U.S. patent application number 16/173414 was filed with the patent office on 2020-07-02 for tissue morphologies in filamentous fungi.
This patent application is currently assigned to Ecovative Design, LLC. The applicant listed for this patent is Ecovative Design, LLC. Invention is credited to Jacob Winiski.
Application Number | 20200208097 16/173414 |
Document ID | / |
Family ID | 55016585 |
Filed Date | 2020-07-02 |
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United States Patent
Application |
20200208097 |
Kind Code |
A1 |
Winiski; Jacob |
July 2, 2020 |
Tissue Morphologies in Filamentous Fungi
Abstract
The composite biomaterial employs a binding organism (a
filamentous fungi that produce mycelium) based on the material
physical properties required for the composite biomaterial and a
modulating organism (bacteria, fungus or yeast) based on a desired
effect of the modulating organism on the binding organism. The
modulating organism is selected based on the desired effect on the
binding organism.
Inventors: |
Winiski; Jacob; (Troy,
NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Ecovative Design, LLC |
Troy |
NY |
US |
|
|
Assignee: |
Ecovative Design, LLC
Troy
NY
|
Family ID: |
55016585 |
Appl. No.: |
16/173414 |
Filed: |
October 29, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
14745881 |
Jun 22, 2015 |
10125347 |
|
|
16173414 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C12N 1/18 20130101; C12N
1/14 20130101; C12N 1/20 20130101 |
International
Class: |
C12N 1/14 20060101
C12N001/14; C12N 1/20 20060101 C12N001/20; C12N 1/18 20060101
C12N001/18 |
Claims
1. A composite biomaterial, wherein the biomaterial comprises:
discrete particles; a binding organism, wherein the binding
organism binds the discrete particles; and a modulating organisms,
wherein the modulating organism is selected to induce aerial growth
in the binding organism.
2. The composite biomaterial of claim 1, wherein the modulating
organism is homogeneously dispersed with the binding organism in
the composite biomaterial.
3. The composite biomaterial of claim 1, wherein the modulating
organism and the binding organism have one or more interfaces in
the composite biomaterial.
4. The composite biomaterial of claim 3, wherein the binding
organism exhibits aerial growth in response to the modulating
organism at the one or more interfaces.
5. The composite biomaterial of claim 1, wherein the modulating
organism further induces the binding organism to produce thickened
mycelium.
6. The composite biomaterial of claim 1, wherein the binding
organism is selected from the group consisting of: Ganoderma
tsugae, Trametes hirsuta and Ganoderma Oregonense.
7. The composite biomaterial of claim 1, wherein the modulating
organism is selected from the group consisting of: Rhizopus sp. and
Mucor sp., Aureobasidium sp., Trichoderma sp., Penecillium sp.,
Chlysonillia sp., and Aspergillus sp., and Phanerochaete sp.,
Trichaptum sp., Stereum sp., Phlebia sp., Laetiporus sp., and
Peniophora sp.
8. The composite biomaterial of claim 1, wherein the discrete
particles comprise lignocellulose.
9. A composite biomaterial of claim 1 wherein the binding organism
produces pigment.
10. The composite biomaterial of claim 9, wherein the modulating
organism is homogeneously dispersed in the composite
biomaterial.
11. The composite biomaterial of claim 9, wherein the modulating
organism and the binding organism have one or more interfaces in
the composite biomaterial.
12. The composite biomaterial of claim 11, wherein the binding
organism produces pigment in response to the modulating organism at
the one or more interfaces.
13. The composite biomaterial of claim 9, wherein the modulating
organism produces pigment in response to the binding organism.
14. The composite biomaterial of claim 9, wherein the binding
organism is selected from the group consisting of: Ganoderma
tsugae, Trametes hirsuta and Ganoderma Oregonense.
15. The composite biomaterial of claim 9, wherein the modulating
organism is selected from the group consisting of: Rhizopus sp. and
Mucor sp., Aureobasidium sp., Trichoderma sp., Penecillium sp.,
Chlysonillia sp., and Aspergillus sp., and Phanerochaete sp.,
Trichaptum sp., Stereum sp., Phlebia sp., Laetiporus sp., and
Peniophora sp.
