-dairy milk and a cross-linking enzyme
composition, the enzyme is similar or identical to a
transglutaminase from Streptoverticillium mobaraense. In one
embodiment of the non-dairy milk and a cross-linking enzyme
composition the cross-linking enzyme is Keratinocyte
transglutaminase (TGM1). In one embodiment of the non-dairy milk
and a cross-linking enzyme composition the cross-linking enzyme is
Tissue transglutaminase (TGM2). In one embodiment of the non-dairy
milk and a cross-linking enzyme composition the cross-linking
enzyme is Epidermal transglutaminase (TGM3). In one embodiment of
the non-dairy milk and a cross-linking enzyme composition the
cross-linking enzyme is Prostate transglutaminase (TGM4). In one
embodiment of the non-dairy milk and a cross-linking enzyme
composition the cross-linking enzyme is TGM X (TGM5). In one
embodiment of the non-dairy milk and a cross-linking enzyme
composition the cross-linking enzyme is TGM Y (TGM6). In one
embodiment of the non-dairy milk and a cross-linking enzyme
composition the cross-linking enzyme is TGM Z (TGM7). In one
embodiment the invention provides for a composition comprised of
non-dairy milk and a cross-linking enzyme. In one embodiment of the
non-dairy milk and a cross-linking enzyme composition the
cross-linking enzyme is lysyl oxidase. In one embodiment of the
composition comprising non-dairy milk and a cross-linking enzyme,
protein crosslinks are formed between glutamine and lysine side
chains of respective protein constituents.
[0019] In one embodiment of the non-dairy milk and a cross-linking
enzyme composition, at least 85% of the insoluble solids have been
removed by centrifugation. In one embodiment of the non-dairy milk
and a cross-linking enzyme composition, proteins from non-animal
sources comprise at least 50% of the total mass of the composition.
In one embodiment of the non-dairy milk and a cross-linking enzyme
composition the starch content is less than 1% by mass. In one
embodiment of the non-dairy milk and a cross-linking enzyme
composition is the starch content is less than 5% by mass. One
embodiment of the non-dairy milk and a cross-linking enzyme
composition has less than 1% insoluble carbohydrate by mass. One
embodiment the non-dairy milk and a cross-linking enzyme
composition has less than 5% insoluble carbohydrate by mass. One
embodiment the non-dairy milk and a cross-linking enzyme
composition has less than 5% starch and less than 1% insoluble
carbohydrate by mass. One embodiment the non-dairy milk and a
cross-linking enzyme composition has less than 5% starch and less
than 5% insoluble carbohydrate by mass. In one embodiment the
non-dairy milk and a cross-linking enzyme composition has a
polysaccharide content less than 1% by mass. In one embodiment the
non-dairy milk and a cross-linking enzyme composition has a
polysaccharide content less than 5% by mass. In one embodiment the
non-dairy milk and a cross-linking enzyme composition has a
carbohydrate content less than 1% by mass. In one embodiment the
non-dairy milk and a cross-linking enzyme composition has a
carbohydrate content less than 5% by mass. In one embodiment the
non-dairy milk and a cross-linking enzyme composition has a
carbohydrate content less than 10% by mass. In one embodiment of
the non-dairy milk and a cross-linking enzyme composition, at least
80% of the protein content comprises a single monomeric or
multimeric protein.
[0020] In some embodiments the invention provides for a composition
comprising covalently crosslinked non-dairy proteins, a
crosslinking enzyme, and cheese microbes. In one embodiment of the
composition of covalently crosslinked non-dairy proteins, a
crosslinking enzyme, and cheese microbes composition, the proteins
are not from soy. In one embodiment of the composition comprising
covalently crosslinked non-dairy proteins, a crosslinking enzyme,
and cheese microbes, less than 5% of the proteins are from soy. In
one embodiment the composition comprises less than 20% insoluble
solids. In one embodiment the covalently crosslinked non-dairy
proteins, a crosslinking enzyme, and cheese microbes composition
also contains oils or fats isolated from non-dairy sources. In one
embodiment of the composition comprising covalently crosslinked
non-dairy proteins, a crosslinking enzyme, and cheese microbes the
crosslinking enzyme of the composition is a transglutaminase. In
one embodiment of the composition comprising covalently crosslinked
non-dairy proteins, a crosslinking enzyme, and cheese microbes the
crosslinking enzyme of the composition is a lysyl oxidase. In one
embodiment of the composition comprising covalently crosslinked
non-dairy proteins, a crosslinking enzyme, and cheese microbes the
cross-linking enzyme is Factor XIII (fibrin-stabilizing factor),
Keratinocyte transglutaminase (TGM1), Tissue transglutaminase
(TGM2), Epidermal transglutaminase (TGM3), Prostate
transglutaminase (TGM4), TGM X (TGM5), TGM Y (TGM6), TGM Z (TGM7.
In one embodiment of the composition comprising non-dairy milk and
a cross-linking enzyme and cheese microbes, the enzyme is a
transglutaminase from Streptoverticillium mobaraense. In one
embodiment of the composition comprising non-dairy milk and a
cross-linking enzyme and cheese microbes, the enzyme is similar or
identical to a transglutaminase from Streptoverticillium
mobaraense. In one embodiment of the composition comprising
non-dairy milk and a cross-linking enzyme and cheese microbes,
protein crosslinks are formed between glutamine and lysine side
chains of respective protein constituents. In one embodiment of the
composition comprising non-dairy milk and a cross-linking enzyme
and cheese microbes, protein crosslinks are formed between lysine
side chains of respective protein constituents.
[0021] In some embodiments the invention provides for a cheese
replica composition comprised of a gelled emulsion of non-dairy
proteins and fats. In one embodiment the cheese replica composition
has between 10% and 40% proteins from non-dairy sources and between
0% and 65% fats from non-dairy sources. In one embodiment the
cheese replica composition further comprises a crosslinking enzyme
selected from the group consisting of Factor XIII
(fibrin-stabilizing factor), Keratinocyte transglutaminase (TGM1),
Tissue transglutaminase (TGM2), Epidermal transglutaminase (TGM3),
Prostate transglutaminase (TGM4), TGM X (TGM5), TGM Y (TGM6) or TGM
Z (TGM7). In one embodiment of the cheese replica composition, the
enzyme is a transglutaminase from Streptoverticillium mobaraense.
In one embodiment of the cheese replica composition, the enzyme is
similar or identical to a transglutaminase from Streptoverticillium
mobaraense. In one embodiment of the cheese replica composition,
protein crosslinks are formed between glutamine and lysine side
chains of respective protein constituents. In one embodiment of the
cheese replica composition, protein crosslinks are formed between
lysine side chains of respective protein constituents. In some
embodiment of the cheese replica composition, formation of protein
crosslinks is catalyzed by a lysyl oxidase.
[0022] In one embodiment the cheese replica composition further
comprises cheese-making microbes. In one embodiment the cheese
replica composition has one or more cheese-making microbes from the
group consisting of Penicillium camemberti, Geotrichum candidum,
Penicillium roqueforti, Penicillium nalgiovensis, Verticillium
lecanii, Kluyveromyces lactis, Saccharomyces cerevisiae, Candida
utilis, Debaryomyces hansensil, Rhodosporidum infirmominiatum,
Candida jefer, Cornybacteria, Micrococcus sps., Lactobacillus sps.,
Lactococcus, Staphylococcus, Halomonas, Brevibacterium,
Psychrobacter, Leuconostocaceae, Streptococcus thermophilus,
Pediococcus sps., or Propionibacteria.
[0023] In some embodiments the invention provides for a cheese
replica composition, wherein the cheese replica is a cheese
equivalent according to a human. In some embodiments the invention
provides for a cheese replica composition, wherein human subjects
cannot distinguish the cheese replica from a dairy cheese. In one
embodiment of the cheese replica at least 20% of the protein
content comprises a single monomeric or multimeric protein. In one
embodiment the cheese replica composition is made without the
addition of starches or rennet or any other extrinsic proteases
other than those contributed by microbial cultures. In one
embodiment the cheese replica composition has a pH less than 5.5
where the acidification was accomplished solely by microbial
fermentation. In one embodiment the cheese replica composition has
a pH less than 5 where the acidification was accomplished solely by
microbial fermentation. In one embodiment the cheese replica
composition has a pH less than 6 where the acidification was
accomplished solely by microbial. In one embodiment the cheese
replica composition is contains no animal products. In one
embodiment the cheese replica has less than 5% by mass insoluble
carbohydrates. In one embodiment the cheese replica composition
contains no rennet, vinegar, or lemon juice.
[0024] In some embodiments the invention provides for a cheese
replica composition made with the addition of rennet or any
aspartic protease or any other type protease (e.g., serine
protease) to affect or enhance the flavor and/or aroma and/or
texture of the cheese replica.
[0025] In some embodiments the invention provides for a cheese
replica composition made with the addition of vinegar, lemon juice
or any other type of acid to affect or enhance the flavor and/or
aroma and/or texture of the cheese replica.
[0026] In some embodiments the invention provides for a soft fresh
cheese replica composition composed of pasteurized almond milk,
pasteurized macadamia nut milk, mesophilic starter culture,
transglutaminase, water, and salt. In one embodiment the soft fresh
cheese replica composition includes the addition of vinegar. In one
embodiment the soft fresh cheese replica composition includes the
addition of microbial coagulant. In one embodiment the soft fresh
cheese replica composition includes the addition of vinegar and
microbial coagulant.
[0027] In some embodiments the invention provides for a salted
cheese replica composition composed of pasteurized almond milk,
pasteurized macadamia nut milk, mesophilic starter culture,
transglutaminase, and water. In one embodiment the salted cheese
replica composition includes the addition of vinegar. In one
embodiment the salted cheese replica composition includes a
microbial coagulant. In one embodiment the salted cheese replica
composition includes vinegar and a microbial coagulant.
[0028] In some embodiments the invention provides for a soft
ripened cheese replica composition composed of pasteurized almond
milk, pasteurized macadamia nut milk, mesophilic starter culture,
flora danica, Geotrichum candidum, Penicillium candidum,
Debaromyces hansenii, transglutaminase, water, and salt. In one
embodiment the soft ripened cheese replica composition includes
vinegar. In one embodiment the soft ripened cheese replica
composition includes a microbial coagulant. In one embodiment the
soft ripened cheese replica composition includes vinegar and
microbial coagulant.
[0029] In some embodiments the invention provides for a goat cheese
replica composition composed of pasteurized almond milk, macadamia
nut milk, mesophilic starter culture, transglutaminase, water, and
salt. In one embodiment the goat cheese replica composition
includes vinegar. In one embodiment the goat cheese replica
composition includes a microbial coagulant. In one embodiment the
goat cheese replica composition includes vinegar and a microbial
coagulant.
[0030] In some embodiments the invention provides a method for
making a cheese replica comprising removing from a non-dairy milk a
proportion of insoluble solids. In some embodiments the invention
provides a method for making a cheese replica comprising removing
from a plant-based milk a proportion of insoluble solids. In some
embodiments the invention provides a method comprising centrifuging
a non-dairy milk to sediment a proportion of insoluble solids. In
some embodiments about 10%, about 20%, about 30%, about 40%, about
50%, about 60%, about 70%, about 80%, about 90%, or about 100% of
the insoluble solids are removed.