16. The composite biomaterial of claim 9, wherein the discrete
particles comprise lignocellulose.
Description
[0001] This invention claims priority of Provisional Patent
Application 62/021,240 filed Jul. 7, 2014 and s Division of U.S.
Ser. No. 14/745,881, filed Jun. 22, 2015
[0002] This invention relates to a composite biomaterial composed
of discrete particles, a binding organism and a modulating
organism.
BACKGROUND OF THE INVENTION
[0003] In nature, fungi produce a number of biochemical and
morphological responses to other microorganisms as a means of
competing for resources. These interactions typically fall into two
categories: antagonism from a distance (in which antagonistic
bioactive compounds are leached into the environment to inhibit the
growth of surrounding organisms), and physical antagonism (in which
the fungus comes into physical antagonistic contact with a
competing microorganism).
[0004] Fungi also have a dynamic range of responses to the
antagonism of competing microorganisms, including rapid recovery,
fruiting and sporulation, and the creation of physical barriers
between the mycelial mass and the competing organism (such as
thickened zones of mycelium at the interface, extra-cellular
pigmentation/melanin).
[0005] As is known, U.S. Pat. No. 9,485,917 describes various
techniques for making a biomaterial composed of a substrate of
discrete particles and a network of interconnected mycelia cells
extending through and around the discrete particles and bonding
discrete particles together. This biomaterial leverages the
tenacious strength of fungal vegetative mycelium.
[0006] It is an object of the invention to obtain specific tissue
morphologies in the fungus used for making composite materials by
placing the binding organism in competitive contact with the
modulating organism.
[0007] It is another object of the invention to selectively inhibit
growth of the binding organism via application of the modulating
organism to the binding organism.
[0008] Briefly, the invention provides a composite biomaterial made
in accordance with a method for stimulating the expression of
specific tissue morphologies in filamentous fungi via interactions
with competing microorganisms. These tissue morphologies are
described within the context of biomaterial production utilizing
the vegetative mycelium of filamentous fungi, and the morphologies
produced via the described competitive interactions may provide
unique material physical properties within the said context.
Relationships and interactions are described between A) a binding
organism (the filamentous fungus being cultivated as a
biomaterial), and B) a modulating organism (a microorganism
introduced to the binding organism in order to elicit the
expression of tissue morphology specific to the competitive
interaction of the binding organism with the modulating organism
and/or to completely inhibit the growth of the binding organism as
a means of selectively controlling the boundaries of growth of the
binding organism.
[0009] The process stimulates multiple tissue morphologies, with
each providing particular functionalities within a given
mycelium-based biomaterial based on the organisms selected and the
method of modulating organism dispersal in relation to the binding
organism. The specific interactions, and the results of those
specific interactions, are additionally controlled by the selection
of organisms based on A) application, B) environmental context, C)
nutritive context, and D) ecological context.
[0010] In particular, the invention provides a composite
biomaterial made by a method comprising the steps of selecting a
binding organism based on the material physical properties required
for the composite biomaterial and selecting a modulating organism
based on a desired effect of the modulating organism on the binding
organism.
[0011] The method requires the steps of inoculating a mass of
discrete substrate particles with the binding organism; applying
the modulating organism to the combination of binding organism and
discrete substrate particles; and incubating the combination of
binding organism, discrete substrate particles and modulating
organism in an environment that is conducive to the growth of both
the binding organism and modulating organism to produce a composite
biomaterial having the discrete substrate particles bound together
by the binding organism and the modulating organism imparting said
desired effect on the binding organism.
[0012] These and other objects of the invention will become more
apparent from the following detailed description taken in
conjunction with the accompanying drawings wherein:
[0013] FIG. 1 illustrates a schematic representation of a
cross-section of a binding organism on a modulating organism with a
growth boundary therebetween;
[0014] FIG. 2 illustrates a schematic representation of a binding
organism growing without the presence of a modulating organism;
[0015] FIG. 3 illustrates a schematic representation of a binding
organism growing with the presence of a modulating organism;
[0016] FIG. 4 illustrates a schematic representation of a binding
organism expanding over a modulating organism on a plate; and
[0017] FIG. 5 illustrates a schematic representation of primordium
development in response to a modulating organism;
[0018] Referring to FIG. 1, by placing a binding organism 10 on a
modulating organism 11 in binary opposition to one another a clear
boundary of interaction is created at the interface. This can
function as a method of describing specific growth boundaries for
the binding organism; the binding organism can be restricted to
growing and expanding within defined parameters within a given
volume or two-dimensional plane.
[0019] Referring to FIG. 2, wherein like reference characters
indicate like parts as above, a binding organism 10 growing on a
plate 12 without a modulating organism provides a smooth-looking
appearance at a top surface.
[0020] Referring to FIG. 3, wherein like reference characters
indicate like parts as above, the binding organism 10 is grown with
the modulating organism 11 homogeneously disbursed throughout the
growth medium. In this case, the binding organism 10 is the only
growth visually apparent, but the overall density of growth has
been reduced due to the interaction of the modulating organism
11.
[0021] The "growth medium" is the nutrient substrate that the
binding and modulating organisms are growing on. This is most often
discrete lignocellulose particles.]
[0022] By homogeneously disbursing the modulating organism 11
throughout the volume of the growing binding organism 10, the
overall density of the binding organism's mycelial colonization can
be reduced via the competitive action of the modulating organism
11.
[0023] This method can be used for producing mycelium-based
materials that require a reduced density of mycelial colonization
without the need for chemical, nutritive, or environmental
retardation of colonization.
[0024] Referring to FIG. 4, wherein like reference characters
indicate like parts as above, by controlling the capacity of the
modulating organism 11 to utilize the nutrition and environmental
context, while also controlling the selection of binding organism
10 and modulating organism 11, a relationship can be developed that
results in the binding organism 10 expressing a denser quality of
mycelial colonization when in contact with the modulating organism
11 than would be typical. This can be utilized for increasing the
density of growth for a given mycelium-based biomaterial, thereby
modulating the material's flexural strength, compressive strength,
thermal characteristics or stiffness.
[0025] As illustrated, the binding organism 10 has expanded over
top of the modulating organism 11 (located at 4 points along the
outside of the culture plate 12), leading to denser mycelial
colonization of the binding organism 10 when coming into contact
with the modulating organism 11.
[0026] When placed in binary opposition either the binding organism
10, the modulating organism 11, or both organisms can produce
extra-cellular pigments 13 (such as melanin] at the interface
between the two organisms. Based on species selection, this
interaction can produce specific aesthetic results, which can be
used to pigment the surface (or selected areas of the surface] of a
given mycelium-based biomaterial.
[0027] A pigmentation response during binary interaction of a
binding organism 10 with a modulating organism 11 resulted in a
very dense production and depositing of melanin 13 at an interface
between the two organisms.
[0028] Referring to FIG. 5, wherein like reference characters
indicate like parts as above, by either placing the binding
organism 10 and modulating organism 11 in a binary interaction, or
by otherwise disbursing the modulating organism 11 throughout the
mass of the binding organism 10, the induction of
primordia/sporocarps can occur. Depending on the maturity and
location of sporocarps on the given mycelium-based biomaterial, the
strength and/or cushioning characteristics of the material can be
increased.
[0029] As illustrated, in response to the modulating organism 11,
the binding organism 10 (the only growth visually apparent)
expanded and covered a selectively placed modulating organism 11,
at which point, the binding organism 10 developed a large
primordium 14 in the exact footprint of the modulating organism
12.
[0030] Based on organism selection, and placement of the binding
organism 10 with the modulating organism 11, a thickened and/or
aerial quality of vegetative growth can be induced at the interface
between the binding organism 10 and the modulating organism 11. In
this case, the thickened mycelium can impart additional cushioning
or strength characteristics to the given mycelium-based
biomaterial.