[0031] In some embodiments the isolated or enriched proteins of the
methods and compositions of the invention comprise one or more of:
ribosomal proteins, actin, hexokinase, lactate dehydrogenase,
fructose bisphosphate aldolase, phosphofructokinases, triose
phosphate isomerases, phosphoglycerate kinases, phosphoglycerate
mutases, enolases, pyruvate kinases, glyceraldehyde-3-phosphate
dehydrogenases, pyruvate decarboxylases, actins, translation
elongation factors, ribulose-1,5-bisphosphate carboxylase oxygenase
(rubisco), ribulose-1,5-bisphosphate carboxylase oxygenase activase
(rubisco activase), albumins, glycinins, conglycinins, globulins,
vicilins, conalbumin, gliadin, glutelin, gluten, glutenin, hordein,
prolamin, phaseolin (protein), proteinoplast, secalin, extensins,
triticeae gluten, zein, any seed storage protein, oleosins,
caloleosins, steroleosins or other oil body proteins, vegetative
storage protein A, vegetative storage protein B, moong seed storage
8S globulin.
[0032] In some embodiments the invention provides a composition
comprising (A) covalently crosslinked plant-derived proteins from a
set comprising ribosomal proteins, actin, hexokinase, lactate
dehydrogenase, fructose bisphosphate aldolase,
phosphofructokinases, triose phosphate isomerases, phosphoglycerate
kinases, phosphoglycerate mutases, enolases, pyruvate kinases,
glyceraldehyde-3-phosphate dehydrogenases, pyruvate decarboxylases,
actins, translation elongation factors, ribulose-1,5-bisphosphate
carboxylase oxygenase (rubisco), ribulose-1,5-bisphosphate
carboxylase oxygenase activase (rubisco activase), albumins,
glycinins, conglycinins, globulins, vicilins, conalbumin, gliadin,
glutelin, gluten, glutenin, hordein, prolamin, phaseolin (protein),
proteinoplast, secalin, extensins, triticeae gluten, zein, any seed
storage protein, oleosins, caloleosins, steroleosins or other oil
body proteins, vegetative storage protein A, vegetative storage
protein B, and moong seed storage 8S globulin; (B) a crosslinking
enzyme, and (C) cheese microbes.
[0033] In some embodiments the invention provides a composition
comprising (A) covalently crosslinked plant-derived proteins from a
set comprising ribosomal proteins, actin, hexokinase, lactate
dehydrogenase, fructose bisphosphate aldolase,
phosphofructokinases, triose phosphate isomerases, phosphoglycerate
kinases, phosphoglycerate mutases, enolases, pyruvate kinases,
glyceraldehyde-3-phosphate dehydrogenases, pyruvate decarboxylases,
actins, translation elongation factors, ribulose-1,5-bisphosphate
carboxylase oxygenase (rubisco), ribulose-1,5-bisphosphate
carboxylase oxygenase activase (rubisco activase), albumins,
glycinins, conglycinins, globulins, vicilins, conalbumin, gliadin,
glutelin, gluten, glutenin, hordein, prolamin, phaseolin (protein),
proteinoplast, secalin, extensins, triticeae gluten, zein, any seed
storage protein, oleosins, caloleosins, steroleosins or other oil
body proteins, vegetative storage protein A, vegetative storage
protein B, and moong seed storage 8S globulin.
INCORPORATION BY REFERENCE
[0034] All publications and patent applications mentioned in this
specification are herein incorporated by reference to the same
extent as if each individual publication or patent application was
specifically and individually indicated to be incorporated by
reference.
BRIEF DESCRIPTION OF THE DRAWINGS
[0035] The novel features of the invention are set forth with
particularity in the appended claims. A better understanding of the
features and advantages of the present invention will be obtained
by reference to the following detailed description that sets forth
illustrative embodiments, in which the principles of the invention
are utilized, and the accompanying drawings of which:
[0036] FIG. 1 shows freshly cut curds prepared by the method used
to produce the soft fresh cheese replicas and salted cheese
replicas described in the examples.
[0037] FIG. 2 shows drained curds prepared by the method used to
produce the soft fresh cheese replicas and salted cheese replicas
described in the examples.
[0038] FIG. 3 shows soft fresh cheese replicas made from non-dairy
milk.
[0039] FIG. 4 shows Annatto-colored cheese replicas made from
non-dairy milk.
[0040] FIG. 5 shows salted cheese replica made from non-dairy
milk.
[0041] FIG. 6 shows salted cheese replica with paprika and fennel
pollen made from non-dairy milk.
[0042] FIG. 7 shows waxed cheese replica made from non-dairy
milk.
[0043] FIG. 8 shows soft ripened cheese replica made from non-dairy
milk
[0044] FIG. 9 shows goat cheese replicas made from non-dairy milk
(A) with black pepper (B) with chives.
[0045] FIG. 10 shows cheese replica gels made from partially
purified plant proteins and vegetable oil, cultured with mesophilic
culture MA11 (Danisco): (A) depicts cultured cheese replicas made
from 2% Pea albumin fraction, 3% vegetable oil, heated and
crosslinked with transglutaminase; (B) depicts cultured cheese
replicas made from 2% Moong bean 8S globulin fraction, 3% vegetable
oil, crosslinked with transglutaminase, without prior heating; (C)
depicts cultured cheese replicas made from 7.5% Pea globulin
fraction, 3% vegetable oil, heated and crosslinked with
transglutaminase; (D) depicts cultured cheese replicas made from
7.5% Moong bean 8S globulin fraction, 3% vegetable oil, crosslinked
with transglutaminase, without prior heating of proteins.
DETAILED DESCRIPTION OF THE INVENTION
[0046] The present invention discloses methods and compositions
based upon the realization that transglutaminase efficiently
crosslinks the proteins in many non-dairy milks to produce a soft,
moist, elastic gel (a "curdling" of non-dairy milks). This process
allows the separation of the crosslinked proteins and associated
fats from the "whey." The crosslinked proteins can hold a fat
emulsion, and have the essential physical characteristics needed
for pressing, culturing and ripening a cheese replica derived from
non-dairy milk. In various embodiments the current invention
includes cheese replicas principally, entirely or partially
composed of ingredients derived from non-animal sources. In
additional embodiments the present invention includes methods for
making cheese replicas from non-animal sources. In various
embodiments these results are achieved by replicating the curdling
process of cheese making in non-dairy milks using enzymes.
[0047] The term a cheese "substitute" or "replica" can be any
non-dairy product that serves a role as food or in food that is
commonly served by traditional dairy cheese. A cheese "substitute"
or "replica" can be a product that shares visual, olfactory,
textural or taste characteristics of cheese such that an ordinary
human observer of the product is induced to think of traditional
dairy cheese.
[0048] A purified protein can be a preparation in which the
cumulative abundance by mass of protein components other than the
specified protein, which can be a single monomeric or multimeric
protein species, is reduced by a factor of 3 or more, or a factor
of 5 or more, or a factor of 10 or more relative to the source
material from which the specified protein was said to be
purified.
[0049] The term "homogeneous" can mean a single protein component
comprises more than 90% by mass of the total protein constituents
of a preparation.
[0050] The term resemble can mean one composition having
characteristics recognizably similar to another composition by an
ordinary human observer.
[0051] The term "indistinguishable" can mean that an ordinary human
observer would not be able to differentiate two compositions based
on one or more characteristics. It is possible that two
compositions are indistinguishable based on one characteristic but
not based on another, for example two compositions can have
indistinguishable taste while having colors that are different.
Indistinguishable can also mean that the product provides an
equivalent function as or performs an equivalent role as the
product for which it is substituting.
I. Non-Dairy Milks
[0052] Cheese replicas may be made using non-dairy milk prepared
from nuts or plant seeds. For example almonds, cashews, brazilnuts,
chestnuts, coconuts, hazelnuts, macadamia nuts, peanuts, pecans,
pistachios or walnuts can provide non-dairy milk that, in various
embodiments of the invention, is used to produce cheese replicas.
Nuts can include so called "true nuts" as well as a wide variety of
dried seeds from of plants. Any large, oily kernel found within a
shell and used in food may be regarded as a nut. Plant seeds can
include a wide variety embryonic plants enclosed in a seed coat.
Plant seeds can include for example legumes, cereals, and
gymnosperms. Non-dairy products or compositions include products or
compositions where the constituent proteins, fats and/or small
molecules can be isolated from or secreted by plants, bacteria,
viruses, archaea, fungi, algae, or they can be made synthetically
by chemical synthesis or in vitro. Non-dairy products are generally
not derived from cows, goats, buffalo, sheep, horses, camels, or
other mammals. In some embodiments non-dairy products do not
contain dairy proteins. In some embodiments non-dairy products do
not contain dairy fats. In some embodiments non-dairy products do
not contain enzymes derived from an animal.
[0053] The nuts or seeds of the present invention can be raw. In
some embodiments the nuts or seeds used to produce non-dairy milk
are all raw. Alternatively some or all of the nuts or seeds used in
the production of a non-dairy milk can be processed. The processed
nuts or seed can be roasted, dry roasted, toasted, or baked.
[0054] Milk, or non-dairy milk, can mean an emulsion comprising
proteins and fats or a solution or suspension of proteins,
sometimes further comprising other solutes that might include
carbohydrates, salts and other small molecules that contribute to
flavor, texture, emulsion stability, protein solubility or
suspension stability, or its ability to support growth of microbial
cultures used in making cheese replicas, yogurt replicas, or other
replicas of cultured dairy products.
[0055] The non-dairy milk can be made by a method comprising
preparing the nuts or plant seeds with processing steps such as
sterilizing, blanching, shocking, decompounding, centrifugation, or
washing.
[0056] The non-dairy milks can be produced by decompounding the
nuts or plant seeds, for example by grinding or blending or milling
the nuts in a solution comprising water. In various embodiments
alternative methods for decompounding the nuts or dried seeds
include crushing, tumbling, crumbling, atomizing, shaving,
pulverizing, grinding, milling, water eroding (for example with a
water jet), or finely chopping the nuts or plant seeds. In some
embodiments the decompounding step takes place in a blender. In
some embodiments the decompounding is in a continuous flow grinder.
In some embodiments the decompounding is in a continuous flow mill.
The decompounding can be followed by a sorting, filtering,
screening, or separation step. In some embodiments the decompounded
nuts or seeds can be stored prior to the formation of a non-dairy
milk. In some embodiments an aqueous solution is added before,
during, or after the decompounding.
[0057] The nuts or seeds used in some embodiments of the invention
to make non-dairy milks may have contaminants on the surface which
would make a non-dairy milk unsafe or unpalatable. Accordingly the
nuts or seeds can be washed prior to use. The nuts or seeds can
also be sterilized to remove, reduce, or kill any contaminants on
the surface of the nuts or seeds. A sterilization step can be an
irradiation step, a heat step (e.g. steam sterilization, flaming,
or dry heat), or a chemical sterilization (e.g, exposure to ozone).
In some embodiments the sterilization step kills more than 95% of
microbes on the nuts or seeds. In some embodiments the
sterilization step kills more than 99% of microbes on the nuts or
seeds. Blanching is a process wherein the food is exposed to hot
water (e.g. boiling), removed after a brief timed interval, and
finally cooled by exposure to cold water (e.g. iced or cold running
water). When nuts, such as almonds or pistachios, are blanched, the
skin of the nut (botanically the seed coat surrounding the embryo)
softens and can be easily removed later. Accordingly, in some
embodiments the invention provides for non-dairy milk compositions
with a reduced percentage of components found in the skin of the
nut or seed. For example the composition can have 50%, 40%, 30%,
20%, 10% or less than 5% of the seed coat remaining after the
preparation process. In some embodiments the invention provides for
methods for making a non-dairy milk comprising removing the seed
coat.