[0031] By homogeneously disbursing the modulating organism 11
throughout the binding organism 10, or selectively disbursing the
modulating organism 11, and depending on organism selection, a
generalized aerial quality of vegetative growth can be achieved.
This aerial growth can provide additional cushioning and aesthetic
characteristics to a given mycelium-based biomaterial.
[0032] A "fuzzy" aerial mycelium expressed across the surface of a
mycelium-based biomaterial via the homogeneous disbursing of a
modulating organism 11 throughout the volume of the binding
organism 10.
[0033] The method for stimulating the expression of specific tissue
morphologies in filamentous fungi comprises the following process
steps: [0034] 1. Select a binding organism based on the desired
material performance, as well as nutritive and environmental
context for cultivation. Examples of a binding organism would be
filamentous fungi that produce mycelium that has attractive
material physical properties based on the intended application (for
example, high tensile strength for application as a low-density
packaging material). Particularly, a filamentous fungi from the
Basidiomycetes, such as Ganoderma tsugae, Trametes hirsuta, or
Ganoderma oregonense. [0035] 2. Select a modulating organism based
on the desired effect on the binding organism, desired method of
disbursal, and nutritive and environmental context for cultivation.
The modulating organisms ecological niche in relation to the
binding organism's ecological niche should be considered, as well
as the modulating organisms ability to utilize the nutritive and
environmental context. Examples of a modulating organism would be
fungi or bacteria that specifically demonstrate a
competitive/antagonistic dynamic with the binding organism.
Examples include, but are not limited to: [0036] Bacteria:
Pseudomonas sp., and Bacillus sp. [0037] Fungi: Zygomycetes, such
as Rhizopus sp. and Mucor sp., Ascomycetes, such as Aureobasidium
sp., Trichoderma sp., Penecillium sp., Chrysonillia sp., and
Aspergillus sp. Basidiomycetes, such as Phanerochaete sp.,
Trichaptum sp., Stereum sp., Phlebia sp., Laetiporus sp., and
Peniophora sp. Yeast, such as Saccharomyces sp. [0038] 3. Inoculate
the target media for colonization/supporting mycelia colonization
with the binding organism. This is the discrete particles to be
bound together into a composite material by the mycelium of the
binding organism, most often discrete lignocellulose particles
(such as agricultural by-products or wood particles). [0039] 4.
Disburse the modulating organism in relation to the binding
organism by: [0040] a.) Placing the two organisms in a binary
interaction, creating a single interface. [0041] b.) Homogeneously
disbursing the modulating organism throughout the mass of the
binding organism. [0042] c.) Selectively placing the modulating
organism through, on, or in opposition to a portion of the overall
mass of the binding organism via 4a or 4b. [0043] 5. Incubate the
combination of binding and modulating organism within the desired
environmental conditions until the desired quality of mycelial
growth and tissue morpohology has been achieved. This step is
highly specific to the intended application. For instance, if the
intention is to induce primordia along the interface between the
modulating and binding organism to provide a cushioning
characteristic, and primordia 2 cm tall are required to achieve the
desired cushioning characteristic, incubation would continue until
the primordia reach a height of 2 cm. [0044] 6. Further process the
biomaterial as necessary This is the composite material made up of
the discrete lignocellulose particles bound together by the
mycelium of the binding organism (which has had a particular
expression induced by the modulating organism, and/or had its
growth boundaries defined by inhibition of the modulating
organism).
EXAMPLES
Example 1
Selection of Binding and Modulating Organism
[0045] 1] Select nutrition for supporting the growth of the binding
organism
[0046] 2] Select a binding organism appropriate for the nutrition
and environment for cultivation. For example, if using virgin
lignocellulose a primary wood decomposing species should be
selected.
[0047] 3a] Select a modulating organism with an inferior ability to
exploit the nutrition selected as compared to the binding organism.
For example, if using pine substrate a species that cannot utilize
pine, or utilize pine to the extent as the selected binding
organism, should be selected.
[0048] 3b] Select a modulating organism with an inferior growth
rate than that of the selected binding organism. In this instance
the binding organism out-paces the modulating organism, leading to
domination of resources/nutrition.