[0058] In one embodiment the blanching procedure is as follows:
Place nuts into 212.degree. F. heated water and blanch for 30
seconds. Drain nuts. Immediately move drained nuts into cold water.
In some embodiments the temperature of the water is altered by 5%,
10% or 20%. In some embodiments the cold water temperature is about
0.degree. C., 5.degree. C., 10.degree. C., or 20.degree. C. In some
embodiments the blanching takes about 10, 20, 30, 40, 50 seconds or
about 1, 2, or 5 minutes.
[0059] In some embodiments the nuts or seeds can be hydrated, for
example by immersion in water for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 hours. In
some embodiments the hydration step lasts multiple days. In some
embodiments the water contains other components such as salts or
preservatives. In some embodiments the water is maintained at a
constant cool temperature. In some embodiments the hydration step
can occur prior to the decompounding step.
[0060] In some embodiments the nuts or seeds can be dried, for
example by exposure to a low humidity and/or heated environment for
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,
20, 21, 22, 23, or 24 hours. In some embodiments the drying step
lasts multiple days. The drying step can occur prior to the
decompounding step or the drying step can occur after a
decompounding step.
[0061] In one embodiment the decompounding step is a blending
procedure as follows: Place nuts in a clean and sanitized blender.
Add clean fresh water to the blender. Turn on the blender and
gradually increase speed up to maximum, blending for 5 minutes.
Accumulate blended slurry in clean and sanitized ice cold
container. Immediately begin rapid cooling of slurry by stirring
slurry. Once the slurry has cooled to 50.degree. F., move slurry to
refrigerator to continue cooling to below 41.degree. F. Allow
slurry to rest at 36.degree. F. overnight or up to 12 hours at
36.degree. F. In some embodiments the blending times and speeds are
altered by up to 100%. In some embodiments the temperatures are
altered up to 20.degree..
[0062] Insoluble solids may be present in milks prepared by
decompounding (e.g. grinding or blending) nuts or seeds. One
surprising observation of the present invention is that these
insoluble materials can hinder the formation of curds from the
non-dairy milks. Insoluble solids can also result in curds or
cheese replicas having a texture or mouthfeel perceived as grainy
or pasty in comparison to the smoother, creamier texture of dairy
cheese. Accordingly provided herein are methods for removing these
insoluble materials and compositions of non-dairy milks and cheese
replicas with reduced amounts of the insoluble materials.
Compositions of the invention can have a mouthfeel or texture
indicative of the absence of insoluble solids. For example a cheese
replica can have a mouthfeel indicative of the absence of insoluble
solids. The insoluble materials can comprise carbohydrates.
[0063] In some embodiments the methods for producing cheese
replicas includes a step for the removal of solids from non-dairy
milks prior to curdling. For example, in some embodiments, the
non-dairy milks are centrifuged to remove the insoluble solids. In
some embodiments the present invention provides for a non-dairy
cheese replica with less than 1%, 5%, 10%, 20%, 30%, 40% or 50% of
insoluble solids found in the non-dairy milk before removal of the
insoluble solids. In some embodiments the present invention
provides for a non-dairy milk extract with less than 1%, 5%, 10%,
20%, 30%, 40% or 50% of insoluble solids found in non-dairy milk.
In some embodiments the present invention provides for a non-dairy
milk with 99%, 95%, 90%, 80%, 70%, 60% or 50% of insoluble solids
found in non-dairy milk removed.
[0064] In one embodiment the centrifugation procedure is as
follows: Pour decompounded nut or seed slurry into container. Using
a centrifuge with the JS-5.0 rotor, spin at about 5000 RPM for 30
mins. In some embodiments the spin is at about 500, 1000, 1500,
2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000,
7500, 8000, 8500, 9000, 9500, or 10,000 RPM for about 5, 10, 15,
20, 25, 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments
the centrifugation speed or time is altered, for example by
20%.
[0065] In one embodiment the centrifugation procedure is as
follows: Pour decompounded nut or seed slurry at a flow rate of
4-20 gallons per minute, for example, at a flow rate of 8 gallons
per minute, into decanter centrifuge (such as the Flottweg S4
sedicanter), spinning at a bowl speed of between 3000 and 5000 RPM.
Collect the liquid outflow for further use as a non-dairy milk, and
set aside the ejected solids.
[0066] Other methods for removal of solids from non-dairy milks
include, but are not limited to, straining, filtering, allowing to
settle, skimming, or using coagulants and flocculants (including
cations, polymer flocculants, or polyelectrolytes such as pectin,
carrageenan, alginates, or carboxymethyl cellulose) to agglomerate
particles for removal.
[0067] Centrifugation step can result in a "cream layer" and a
"skim layer". The cream layer is an emulsion comprising fats,
proteins and water. The skim layer is a solution comprising
proteins in water. In some embodiments, the cream layer and skim
layer are separated by centrifugal separation. In some embodiments,
the cream layer and skim layer are separated by centrifugal
separation in a Flotwegg ac1500 or GEA ME55. In some embodiments
the cream layer and the skim layer are incompletely separated. The
skim layer and the cream layer can be separated from the insoluble
solids in a separation process. In some embodiments the skim layer
and the cream layer are stored separately. The non-dairy milk can
comprise the skim layer. The non-dairy milk can comprise the cream
layer. Typically the skim layer and the cream layer are combined to
form the non-dairy milk. The non-dairy milk can be up to 100% cream
layer. In some embodiments the ratio of cream layer to skim layer
in the non-dairy milk is about 100:1, 90:1, 80:1, 70:1, 60:1, 50:1,
40:1, 30:1, 20:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1,
1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:20, 1:30,
1:40, 1:50, or 1:60. In some embodiments the methods described
herein comprise measuring the amount of skim layer and cream layer
being added to a non-dairy milk.
[0068] In one embodiment the separation procedure is as follows:
Remove cream layer from centrifuged bucket and place in new
container that is buried in an ice bath. Move the liquid layer
(skim layer) from the bucket into a container that is ice cold.
Keep skim and cream ice cold at all times.
[0069] In another embodiment the separation procedure is as
follows: Flow the non-dairy milk, from which insoluble solids have
been substantially removed by passage through a decanter
centrifuge, into a separator centrifuge, for example, a Flotwegg
ac1500 or GEA MESS. In some embodiments, the cream layer and skim
layer are separated by centrifugal separation in the separator
centrifuge. In some embodiments the separated skim and cream layers
are kept refrigerated.
[0070] In some embodiments the non-dairy milk is pasteurized or
sterilized. The pasteurization can be high-temperature, short-time
(HTST), "extended shelf life" (ESL) treatment, or ultra-high
temperature (UHT or ultra-heat-treated). In some embodiments the
pasteurization procedure is as follows: Pasteurize blended
non-dairy milk at 164.degree. F.-167.degree. F. for 16 seconds.
Using controlled chilling system, bring non-dairy milk temperature
down rapidly and store in a refrigerator at 36.degree. F.
[0071] Cheese replicas may also be made using non-dairy milk
prepared using proteins isolated, enriched, or purified from a
plant, or microbial source or synthesized in vitro. For example
ribulose-1,5-bisphosphate carboxylase oxygenase (rubisco) can
isolated from a plant source and included in a non-dairy milk for
the production of a cheese replica. For example 8S globulins can
isolated from a Moong beans and included in a non-dairy milk for
the production of a cheese replica. For example Pea globulins can
isolated from Pea seeds and included in a non-dairy milk for the
production of a cheese replica. For example Pea albumins can
isolated from peas and included in a non-dairy milk for the
production of a cheese replica. Isolated, enriched, or purified
plant proteins can be combined with one or more oils or fats also
isolated from plant sources, in a colloidial suspension, solution
or emulsion to form the non-dairy milk for making a cheese replica.
In some embodiments the isolated non-dairy proteins are combined
with non-non-dairy oils or fats to form a non-dairy milk. In some
embodiments multiple isolated, enriched or purified plant proteins
are used to make a non-dairy milk. Without being bound by theory
the non-dairy milks derived from isolated, enriched, or purified
proteins may reduce the problems which can be caused by insoluble
solids obtained in a slurry derived from a nut or seed. Without
being bound by theory the non-dairy milks derived from isolated,
enriched, or purified proteins may reduce the problems which can be
caused by proteins with undesirable properties, such as
lipoxygenases or proteases. Additional non-limiting examples of
proteins that may be isolated, enriched, or purified from a plant
source for the production of a non-dairy milk for cheese replica
production include: seed-storage proteins from any seed, ribosomal
proteins, actin, hexokinase, lactate dehydrogenase, fructose
bisphosphate aldolase, phosphofructokinases, triose phosphate
isomerases, phosphoglycerate kinases, phosphoglycerate mutases,
enolases, pyruvate kinases, glyceraldehyde-3-phosphate
dehydrogenases, pyruvate decarboxylases, actins, translation
elongation factors, ribulose-1,5-bisphosphate carboxylase oxygenase
(rubisco), ribulose-1,5-bisphosphate carboxylase oxygenase activase
(rubisco activase), albumins, glycinins, conglycinins, globulins,
vicilins, conalbumin, gliadin, glutelin, gluten, glutenin, hordein,
prolamin, phaseolin (protein), proteinoplast, secalin, extensins,
triticeae gluten, zein, oleosins, caloleosins, steroleosins or
other oil body proteins, vegetative storage protein A, vegetative
storage protein B, moong seed storage 8S globulin. Non-limiting
examples of oils that may be incorporated in a non-dairy milk
include: corn oil, olive oil, soy oil, peanut oil, walnut oil,
almond oil, sesame oil, cottonseed oil, rapeseed oil, canola oil,
safflower oil, sunflower oil, flax seed oil, algal oil, palm oil,
palm kernel oil, coconut oil, babassu oil, shea butter, mango
butter, cocoa butter, wheat germ oil, rice bran oil, oils produced
by bacteria, algae, archaea or fungi or genetically engineered
bacteria, algae, archaea or fungi, triglycerides, monoglycerides,
diglycerides, sphingosides, glycolipids, lecithin, lysolecithin,
phophatidic acids, lysophosphatidic acids, oleic acid, palmitoleic
acid, palmitic acid, myristic acid, lauric acid, myristoleic acid,
caproic acid, capric acid, caprylic acid, pelargonic acid,
undecanoic acid, linoleic acid, 20:1 eicosanoic acid, arachidonic
acid, eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated
linoleic acid, conjugated oleic acid, or esters of: oleic acid,
palmitoleic acid, palmitic acid, myristic acid, lauric acid,
myristoleic acid, caproic acid, capric acid, caprylic acid,
pelargonic acid, undecanoic acid, linoleic acid, 20:1 eicosanoic
acid, arachidonic acid, eicosapentanoic acid, docosohexanoic acid,
18:2 conjugated linoleic acid, or conjugated oleic acid, or
glycerol esters of oleic acid, palmitoleic acid, palmitic acid,
myristic acid, lauric acid, myristoleic acid, caproic acid, capric
acid, caprylic acid, pelargonic acid, undecanoic acid, linoleic
acid, 20:1 eicosanoic acid, arachidonic acid, eicosapentanoic acid,
docosohexanoic acid, 18:2 conjugated linoleic acid, or conjugated
oleic acid, or triglyceride derivatives of oleic acid, palmitoleic
acid, palmitic acid, myristic acid, lauric acid, myristoleic acid,
caproic acid, capric acid, caprylic acid, pelargonic acid,
undecanoic acid, linoleic acid, 20:1 eicosanoic acid, arachidonic
acid, eicosapentanoic acid, docosohexanoic acid, 18:2 conjugated
linoleic acid, or conjugated oleic acid.
[0072] In some instances the proteins used to make the non-dairy
milk may be purified proteins. Accordingly, in some embodiments the
total protein content of the cheese replica can, in some
embodiments, consist of greater than 25%, 50%, 75%, or 90%, of a
purified protein, for example any single monomeric or multimeric
protein.
[0073] Non-dairy milk compositions can also contain sugars or other
fermentable carbon sources and other nutrients. Without being bound
by theory these sugars or nutrients can favor growth of the cheese
culture microbes or serve as substrates for production of lactic
acid or other organic acids by the cheese culture microbes. For
example, in some embodiments, the non-dairy milk comprises glucose.
In other embodiments, the non-dairy milk comprises fructose. In
other embodiments, the non-dairy milk comprises sucrose. In other
embodiments, the non-dairy milk comprises high-fructose corn syrup.
In other embodiments, the non-dairy milk comprises sugar-cane
extract. In other embodiments, the non-dairy milk comprises a fruit
juice. In other embodiments, the non-dairy milk comprises
sugar-cane extract. In other embodiments, the non-dairy milk
comprises agave syrup. In other embodiments, the non-dairy milk
comprises molasses. Molasses can be cane molasses or beet molasses
or made from other non-dairy sources. In other embodiments the
non-dairy milk can comprises treacle, honey, refined sugars, or
syrup (e.g, high fructose corn syrup). In some embodiments fuel for
glycolosis, or a component (or intermediate) of the glycolysis
pathway, can be included in the compositions. Oligosaccharides can
be a part of the non-dairy milk. Disaccharides (e.g., maltose,
sucrose) can be part of a non-dairy milk. Monosaccharides (e.g.
fructose, glucose, or galactose) can be a part of the non-dairy
milk. In some embodiments the sugars are added as an addition step.
In some embodiments the sugars are not derived from the same
organism as the proteins and fats in the composition.
[0074] In some embodiments a sugar in the composition of the
invention is not treacle, honey, refined sugars, or syrup (e.g high
fructose corn syrup). In some embodiments a sugar in the
composition of the invention is not a monosaccharide (e.g.
fructose, glucose, or galactose). In some embodiments a sugar in
the composition of the invention is not a disaccharide. In some
embodiments a sugar in the composition of the invention is not an
oligosaccharide.
[0075] Non-dairy milk can, in some embodiments, contain one or more
organic acids, such as lactic acid, or acetic acid, for example to
adjust the pH and/or produce the characteristic sour taste of
cheese. These organic acids may be used in addition to or as an
alternative to microbial cultures. Accordingly, in some embodiments
the non-dairy milk comprises an organic acid. The organic acid can
be one or more of: lactic acid, acetic acid, citric acid, malonic
acid, malic acid, propionic acid.
[0076] In some instances the compositions of the invention do not
contain any animal products. In some embodiments the compositions
of the invention use no components that are derived from animals.
In some embodiments the compositions of the invention contain no
fats from animals. In some embodiments the compositions of the
invention contain no proteins from animals. In some embodiments the
compositions of the invention contain no enzymes from animals. In
some embodiments the compositions of the invention contain no dairy
products.
[0077] In one embodiment the invention provides a composition free
of or substantially free of one or more of the following: dairy
products, animal products, agar, carrageenan, or tofu.
[0078] In some embodiments the compositions have less than 20%,
less than 15%, less that 10%, less than 5%, less than 1% or less
than 0.5% starch content. In some embodiments no refined starch
(e.g. cornstarch, tapioca, wheat, or potato starch) is added to the
compositions of the invention.
[0079] In some embodiments the non-dairy milk contains no
artificial flavoring or coloring.
[0080] In some instances the non-dairy milk does contain some
animal products. For example, in some embodiments, non-dairy milk
comprises enzymes obtained from animals. In some embodiments, the
non-dairy milk contains dairy products. In some embodiments animal
fats are included in the non-dairy milk. In some embodiments the
non-dairy milk can comprise enzymes (e.g. proteases and lipases),
and/or microbes (e.g. lactic bacteria, yeast and mold). Without
being bound by theory these enzymes and or microbes can be added to
non-dairy milk formulations to produce desirable flavor and aroma
compounds.
[0081] Enzymes, including proteases and lipases, and microbes,
including lactic bacteria, yeast and mold, can be added to
non-dairy milk formulations to produce desirable flavor and aroma
compounds.
[0082] In some embodiments the invention provides a method
comprising isolating or purifying a single non-dairy protein,
mixing the isolated non-dairy protein with a non-dairy fat source,
and adding a microorganism selected from the group consisting of
Penicillium camemberti, Geotrichum candidum, Penicillium
roqueforti, Penicillium nalgiovensis, Verticillium lecanii,
Kluyveromyces lactis, Saccharomyces cerevisiae, Candida utilis,
Debaryomyces hansensil, Rhodosporidum infirmominiatum, Candida
jefer, Cornybacteria, Micrococcus sps., Lactobacillus sps.,
Lactococcus, Staphylococcus, Halomonas, Brevibacterium,
Psychrobacter, Leuconostocaceae, Streptococcus thermophilus,
Pediococcus sps., or Propionibacteria. In some embodiments at least
1 kg of the plant protein is isolated or purified from a single
non-dairy source. In some embodiments the fat is isolated from
another non-dairy source.
[0083] In some embodiments the invention provides for a composition
comprising an isolated or purified non-dairy protein and a
polynucleotide identifiable as derived from one or more of
Penicillium camemberti, Geotrichum candidum, Penicillium
roqueforti, Penicillium nalgiovensis, Verticillium lecanii,
Kluyveromyces lactis, Saccharomyces cerevisiae, Candida utilis,
Debaryomyces hansensil, Rhodosporidum infirmominiatum, Candida
jefer, Cornybacteria, Micrococcus sps., Lactobacillus sps.,
Lactococcus, Staphylococcus, Halomonas, Brevibacterium,
Psychrobacter, Leuconostocaceae, Streptococcus thermophilus,
Pediococcus sps., or Propionibacteria. In some embodiments the
cheese replicas comprise Penicillium camemberti, Geotrichum
candidum, Penicillium roqueforti, Penicillium nalgiovensis,
Verticillium lecanii, Kluyveromyces lactis, Saccharomyces
cerevisiae, Candida utilis, Debaryomyces hansensil, Rhodosporidum
infirmominiatum, Candida jefer, Cornybacteria, Micrococcus sps.,
Lactobacillus sps., Lactococcus, Staphylococcus, Halomonas,
Brevibacterium, Psychrobacter, Leuconostocaceae, Streptococcus
thermophilus, Pediococcus sps., or Propionibacteria. In some
embodiments compositions of the invention comprise Penicillium,
Geotrichum, Saccharomyces, Kluyveromyces, or Debaryomyces.
II. "Curdling" the Non-Dairy Milk
[0084] Generally the cheese-replicas are made by causing cross
linking or denaturation of proteins in non-dairy milks. These
processes replicate the curdling process of traditional dairy
cheese making.
[0085] Crosslinking is typically induced using an enzyme that
creates covalent crosslinks between polypeptide chains. In some
embodiments cross-linking enzymes are used in a cross-linking step
to curdle the non-dairy milk. In various embodiments,
transglutaminase is the cross-linking enzyme used to induce the
curdling of non-dairy milk replicas. In some embodiments the
crosslinking enzyme used to induce curdling in non-dairy milk
replicas is a lysyl oxidase. In one embodiment, the invention
provides for a method of inducing the curdling of a non-dairy milk
comprising obtaining a non-dairy milk and adding a cross-linking
enzyme to the non-dairy milk. In some embodiments the cross-linking
enzyme is transglutaminase. In some embodiments the cross-linking
enzyme is not derived from an animal source. In some embodiments
the curdling process does not use rennet.
[0086] In some embodiments between 0.1 and 20 units (U) of
transglutaminase is added per 1 mL of non-dairy milk. In some
embodiments about 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 5, 7, 10, 15, or 20
U transglutaminase is added per 1 mL of non-dairy milk. In some
embodiments the transglutaminase is followed by heated incubation,
for example in a 100.degree. F. water bath. The heated incubation
can be at a temperature optimized for the enzyme function. In some
embodiments the temperature is about 65, 70, 75, 80, 85, 90, 95,
100, 105, 110, 115, 120 or 125.degree. F.
[0087] Transglutaminases are a family of enzymes that catalyze the
formation of a covalent bond between a free amine and the
gamma-carboxyl group of glutamine thereby linking proteins
together. For example, transgluaminases catalyze crosslinking of
e.g., lysine in a protein or peptide and the gamma-carboxamide
group of a protein- or peptide-glutamine residue. The covalent
bonds formed by transglutaminase exhibit high resistance to
proteolytic degradation.
[0088] Many types of transglutaminase can be used in various
embodiments of the invention. Acceptable transglutaminases include,
but are not limited to, Streptoverticillium mobaraense
transglutaminase, an enzyme is similar or identical to a
transglutaminase from Streptoverticillium mobaraense, other
microbial transglutaminases, transglutaminases produced by
genetically engineered bacteria, fungi or algae, Factor XIII
(fibrin-stabilizing factor), Keratinocyte transglutaminase (TGM1),
Tissue transglutaminase (TGM2), Epidermal transglutaminase (TGM3),
Prostate transglutaminase (TGM4), TGM X (TGM5), TGM Y (TGM6), TGM Z
(TGM7), or lysyl oxidase. Accordingly, in some embodiments an
enzyme selected from the group consisting of a transglutaminase is
used to crosslink a non-dairy milk protein. In some embodiments the
transglutaminase is Streptoverticillium mobaraense
transglutaminase, other microbial transglutaminases,
transglutaminases produced by genetically engineered bacteria,
fungi or algae, Factor XIII (fibrin-stabilizing factor),
Keratinocyte transglutaminase (TGM1), Tissue transglutaminase
(TGM2), Epidermal transglutaminase (TGM3), Prostate
transglutaminase (TGM4), TGM X (TGM5), TGM Y (TGM6), TGM Z (TGM7),
or lysyl oxidase In some embodiments the present invention provides
for a composition comprising a non-dairy milk and
Streptoverticillium mobaraense transglutaminase, an enzyme is
similar or identical to a transglutaminase from Streptoverticillium
mobaraense, other microbial transglutaminases, transglutaminases
produced by genetically engineered bacteria, fungi or algae, Factor
XIII (fibrin-stabilizing factor), Keratinocyte transglutaminase
(TGM1), Tissue transglutaminase (TGM2), Epidermal transglutaminase
(TGM3), Prostate transglutaminase (TGM4), TGM X (TGM5), TGM Y
(TGM6) and/or TGM Z (TGM7). In some embodiments the enzyme used for
cross-linking is not Factor XIII (fibrin-stabilizing factor),
Keratinocyte transglutaminase (TGM1), Tissue transglutaminase
(TGM2), Epidermal transglutaminase (TGM3), Prostate
transglutaminase (TGM4), TGM X (TGM5), TGM Y (TGM6), TGM Z (TGM7),
or lysyl oxidase.