[0049] 3c] Select a modulating organism that is significantly
effected by the competitive/antagonistic behavior of the binding
organism. In this base the binding organism out-competes the
modulating organism, leading to domination of
resources/nutrition.
[0050] 3d] Select a modulating organism that is inappropriate for
the environmental conditions for colonization. For example, if the
temperature for cultivation is 80 F, and the binding organism grows
appropriately at 80 F, then a modulating organism that does not
grow appropriately at 80 F should be selected.
Example 2
Disbursal of a Modulating Organism through, or in Relation to, the
Mass of the Binding Organism
[0051] 1] Inoculate a given mass of lignocellulose substrate with a
primary saprophyte binding organism.
[0052] 2a] Homogeneously mix the modulating organism inoculum with
the lignocellulose substrate.
[0053] 2b] Homogeneously mix the modulating organism inoculum into
a selected portion of the lignocellulose substrate.
[0054] 2c] Apply the modulating organism inoculum to the entire
surface, or a selected portion of the entire surface of the
lignocellulose substrate mass.
[0055] 3] Incubate the inoculated lignocellulose until the desired
quality of colonization and tissue morphology has been
achieved.
[0056] 4] Further process the mycelium biomaterial as desired.
Example 3
Using Binary Stand-Off to Define Growth Boundaries for the Binding
Organism within a Mass of Lignocellulose Substrate
[0057] 1] In layers apply lignocellulose substrate to a bed.
[0058] 2] Selectively disburse the binding organism inoculum into
the positive space (area in 2-dimensions where the binding organism
is desired to grow].
[0059] 3] Selectively disburse the modulating organism inoculum
into the negative space (area in 2-dimensions where the binding
organism is not desired to grow). A modulating organism that cannot
utilize the lignocellulose nutrition being utilized should be
selected.
[0060] 4] Repeat steps 1-3 until a volume of lignocellulose
substrate has been built, with the volume of desired binding
organism growth defined in three dimensions within the mass via the
selective placement of binding and modulating organism
inoculum.
[0061] 5] Incubate the lignocellulose mass until the binding
organism has reached the desired stage of colonization and
expresses the desired tissue morphology.
[0062] 6] Remove the colonized substrate from the non-colonized
substrate.
[0063] 7] Further process the colonized mass as desired.
Example 4
Decreasing or Increasing the Density of Mycelial Colonization
within a Biomaterial Consisting of Mycelium Binding Together
Discreet Particles of Lignocellulose
[0064] 1] Select a binding organism appropriate for the target
substrate for colonization.
[0065] 2a] To increase density select a modulating organism
previously determined to induce an increase in mycelial density in
the binding organism.
[0066] 2b] To decrease the density select a modulating organism
previously determined to induce a decrease in mycelial density in
the binding organism.
[0067] 3] Incubate the inoculated substrate until the desired
quality of colonization has been achieved.
[0068] 4] Process the colonized substrate as desired.
Example 5
Inducing Pigmentation on the Surface of a Mycelium-Based
Biomaterial
[0069] 1] Select a binding organism that is both appropriate for
the target substrate intended for colonization and has been
previously identified as producing pigmentation in the vegetative
mycelium.
[0070] 2] Follow the steps of modulating organism distribution and
incubation of any of examples 1-4.
Example 6
Inducing Sporocarp Development on the Surface of a Mycelium-Based
Biomaterial
[0071] 1] Select a binding organism that is both appropriate for
the target substrate and has been shown to develop primordia in
response to competition with a modulating organism.
[0072] 2] Follow the steps of modulating organism distribution and
incubation of any of examples 1-5.
Example 7
Inducing Thickened and/or Aerial Mycelium on the Surface/through
the Volume of a Biomaterial Consisting of Mycelium Binding together
Discreet Particles of Lignocellulose
[0073] 1] Select a binding organism that is both appropriate for
the target substrate and has been shown to develop aerial
vegetative mycelium in response to competition with a modulating
organism.
[0074] 2] Follow the steps of example 2.
* * * * *