[0089] In some embodiments the invention provides for a composition
comprising a non-dairy milk, wherein the composition is free of
Factor XIII (fibrin-stabilizing factor), Keratinocyte
transglutaminase (TGM1), Tissue transglutaminase (TGM2), Epidermal
transglutaminase (TGM3), Prostate transglutaminase (TGM4), TGM X
(TGM5), TGM Y (TGM6) and/or TGM Z (TGM7).
[0090] Transglutaminases can be produced by Streptoverticillium
mobaraense fermentation in commercial quantities or extracted from
animal tissues. Additionally, the transglutaminase (TGM) of the
present invention may be isolated from bacteria or fungi, expressed
in bacteria or fungi from a synthetic or cloned gene, Factor XIII,
keratinocyte transglutaminase, tissue transglutaminase, epidermal
transglutaminase, prostate transglutaminase, TGM X, TGM Y, TGM Z,
or another member of the transglutaminase family. In some
particular embodiments transglutaminase is obtained from commercial
sources, for example in the form of Activa.TM. from Ajinmoto Food
Ingredients LLC.
[0091] In some embodiments the compositions of the invention have a
detectable amount of a nucleic acid from the genome of a cell that
encoded and produced an enzyme, for example a transglutaminase. For
example a cheese replica can have a detectable amount of
Streptoverticillium mobaraense DNA. This detectable amount can, for
example, be a small amount of DNA that has carried over to the
cheese replica from the production of transglutaminase by
Streptoverticillium mobaraense.
[0092] In some embodiments the cross-linking of proteins in
non-dairy milk is induced by transglutaminase. In some embodiments
the cross-linking of proteins in non-dairy milk is induced by a
lysyl oxidase. Various non-dairy milks have been crosslinked with
transglutaminase. In some embodiments the invention provides a
composition comprising a non-dairy milk and transglutaminase. In
some embodiments the invention provides a composition comprising a
curdled non-dairy milk and transglutaminase. In some embodiments
the invention provides a cheese replica containing
transglutaminase. In some embodiment the invention provides a
cheese replica with a detectable amount of degraded
transglutaminase. In some embodiment the invention provides a
cheese replica with a detectable amount of improperly folded
transglutaminase. In some embodiment the invention provides a
cheese replica with a detectable amount of transglutaminase
by-product. In some embodiment the invention provides a cheese
replica with a detectable peptide which is identifiable as portion
of a transglutaminase used in the manufacturing process for making
the cheese replica. In some embodiment the invention provides a
cheese replica with one or more detectable peptides containing a
crosslink between a lysine and glutamine residue. For example the
crosslinked glutamine and lysine residues may be detectable by mass
spectroscopy. In some embodiment the invention provides a cheese
replica with one or more detectable peptides containing a crosslink
between two lysine residues. For example the crosslinked lysine
residues may be detectable by mass spectroscopy.
[0093] Denaturation can be used instead of, or in addition to, the
cross-linking enzyme for curdling the non-dairy milk. For example,
heat denaturation of a non-dairy milk followed by cooling of the
mixture can result in a curd-like gel as, for example, described in
example 7. Accordingly, the present invention provides for
non-dairy milk curds comprising denatured non-dairy proteins. The
present invention also provides for methods for making a non-dairy
milk curd by heating a non-dairy milk to a temperature of about
between 30-35, 35-40, 40-45, 45-50, 50-55, 55-60, 60-65, 65-70,
70-75, 75-80, 80-85, 90-95, 95-100.degree. C. for about 10, 20, 30,
40, 50, 60 seconds or about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10
minutes followed by cooling. In some embodiments non-dairy milks
are curdled using both enzymatic cross-linking with
transglutaminase and heat denaturation.
[0094] Additional denaturization procedures are possible in
additional embodiments of the invention. Acids, solvents,
chaotropic agents, or disulfide bond reducers can be used to
denature the proteins in the non-dairy milk. In one embodiment urea
is added to the non-dairy milk to form curds.
III. Cheese Replica Production from the Curdled Non-Dairy Milk
[0095] Cheese replicas can be produced from the curdled non-dairy
milk. Accordingly, the invention provides methods for producing
cheese replicas from curdled non-dairy milk, as well as cheese
replica compositions.
[0096] To create the cheese replicas one might use elements of the
traditional cheese making process. Cheese replicas can be produced
using one or more steps including cutting, draining, forming,
pressing, waxing, aging, scalding, smoking, salting or
ripening.
[0097] The curd and whey may be separated in a traditional way. For
example, the cheese may be scalded to create a replica of
traditional cheddar or other hard cheese, or, the whey and curd
might be separated through a cloth drain.
[0098] The cheese replicas may be ripened in a way similar to
traditional cheese. For example, surface mould may be allowed to
grow to create a rind. In order to create a rind or color, the
process may introduce certain bacteria to the cheese replicas in
the ripening process. By way of example only, Brevibacterium linens
can be introduced to produce an orange color and pungent aroma to
the cheese replica.
[0099] In some embodiments, the cheese replica can be a salted or
salt-washed cheese replica. The salting process can preserve the
cheese replica and/or add flavor. In some embodiments the salt is
chosen based on the geographic location of where the salt is
originally obtained. In some embodiments salt is added to the cut
curds. In some embodiments salt is contacted with the outside
surface of the cheese replica, for example by rubbing. In some
embodiments the cheese replica is contacted with or soaked in
brine.
[0100] In some embodiments, the cheese replica can be a soft fresh
cheese produced by procedures exemplified by example 3.
[0101] In some embodiments, the cheese replica can be a salted or
salt-washed cheese produced by procedures exemplified by example
4.
[0102] In some embodiments, the cheese replica can be a soft
ripened or molded-rind cheese produced by procedures exemplified by
example 5.
[0103] In some embodiments, the cheese replica can be a goat cheese
replica produced by procedures exemplified by example 6.
[0104] In some embodiments, the cheese replica can have edible
materials added (e.g. herbs, pepper, spices) on its surface to
enhance flavor or add to the visual appeal of the product, as
exemplified by examples 4 and 6. In some embodiments the edible
materials are embedded in the cheese replica.
[0105] In some embodiment the cheese replica can be shaped. For
example the cheese replica can be shaped in a basket or a mold. In
some embodiments the cheese replica is pressed, for example with a
weight. The pressing can help expel any additional liquid from the
cheese replica.
[0106] In some embodiments the production of the cheese replica
includes a waxing step. In one embodiment the waxing procedure is
as follows: Cut food-grade paraffin wax into 1/2-inch pieces. Place
in double boiler and heat wax to 210.degree. F. Place cheese
replicas in standard freezer for fifteen minutes to reduce
temperature of cheese replicas to 33.degree. F. Using 3-grams of
melted wax per piece, brush wax onto cheese replicas one side at a
time. Placed waxed cheese replicas onto clean waxed paper on aging
racks. Age waxed cheese replicas in aging room at 36.degree. F.
with 75% humidity, for example for six months. In some embodiments
the aging room is between 33-70.degree. F. In some embodiments the
humidity of the aging room is altered to aid in rind formation. In
some embodiments the waxed cheese is stored for years, for example
for 2 years or more.
[0107] In some embodiments the production of the cheese replica
includes a smoking step. In some embodiments the cheese replica is
cold smoked. In some embodiments the cheese replica is smoked at
the curd stage or prior to the curd stage. In some embodiments the
cheese replica is smoked after the cheese replica is formed. In
some embodiment the smoking procedure is as follows: Soak wood
chips for six hours. Drain chips of all water and place in smoking
unit. Ignite smoker and as soon as chips have fully ignited, snuff
out flames to create smoke-filled unit. Place cheese replicas on
racks in smoker for five minutes per side. Remove from smoker and
place on cooling racks. Place cheese replicas in cooling room for
24 hours, at 36.degree. F. In various embodiments smoking times and
cooling times and temperatures will be adjusted according to the
particular cheese replica and particular desired taste profile.
[0108] In some instances the cheese replica is exposed to or
injected with mold or yeast. In some embodiments the cheese replica
is exposed to a particular bacterial strain or strains.
[0109] When the cheese replica is made without any dairy
ingredients, the production process requires no dairy farming.
Accordingly, the process for making cheese replicates is suitable
for locations where keeping animals or storing milk is difficult or
costly. In some embodiments the cheese replicas are made in
non-dairy producing regions. This will reduce the need for shipping
the products. So in some embodiments the cheese replica is a
product that is made in the same locality as its eventual
consumption. For example the cheese replica is made within 10, 20,
50, or 100 miles of its point of sale. The cheese replica
production process may also be useful in remote areas where dairy
farming is difficult or impossible. For example the cheese replica
production process could occur on an island, aboard an oil
platform, or aboard a space station.
[0110] The cheese replicas may further contain microbes useful for
cheese making. So in some embodiments the cheese replicas may be
made by a process involving culturing with any microbes used for
cheese making. The cheese replicates may be made by a process
involving fermentation. In any process, the proteins may be
crosslinked by treatment with an enzyme, which may be from the
family of transglutaminase. The cheese replicas may be created from
curds formed by a treatment of a protein-containing liquid with a
transglutaminase, or other enzymes that promote gelling, or they
may be prepared by a process involving heat denaturation of one or
more of the protein components in the solution, suspension or
emulsion to induce formation of a gel.
[0111] In some embodiments the cheese replicas comprise
Penicillium, Geotrichum, Saccharomyces, Kluyveromyces, or
Debaryomyces.
IV. Non-Dairy Cheese Replicas
[0112] The cheese replicas can be made for consumption by either
humans or other animals or both. In some instances the cheese
replicas are used to feed domesticated animals. For example, a
cheese replica of the present invention can be fed to a dog. In one
particular embodiment, the cheese replica contains a veterinary
medication In another embodiment the cheese replica comprises a
treat for a domestic animal. In some embodiments the domestic
animal treat is shaped like traditional a wedge of cheese. In some
embodiments the animal treat is artificially colored to resemble
dairy cheese, e.g. cheddar cheese.
[0113] The cheese replicas include products similar to conventional
cheese, yogurt and other dairy products. Cheese replica can be made
from a liquid extract of nuts or other fruits or seeds. The current
invention also includes cheese replicas made from proteins and fats
isolated and enriched or purified from one or more plant. The
current invention also includes cheese replicas made from proteins
and fats isolated and enriched or purified from one or more
non-dairy source.
[0114] In some embodiments the cheese replica has less than 1%, 5%,
10%, 20%, 25% or 30% polysaccharides.
[0115] In some embodiments the cheese replica has less than 1%, 5%,
10%, 20%, 25% or 30% polysaccharides because of centrifugal
separation.
[0116] In some embodiments the cheese replica has less than 1%, 5%,
10%, 20%, 25% or 30% polysaccharides because of the use of purified
proteins.
[0117] The cheese replicas may be suitable for consumption by
humans or animals who are unable to eat certain animal products,
such as a person who is lactose-intolerant or allergic to dairy.
Accordingly, in some embodiments the cheese-replica does not
contain lactose. The cheese replicas might contain enough protein
or other nutrients to be nutritionally equivalent to traditional
cheese or other animal products.
[0118] The cheese replicas may contain less fat, less saturated fat
or less cholesterol than traditional cheese or animal products, and
may be suitable for a healthier diet. So in some embodiments
provided herein is a method for distributing cheese replicas
comprising the steps of informing a lactose intolerant person of
the cheese replica, receiving an order for a cheese replica, and
making the cheese replica available to the lactose intolerant
person.
[0119] In some embodiments the lactose intolerant person is
informed through advertising. In some embodiments the lactose
intolerant person is informed by their personal physician. In some
embodiments the order for the cheese replica is sent by the lactose
intolerant person. In some embodiments the lactose intolerant
person receives the cheese replica via a distributor that places
the order. In some embodiments the informing and receiving steps
take place on a network, for instance a computer network and are
enacted with specialized computer software stored on a computer
readable media. In some embodiments the method further comprises
producing a cheese replica of the present invention and supplying
it to lactose intolerant person.
[0120] The cheese replicas are, in some embodiments, designed to
replicate the experience of eating cheese. The look, texture, and
taste of the cheese replicas can be such that it is similar or
indistinguishable from cheese. The invention provides for methods
for determining whether a animal or human can distinguish the
cheese replicas from cheese. In some embodiments a property of the
compositions of the invention is that an animal, for example a
human, will identify the composition as cheese. In some embodiments
the human identifies the composition of the invention as equivalent
to cheese. In some embodiments one or more properties of cheese are
equivalent according to an ordinary human's perception. Such
properties include the properties that can be tested listed below.
In some embodiments an ordinary human identifies a cheese replica
of the present invention as more cheese like than cheese
substitutes found in the art.
[0121] One method to determine whether the cheese replica is
comparable to cheese is to a) define the properties of cheese and
b) determine whether the cheese replicas has similar properties.
Properties of cheese that can be tested include mechanical
properties such as hardness, cohesiveness, brittleness, chewiness,
gumminess, viscosity, elasticity, and adhesiveness. Properties of
cheese or a cheese replica that can be tested also include
geometric properties such as particle size and shape, and particle
shape and orientation. Additional properties can include moisture
content and fat content. The properties of the cheese replica can
be described using terms such as "soft," "firm" or "hard" describe
hardness; "crumbly," "crunchy," "brittle," "chewy," "tender,"
"tough," "short," "mealy," "pasty," or "gummy," to describe
cohesiveness; "thin" or "viscous" or "spreadable" to describe
viscosity; "plastic" or "elastic" to describe elasticity; "sticky,"
"tacky" or "gooey" to describe adhesiveness; "gritty," "grainy" or
"coarse" or "heterogeneous" to describe particle shape and size;
"fibrous," "cellular" or "crystalline" to describe particle shape
and orientation, "dry," "moist," "wet," or "watery" to describe
moisture content; or "oily" or "greasy" to describe fat content. In
one embodiment a group of people can be asked to rate a certain
cheese, for instance cheddar, according to properties which
describe the cheese. These ratings can be used as an indication of
the properties of the cheese. The cheese replicas of the present
invention can then be compared to the properties of traditional
dairy cheese to determine how similar the cheese replica is to the
dairy cheese. In some instances the properties of the cheese
replicas are then altered to make the cheese replica more similar
to the cheese. In some embodiments, the cheese replica has a
property or properties that are rated similar to cheese according
to human evaluation. In some embodiments the cheese replica is
indistinguishable from real cheese to a human. In some embodiments
the cheese replica is distinguishable by some properties from real
cheese to a human.
[0122] In some embodiments the cheese replica is compared to real
cheese based upon olfactometer readings. In various embodiments the
olfactometer can be used to assess odor concentration and odor
thresholds, odor suprathresholds with comparison to a reference
gas, hedonic scale scores to determine the degree of appreciation,
or relative intensity of odors. In some embodiments the
olfactometer allows the training and automatic evaluation of expert
panels. In some embodiments the cheese replica is a product that
causes similar olfactometer readings to those of a particular
target cheese. In some embodiments the cheese replica is a product
that causes nearly similar, but slightly different olfactometer
readings to those of a particular target cheese.
[0123] Gas chromatography-mass spectrometry (GCMS) is a method that
combines the features of gas-liquid chromatography and mass
spectrometry to identify different substances within a test sample.
GCMS can, in some embodiments, be used to evaluate the properties
of a cheese replica. For example volatile chemicals can be isolated
from the head space around cheese. These chemicals can be
identified using GCMS. A profile of the volatile chemicals in the
headspace around cheese is thereby created. In some instances each
peak of the GCMS can be further evaluated. For instance, a human
could rate the experience of smelling the chemical responsible for
a certain peak. This information could be used to further refine
the profile. GCMS could then be used to evaluate the properties of
the cheese replicas. The GCMS could be used to refine the cheese
replica. In some embodiments the cheese replica has a GCMS profile
similar to that of cheese. In some embodiments the cheese replica
has a GCMS profile identical to that of cheese.
[0124] The cheese replicas may be modified to fit into traditional
cheese-types, including: fresh, bloomy, semi-soft, washed, firm,
hard and blue. There may be additional processes to modify the
cheese replicas and fit them into any cheese-type category. These
categories also include different presentations of the cheese
replicas, for example: sliced, shredded, block, with rind, without
rind, wet or dry. The cheese replicas may be aged, or a method may
be created to mimic the taste of traditionally-aged dairy cheese.
The cheese replicas may be classified in taste the same way
traditional cheese is. For example, they may be tart, tangy,
smooth, creamy, buttery, fluffy, rich, earthy, pungent, aromatic,
eggy, fruited, sharp, dry, caramelly, grainy, punchy or complex,
among other descriptions. The invention includes methods to modify
the cheese replicas to fit into any category or taste
description.
[0125] The cheese replicas may be modified to have or not have a
rind, may be coated in wax, and may have craters or veins typical
to blue cheese. The cheese replicas may be spreadable, such as
cream cheese. The cheese replicas can contain flavor additives, for
example: truffle, mushrooms, nuts, herbs, chives, and other
flavors.
[0126] The cheese replicas may be meltable, and have other
properties of conventional cheese.
[0127] The cheese replicas may also be similar in consistency and
taste to traditional yogurt or cottage cheese. The cheese replicas
might be flavored with fruits, sweeteners, or other flavor
additives. The cheese replicas might contain bacteria to aid with
digestion or other health additives. The cheese replicas may be
modified to have an appropriate or appealing consistency.
[0128] In various embodiments the cheese replicas may be used in
all cooking and recipes as a substitute for traditional cheese. In
some embodiments the cheese replicas may be used in cooking and
recipes as a flavor substitute for traditional cheese. In some
embodiments the cheese replicas may be used in cooking and recipes
as a functional substitute for traditional cheese.
EXAMPLES
Example 1
[0129] The Effectiveness of Transglutaminase in Curdling Various
Nut Milks was Compared.
[0130] Cashews, almonds, macadamia nuts, brazil nuts were used to
produce a slurry, by adding fresh water in a 3:1 weight ratio to
the nuts, then blending in a Vitamix 4500 blender at high speed for
2 minutes. Because we found in preliminary experiments that the
insoluble solids in the milks at this stage (slurry) inhibited
formation of optimal creamy curds and produced a cheese replica
with grainy mouth feel the slurry was subjected to centrifugation
at 10,000 G for 15 minutes at 4.degree. C. to remove insoluble
solids. The liquid supernatant, consisted of two layers, a lighter,
creamy, opaque layer and a slightly denser clear to translucent
aqueous layer. Liquid supernatant layers were recovered and blended
together and the pelleted solids were discarded. The blended
supernatants are referred to as "nut milks" (a non-dairy milk).
Each of the 4 resulting nut milks and each pair-wise combination at
a 1:1 blend, were evaluated for curdling in the presence of
transglutaminase (TG): 30 U TG was added to 15 mls of each nut milk
or blend in a 20 ml glass vial, followed by incubation in a
100.degree. F. water bath. After 5 hours all but the pure cashew
nut milk had formed sturdy curds, sufficient to retain their form
when the glass vial was inverted. The cashew milk and cashew brazil
nut milks formed soft curds. The macadamia and almond milks formed
the most solid curds. Control samples without added
transglutaminase remained liquid under these conditions.
Example 2
[0131] Preparation of Almond and Macadamia Nut Milks
[0132] Blanching (Surface Sterilization of the Nuts):
[0133] Note: Almonds and macadamia nuts were blanched
separately.
[0134] A 30 gallon tilt skillet filled to 1/2-capacity with fresh
water was heated to 212.degree. F. 25 pounds of either almonds or
macadamia nuts were dumped into the boiling water bath and left for
30 seconds. The nuts were recovered by draining into a colander
then immediately immersed in fresh ice-cold water in a second tilt
skillet to rapidly cool them. After rapid cooling the nuts were
recovered by draining in a colander and spread on sheet trays to
dry.
[0135] Macadamia nuts proceeded directly to the blending process.
Almonds were first hydrated as described below before blending.
[0136] Hydration of Almonds:
[0137] The blanched almonds were transferred to Cambro buckets and
fresh water was added to a level two inches above the level of the
nuts. The buckets were then placed at 36.degree. F. for up to 16
hours. After hydration, the nuts were recovered by draining into a
colander, rinsed with fresh water, then proceeded to the blending
step.
[0138] Blending of Nuts:
[0139] Note: Almonds and macadamia nuts were blended
separately.
[0140] One pound batches of nuts were placed in a Vitamix blender
(model Vitaprep 3), with one liter of fresh water and blended for 5
minutes, gradually increasing the blender speed to high. The
blended slurries from each batch were collected in a stainless
steel bain-marie sitting in a large container of ice and stirred
with a frozen cooling stick to cool. Once the accumulated slurry
cooled to 50.degree. F., the bain-marie was placed in a
refrigerator at 36.degree. F. for up to 12 hours.
[0141] Separation of Skim and Cream:
[0142] Sets of four 1800 ml aliquots of the blended slurry were
centrifuged at 5000 RPM/7480 G at 4.degree. C. for 30 minutes in a
Beckman-Coulter Avanti J-HC Centrifuge with a JS-5.0 rotor,
resulting in separation of the blend into 3 layers, a dense pellet
of insoluble solids, a clear to translucent aqueous layer (which we
refer to as the "skim"), and a lighter, creamy, opaque layer (which
we refer to as the "cream"). The cream layer was carefully
collected from each bucket using a spoon, and placed in a pan
sitting in a container of ice. The viscous aqueous layer (skim)
from each bucket was carefully poured into a'/2-Cambro sitting in a
container of ice. The accumulated cream and skim layers were kept
at 36.degree. F. until the pasteurization step. Typical yields were
about 0.77 lbs of cream and 0.95 lbs of skim per lb of macadamia
nuts and about 0.29 lbs of cream and 1.62 lbs of skim per lb of
almonds.
[0143] Blending
[0144] Almond skim and cream were combined in a ratio specified by
the recipe for the specific cheese replica being produced (Table
1), and blended with an immersion blender. Macadamia skim and cream
were combined in a ratio specified by the recipe for the specific
cheese replica being produced (Table 1), and blended with an
immersion blender. We hereafter refer to each blended mixture of
skim and cream as "non-dairy milk." The two non-dairy milks were
then blended in a ratio specified by the recipe for the specific
cheese replica being produced (Table 1).
[0145] Pasteurization:
[0146] The non-dairy milk blend was pasteurized at 167.degree. F.
for 16 seconds after which the non-dairy milk temperature was
rapidly reduced to 50.degree. C. in 12 seconds by a controlled
chilling system. The non-dairy milk temperature was then further
reduced by placing containers in an ice bath at 36.degree. F.
Pasteurized non-dairy milks were stored at 36.degree. F. Non-dairy
milks were discarded if the pH of the non-dairy milk dropped below
pH 6.0 during storage, indicating spoilage.
Example 3
[0147] Production of a Soft Fresh Cheese Replica.
[0148] The ingredients and amounts needed for one batch (about one
and a half pieces) of soft fresh cheese replica are listed in Table
2. The recipe can be scaled up or down proportionately.
[0149] Soft fresh cheese replica was prepared by the following
procedure:
non-dairy
[0150] Bring pasteurized non-dairy milk formula (see Table 2) to
80.degree. F. in a water bath.
[0151] Sprinkle the mesophilic starter culture (see Table 2) onto
the non-dairy milk formula and allow the culture to hydrate for
five minutes without stirring. Gently stir in the starter culture
with a spatula for two minutes. Hold at 80.degree. F. for one
hour.
[0152] Increase the water bath temperature to begin to bring the
non-dairy milk temperature to 100.degree. F.
[0153] Optional step: Add microbial "rennet" and/or distilled
vinegar (see Table 2), folding gently with a spatula. Hold for 15
minutes.
[0154] Dilute the hydrated transglutaminase (see Table 2) with a
small amount of warm non-dairy milk formula and then add it to the
non-dairy milk formula, folding gently with a spatula for two
minutes. Allow formula to reach 100.degree. F.
[0155] Remove the container of non-dairy milk formula from the
water bath and cover with plastic wrap and aluminum foil. Allow the
non-dairy milk formula to coagulate for twelve hours at room
temperature.
[0156] Cut curd into 1/2 inch dice pieces. (FIG. 1). Allow curd to
re-mat for 10 minutes. Pour the coagulated formula (curd and whey)
into a draining bag and measure its weight. Hang bag and allow curd
to drip for a minimum of 20 minutes until proper viscosity and
density of curd is achieved. The drained curd should weigh about
60% of original formula weight.
[0157] Place curd in a mixing bowl. Add cheese salt (see Table 2).
Gently whisk the curd for ten minutes, or blend for five minutes
with a Hobart mixer on low to medium speed.
[0158] Place 13.75 ounces of curd (enough to yield eight ounces
after completion of draining and brining) (FIG. 2) into a
micro-perforated mold. Drain at room temperature for one hour
without the follower in place. Then place the follower in the mold
and add 600 grams of weight to the follower. Refrigerate at
36.degree. F. for 24 hours.
[0159] Preheat saturated brine to 50.degree. F. Fully immerse the
cheese replica, still in its mold, into the preheated brine for 1/2
hour. After brining, place mold on draining rack and return it to
36.degree. F. for 24 hours.
[0160] Remove cheese replica from the mold. Place on draining mat
and return it to 36.degree. F. for 24 hours.
[0161] The soft fresh cheese replica (FIG. 3) is now ready for
packaging and shipment. Packaging process includes nitrogen flush
and airtight heat-sealing.
Example 4
[0162] Production of a Salted Cheese Replica.
[0163] The ingredients and amounts needed for one batch (about one
and a half pieces) of salted cheese replica are listed in Table 2.
The recipe can be scaled up or down proportionately.
[0164] Bring pasteurized non-dairy milk formula (see Table 2) to
80.degree. F. in a water bath.
[0165] Sprinkle the mesophilic starter culture (see Table 2) onto
the non-dairy milk formula and allow the culture to hydrate for
five minutes without stirring. Gently stir in starter culture with
a spatula for full two minutes. Hold at 80.degree. F. for one
hour.
[0166] Increase water bath temperature to begin to bring non-dairy
milk temperature to 100.degree. F.
[0167] Optional: Add microbial "rennet" and/or distilled vinegar
(see Table 2), folding gently with a spatula. Hold for 15
minutes.
[0168] Dilute the hydrated transglutaminase (see Table 2) with a
small amount of warm non-dairy milk formula and then add it to the
non-dairy milk formula, folding gently with a spatula for two
minutes. Allow formula to reach 100.degree. F.
[0169] Remove container of non-dairy milk formula from the water
bath and cover. Allow the non-dairy milk formula to coagulate for
twelve hours at room temperature.
[0170] Cut curd into 1/2 inch dice pieces (see FIG. 1). Allow curd
to re-mat for 10 minutes. Pour the coagulated formula (curd and
whey) into a draining bag and measure its weight. Hang bag and
allow curd to drip for a minimum of 20 minutes until proper
viscosity and density of curd is achieved. The drained curd should
weigh about 60% of original formula weight.
[0171] Place curd in mixing bowl. Optional: For the classic
presentation of a cheddar cheese replica (FIG. 5), annatto--a
natural vegetable dye, is added to create the yellow cheddar color.
The percentage of annatto evenly distributed throughout the curd is
0.08%. Gently whisk curd for ten minutes, or blend for five minutes
with a Hobart mixer on low to medium speed.
[0172] Place 16.3 ounces of curd (enough to yield eight ounces
after completion of draining and brining) (see FIG. 2) into a
micro-perforated mold. Drain at room temperature for one hour
without the follower in place. Then place the follower in the mold
and add 600 grams of weight to the follower. Refrigerate at
36.degree. F. for 24 hours.
[0173] Preheat saturated brine to 50.degree. F. Fully immerse the
cheese replica, still in its mold, into the preheated brine for 1/2
hour. After brining, place mold on draining rack and return it to
36.degree. F. for 24 hours.
[0174] Remove the cheese replica from the mold. Place the cheese
replica on a draining mat and return it to 36.degree. F. for 24
hours.
[0175] Move the cheese replica into a drying room for seven days at
55.degree. F., with 55% humidity, and minimal airflow. Turn the
cheese replica and place on a fresh draining mat daily.
[0176] After the seven-day drying period, remove the draining mats
and place the salted cheese replica (FIGS. 4 and 5) directly on an
aging rack.
[0177] If the cheese replica is to be waxed or smoked, return the
cheese replica to the drying room for two additional weeks at
55.degree. F., with 55% humidity and minimal airflow. Then follow
the procedure for waxing or smoking.
[0178] If the cheese replica is to be rolled in paprika and fennel
pollen mixture, follow the steps for the paprika & fennel
pollen process.
[0179] Paprika and Fennel Pollen Process:
[0180] Brush the salted cheese replica with cold water using a
pastry brush. Roll the cheese replica in 1.5 grams of a mixture of
10-parts organic paprika and 1-part fennel pollen. Place the coated
cheese replica on an aging rack and return it to the drying room
for two weeks at 55.degree. F., with 55% humidity and minimal
airflow, turning daily.
[0181] Following the drying period, move the cheese replica to
36.degree. F. for 24-hours. The paprika-fennel pollen cheese
replica (FIG. 6) is now ready for packaging and shipment.
[0182] Waxing Process:
[0183] Cut food-grade paraffin wax into 1/2-inch pieces and heat to
210.degree. F. in a double boiler.
[0184] Place the cheese replica in a freezer to reduce its
temperature to 33.degree. F. Using three grams of melted wax per
piece of cheese replica, brush wax onto cheese replica one side at
a time. Place waxed-side down onto waxed paper and then continue
brushing remainder of the cheese replica including sides and top.
Placed waxed cheese replica (FIG. 7) onto clean waxed paper on an
aging rack.
[0185] Age the waxed cheese replica in aging room at 36.degree. F.
with 75% humidity for up to six months. The waxed cheese replica is
now ready for packaging and shipment.
[0186] Smoking Process:
[0187] Soak equal parts apple and cherry wood chips for six hours.
Drain chips of all water and place in a smoking unit. Ignite smoker
and as soon as chips have fully ignited, snuff out flames to create
a smoke-filled unit. Place salted cheese on rack into smoker for
five minutes per side. Remove cheese from smoker and place on
cooling rack.
[0188] Place the cheese at 36.degree. F. for 24 hours. Smoked
cheese is now ready for vacuum aging or packaging and shipment.
Example 5
[0189] Production of a Soft Ripened Cheese Replica.
[0190] The ingredients and amounts needed for one batch (about one
and a half pieces) of soft ripened cheese replica are listed in
Table 2. The recipe can be scaled up or down proportionately.
[0191] Bring pasteurized non-dairy milk formula (see Table 2) to
90.degree. F. in a water bath.
[0192] Sprinkle lactose-free preparations of Florica Danica,
Mesophilic Starter, Geotrichum Candidum, Penicillium Candidum, and
Debaromyces hansenii (see Table 2) onto the milk formula. Optional:
Add microbial "rennet." (see Table 2). Let milk sit for 5 minutes.
Gently fold in and top stir with a spatula for two minutes. Hold at
90.degree. F. for 90 minutes.
[0193] Increase water bath temperature to start to bring non-dairy
milk temperature up to 100.degree. F.
[0194] Optional: Add in distilled vinegar (see Table 2), folding
gently with spatula. Hold 15 minutes.
[0195] Dilute the hydrated transglutaminase (see Table 2) with a
small amount of warm non-dairy milk formula and then add it to the
non-dairy milk formula, folding gently with a spatula for two
minutes. Allow formula to reach 100.degree. F.
[0196] Remove container of non-dairy milk formula from the water
bath and cover with plastic wrap and aluminum foil. Allow the
non-dairy milk formula to coagulate for twelve hours at room
temperature.
[0197] Cut curd into 1/2 inch dice pieces (see FIG. 1). Allow curd
to re-mat for 10 minutes. Pour the coagulated formula (curd and
whey) into a draining bag and measure its weight. Hang bag and
allow curd to drip for a minimum of 20 minutes until proper
viscosity and density of curd is achieved. The drained curd should
weigh about 60% of original formula weight.
[0198] Place curd in mixing bowl. Add cheese salt. Gently whisk
curd for ten minutes, or blend for five minutes with a Hobart mixer
on low to medium speed.
[0199] Place 17.64 ounces (500 grams) of curd (enough to yield
eight ounces after completion of draining and brining) (FIG. 2)
into a micro-perforated mold on a draining mat. Drain at room
temperature for one hour without the follower in place. Then place
the follower in the mold just touching the cheese replica. Do not
add any additional weight to the follower. Refrigerate at
36.degree. F. for 24 hours.
[0200] Preheat saturated brine to 50.degree. F. Fully immerse the
cheese replica, still in its mold, into the preheated brine for 1/2
hour. After brining, place mold on draining rack and return it to
36.degree. F. for 24 hours.
[0201] Remove the cheese replica from the mold. Place on draining
mat and return it to 36.degree. F. for 24 hours.
[0202] Transfer the cheese from 36.degree. F. to dry yeasting room
for three days at 60.degree. F., with 75% humidity.
[0203] After three days, transfer cheese from a draining mat to an
aging mat. Place the mat on an aging rack, and move to ripening
room at 50.degree. F., with 90% humidity and continuous airflow.
Turn the cheese replica and replace mat daily.
[0204] After seven days, transfer the cheese replica directly onto
an aging rack, allowing maximum aeration for seven more days, or
until mold coverage is complete.
[0205] After the cheese replica is thoroughly covered with mold
(FIG. 8), move it to 36.degree. F. for sixteen hours. Wrap cheese
in perforated paper and place in a wooden box for shipment.
Example 6
[0206] Production of a Goat Cheese Replica.
[0207] The ingredients and amounts needed for one batch (about one
and a half pieces) of goat cheese replica are listed in Table 2.
The recipe can be scaled up or down proportionately.
[0208] Bring pasteurized milk formula (see Table 2) to 80.degree.
F. in a water bath.
[0209] Sprinkle the mesophilic starter culture (see Table 2) onto
the non-dairy milk formula and allow the culture to hydrate for
five minutes without stirring. Gently stir in starter culture with
a spatula for full two minutes. Hold at 80.degree. F. for one
hour.
[0210] Increase water bath temperature to begin to bring non-dairy
milk temperature to 100.degree. F.
[0211] Optional: Add microbial "rennet" and/or distilled vinegar
(see Table 2), folding gently with a spatula. Hold for 15
minutes.
[0212] Dilute the hydrated transglutaminase (see Table 2) with a
small amount of warm non-dairy milk formula and then add it to the
non-dairy milk formula, folding gently with a spatula for two
minutes. Allow formula to reach 100.degree. F.
[0213] Remove container of non-dairy milk formula from the water
bath and cover with plastic wrap and aluminum foil. Allow the
non-dairy milk formula to coagulate for twelve hours at room
temperature.
[0214] Cut curd into 1/2 inch dice pieces. Allow curd to re-mat for
10 minutes. Pour the coagulated formula (curd and whey) into a
draining bag and measure its weight. Hang bag and allow curd to
drip for a maximum of 24 hours until proper viscosity and density
of curd is achieved. The drained curd should weigh about 50% of
original formula weight.
[0215] Place curd in mixing bowl. Add cheese salt (see Table 2).
Using Hobart mixer on high speed, mix and aerate completely.
[0216] Place sixteen ounces of curd (enough to yield eight ounces
after completion of draining and brining) (FIG. 2) into a
cylindrical goat cheese mold and stand the mold upright on a
draining mat. Refrigerate at 36.degree. F. for 24 hours allowing
the cheese replica to drain.
[0217] Preheat saturated brine to 50.degree. F. Fully immerse the
cheese replica, still in its mold, horizontally into the preheated
brine for 1/2 hour.
[0218] After brining, stand mold on draining mat placed at
36.degree. F. for 24 hours. Turn mold and change mat every twelve
hours for two days.
[0219] Slide shaped goat cheese replica out of mold directly onto a
bed of three grams of coarse ground pepper. Gently roll the cheese
replica in pepper, covering evenly and completely. Place the cheese
replica on waxed paper and return to 36.degree. F. for 48
hours.
[0220] The goat cheese replica (FIG. 9) is now ready for packaging
and shipment. Packaging includes Cryovac sealing process.
Example 7
[0221] Cultured Gels Made with Emulsions of Purified Plant Proteins
and Fats.
[0222] Procedure
[0223] Mixtures of purified or partially purified
protein+glucose+oil were assembled and mixed with a vortex mixer to
create an emulsion. Emulsions were either treated with
transglutaminase (Ajinomoto Activa TI at a final concentration of
2% or 4%, as noted) or first heated to 85.degree. C., rapidly
cooled on ice and then treated with transglutaminase. Mesophilic
starter culture (Danisco Choozit MA11) was added to a concentration
of 0.01% (wt/vol) concomitantly with transglutaminase to allow
cultures to ferment, acidify and impart flavors and aroma to the
gels. Reactions were incubated overnight at 30.degree. C. before
evaluating for gel formation and aroma.
[0224] Protein Purification Methods:
[0225] All steps were carried out at 4.degree. C. whenever
possible. Centrifugation steps were at 8000 g for 20 mins,
4.degree. C. Once fractionated, all ammonium sulfate precipitate
fractions of interest were stored at -20 C until further use. Prior
to their use in experiments, the precipitates were resuspended in
50 mM KPhosphate buffer pH 7.4+0.5M NaCl and dialyzed overnight
against the same buffer to remove ammonium sulfate. The dialyzed
solution was then centrifuged at 12000 g for 20 min to remove
precipitate and then used in experiments. Protein composition at
individual fractionation steps was monitored by SDS-PAGE and
protein concentrations were measured by standard UV-Vis
methods.
[0226] (i) Pea-albumins: Dry flour of green peas was used as a
source of pea albumins. The flour was resuspended in 10 volumes of
50 mM sodium acetate buffer pH 5 and stirred for 1 hr. The slurry
was centrifuged at 8000 g for 20 minutes and the supernatant was
collected. To this crude protein extract, solid ammonium sulfate
was added to 50% saturation. The solution was stirred for 1 hour
and then centrifuged. To the supernatant from this step, ammonium
sulfate was added to bring to 90% saturation. After stirring for 1
hour, the solution was stirred and then centrifuged to collect the
pea albumin proteins in the pellet. The pellet was stored at
-20.degree. C. until further use.
[0227] (ii) Pea-globulins: Dry green pea flour was used to extract
pea globulin proteins. The flour was resuspended in 10 volumes of
50 mM KPhosphate buffer pH 8+0.4M NaCl and stirred for 1 hr. After
centrifugation, the supernatant was subjected to ammonium sulfate
fractionation in two steps at 50% and 80% saturation. The 80%
pellet containing globulins of interest was stored at -20.degree.
C. until further use.
[0228] (iii) Moong bean 8S globulins: Moong bean flour was used to
extract 8S globulins by first resuspending the flour in 4 volumes
of 50 mM KPhosphate buffer pH 7 (+0.5M NaCl for lab scale
purifications). After centrifugation, proteins in the supernatant
were fractionated by addition of ammonium sulfate in 2 steps at 50%
and 90% saturation respectively. The precipitate from the 90%
fraction contained the 8S globulins and was saved at -20.degree. C.
until further use.
[0229] Gel formation was tested with the following protein
fractions:
[0230] (i) Pea-albumins (90% ammonium sulfate fraction from green
pea dry seed flour)
[0231] (ii) Pea-globulins (80% ammonium sulfate fraction from green
pea dry seed flour)
[0232] (iii) Moong bean 8S globulin (90% ammonium sulfate fraction
from moong flour)
[0233] The protein solutions were dialyzed into 50 mM KPhosphate
buffer pH 7-7.4+0.5M NaCl and used at concentrations between 8
mg/ml and 75 mg/ml (0.8-7.5%) as described below.
[0234] Canola oil was used to make emulsions. Stock solutions of
transglutaminase (Ajinomoto) were prepared at 40% in 50 mM
KPhosphate pH 7. Mesophilic starter culture (MA011 from Danisco)
was used as the cheese bacterial culture; the freeze-dried culture
was resuspended at a 10% in 50 mM KPhosphate buffer and used
immediately in the experiments. Glucose (1%) was used as sugar
source for growth of mesophilic cultures; stock solution was
prepared at 40% in water.
[0235] Two conditions were used to form gels:
[0236] (i) `Set 1`: 2% protein+3% oil+4% transglutaminase; the rest
water. Emulsify by vortex mixing.
[0237] (ii) `Set 2`: 7.5% protein+3% oil+2% transglutaminase; the
rest water. Emulsify by vortex mixing.
[0238] Both gel compositions listed above were tested for gel
formation catalyzed by transglutaminase, with and without heat
denaturation of the protein component. All reactions (total volume
of each, 1 ml) were first assembled in 1.5 ml micro-centrifuge
tubes with protein+oil+sugar components. Emulsions were made by
vortexing the samples for 30 sec. Samples to be heat denatured were
immediately placed in a water bath and heated at 85.degree. C. for
30 minutes. These samples were subsequently rapidly cooled on ice
for 20 minutes. Transglutaminase and starter culture were added to
all samples and mixed. Samples that formed gels upon heat/cool step
were vortexed to mix the enzyme and cultures in. All samples were
removed from the reaction tubes and evaluated for gel consistency
and aroma after overnight incubation at 30.degree. C.
[0239] Aroma Evaluation:
[0240] The following gels were tested for aroma (smell by 4
individuals):
[0241] Set 1: pea-albumin (heat/cool), moong (heat/cool), moong (no
heat/cool),
[0242] Set 2: pea globulin (heat/cool), moong (no heat/cool)
[0243] All four individuals reported yogurt-like/fermented/sour
aroma in the moong (set 1, no heat/cool) gels. They reported
similar smells but to a lower degree in the moong (set 2, no
heat/cool) and pea globulin (set 2, heated) gel samples. For
pea-albumins (set 1, heated) and moong (set 1, heated), very faint
or no aroma was reported. One of the four testers reported the
aroma from moong (set 2, no heat/cool)--to be unpleasant.
[0244] Gel firmness evaluation: (FIG. 10)
[0245] `Set 1`: all samples formed gels. In general these gels were
weak and disintegrated easily. Pea albumins (heat/cooled) were
slightly firmer in this regard.
[0246] `Set 2`: All samples with exception of pea-globulins (no
heat/cool) formed gels. Gels were stronger compared to Set 1 and
held their shape even after removal from the tubes. Gels formed
without heat/cool.
TABLE-US-00001 TABLE 1 MILK FORMULAS FOR SOFT FRESH, SALTED, SOFT
RIPENED AND GOAT CHEESE REPLICAS Amount Type of cheese replica milk
component (grams) Soft fresh, salted or goat Almond milk (per 800
ml batch). Almond skim 725.0 Almond Fat 75.0 Macadamia nut milk
(per 660 ml batch) Macadamia nut skim 328.5 Macadamia nut fat 337.5
Soft ripened Almond milk (per 800 ml batch). Almond skim 725.0
Almond Fat 75.0 Macadamia nut milk (per 660 ml batch) Macadamia nut
skim 291.0 Macadamia nut fat 375.0
TABLE-US-00002 TABLE 2 INGREDIENTS FOR ONE BATCH OF CHEESE REPLICA
Ingredients Amount (grams) Soft Fresh Cheese Replica: Pasteurized
almond milk 800.0 Pasteurized macadamia nut milk 666.0 Mesophilic
starter culture (MAO11 direct set) 0.5 Transglutaminase (900 units
in 15 mls of water) 15.0 Cheese salt (internal) 2.5 Salted Cheese
Replica Pasteurized almond milk 800.0 Pasteurized macadamia nut
milk 666.0 Mesophilic starter culture (MAO11 direct set) 0.5
Transglutaminase (900 units in 15 mls of water) 15.0 Soft Ripened
Cheese Replica Pasteurized almond milk 800.0 Pasteurized macadamia
nut milk 666.0 Mesophilic starter culture (MAO11 direct set) 0.3
Flora Danica 0.2 Geotrichum candidum 0.1 Penicillium candidum 0.1
Debaromyces hansenii 0.1 Transglutaminase (733 units in 15 mls of
water) 15.0 Cheese salt (internal) 4.2 Goat Cheese Replica
Pasteurized almond milk 800.0 Pasteurized macadamia nut milk 666.0
Mesophilic starter culture (MAO11 direct set) 0.5 Transglutaminase
(900 units in 15 mls of water) 15.0 Cheese salt (internal) 2.5
Optional Ingredients Distilled white vinegar 5.3 Microbial
"rennet:" 0.2 Notes: "Transglutaminase" here refers to Activa TI,
100 U/g, from Ajinomoto "Mesophilic starter culture" here refers to
Danisco Choozit lyophilized MA 11 "Geotrichum candidum" here refers
to Danisco Choozit GEO17 LYO 10D "Debaromyces hansenii" here refers
to Chr. Hansen Debaryomyces hansenii LAF3 "Flora Danica" here
refers to Chr. Hansen "Flora Danica freeze-dried lactic culture for
direct vat set" "Penicillium candidum" here refers to "Penicillium
candidum FD PC A1" from Chr. Hansen Microbial "rennet" here refers
to FROMASE 50, protease from Rhyzomicrobium mucor
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