# US 2021/0340587 A1 · Process for production of keratin microfibers and protein hydrolysate from poultry feathers via microbial hydrolysis

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- Number: US 2021/0340587 A1
- Document ID: US20210340587A1
- Status: Published application
- Kind: A1
- Application number: 17/283,575
- Filed: 2019-10-17
- Published: 2021-11-04
- Earliest claimed: 2018-10-17
- Assignees: Bioextrax AB
- Applicants: Bioextrax AB
- Inventors: Mohammad H.A. Ibrahim
- Web page: https://patentgrep.com/patent/US20210340587A1

## CPC

- C12P 21/06: produced by the hydrolysis of a peptide bond, e.g. hydrolysate products (preparing foodstuffs by protein hydrolysis A23J3/00)
- C07K 14/78: Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
- C08H 1/06: derived from horn, hoofs, hair, skin or leather
- C08L 89/04: Products derived from waste materials, e.g. horn, hoof or hair
- C12N 1/20: Bacteria; Culture media therefor
- C12P 13/04: Alpha- or beta- amino acids {(other amino acids C12P13/005)}
- C12R 2001/07: Bacillus
- C12R 2001/085: Bacillus cereus
- C12R 2001/10: Bacillus licheniformis
- C12R 2001/11: Bacillus megaterium
- C12R 2001/125: Bacillus subtilis {; Hay bacillus; Grass bacillus}
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C08H: DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS (polysaccharides C08B; natural rubber C08C; natural resins or their derivatives C09F; bituminous materials C10)
- C08L: COMPOSITIONS OF MACROMOLECULAR COMPOUNDS (compositions based on polymerisable monomers C08F, C08G; artificial filaments or fibres D01F; textile treating compositions D06)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The present invention relates to a process of converting feather materials into keratin-rich microfibers and soluble protein hydrolysate, the process comprising using a microbial hydrolysis technique.

## Claims

### Claim 1

A process for production of keratin microfibers from feather material, the process comprising:
a) Providing a fermentation medium;
b) Providing feather material at a concentration of at least 25 grams dry weight/L of fermentation medium;
c) Providing a keratin-degrading bacteria;
d) Contacting the fermentation medium, the feather material and the keratin-degrading bacteria, and thereby fermenting the feather material to obtain a fermented composition;
e) Separating the fermented composition obtained in step d) into one supernatant fraction and one precipitate fraction, wherein the precipitate fraction obtained in the separation step comprises keratin microfibers,
thereby obtaining keratin microfibers, wherein said keratin microfibers comprise at least 75% dry weight keratin, and wherein said microfibers have a length of between 20 μm and 200 μm.

### Claim 2 (depends on 1)

The process according to claim 1, wherein the feather material is provided at a concentration of 40 g dry weight/L or above.

### Claim 3 (depends on 1, 2)

The process according to claim 1 or claim 2, wherein the feather material is the sole source of carbon and/or nitrogen for the keratin-degrading bacteria.

### Claim 4

The process according to any one of the preceding claims, wherein the precipitate fraction obtained in the separation step further comprises fermenting cells.

### Claim 5

The process according to any one of the preceding claims, wherein the separation step comprises centrifugation and/or filtration.

### Claim 6

The process according to any one of the preceding claims, wherein the process further comprises a step of purifying the precipitate fraction.

### Claim 7

The process according to any one of the preceding claims, wherein the step of purifying comprises:
re-suspending the precipitate fraction, and/or washing the precipitate fraction and/or mixing the precipitate fraction with water;
centrifugation and/or filtration of the re-suspended, washed and/or mixed precipitate fraction.

### Claim 8

The process according to any one of the preceding claims, wherein the purification step results in one supernatant fraction and one precipitate fraction, and wherein the precipitate fraction obtained in the purification step comprises purified keratin microfibers, and the supernatant fraction obtained in the purification step comprises fermenting cells.

### Claim 9

The process according to any one of the preceding claims, wherein the purification step is optionally repeated on the precipitate fraction at least twice, such as at least three times.

### Claim 10

The process according to any one of the preceding claims, wherein the process further comprises a step of drying the precipitate fraction obtained from the separation step and/or from the purification step.

### Claim 11

The process according to any one of the preceding claims, wherein the supernatant fraction obtained in the purification step comprises fermenting cells and is recirculated to the fermenting step.

### Claim 12

The process according to any one of the preceding claims, wherein the feather material comprises or consists of feathers.

### Claim 13

The process according to any one of the preceding claims, wherein the feather material comprises quill and/or vane of feathers.

### Claim 14

The process according to any one of the preceding claims, wherein the feathers are selected from the group consisting of vaned feathers, down feathers, pennaceous feathers, contour feathers, filoplume, remiges and flight feathers.

### Claim 15

The process according to any one of the preceding claims, wherein the feathers are from poultry, such as from chicken and/or turkey.

### Claim 16

The process according to any one of the preceding claims, wherein said keratin-degrading bacteria belongs to the genus Bacillus.

### Claim 17

The process according to any one of the preceding claims, wherein the keratin-degrading bacteria is selected from the group consisting of Bacillus pumilus, B. subtilis, B. megaterium, B. licheniformis, and B. cereus.

### Claim 18

The process according to any one of the preceding claims, wherein the keratin-degrading bacteria is Bacillus pumilus.

### Claim 19

The process according to any one of the preceding claims, wherein the keratin-degrading bacteria is Bacillus pumilus FH9, deposited with the Deutsche Sammlung von Microorganismen and Zellkulturen GmbH on 24 Mar. 2014 under deposition number DSM 28594.

### Claim 20

The process according to any one of the preceding claims, wherein the fermenting step is aerobic.

### Claim 21

The process according to any one of the preceding claims, wherein the temperature of the fermenting step is between 30° C. and 42° C., such as between 32° C. and 38° C., such as between 34° C. and 38° C., such as between 36° C. and 38° C., such as 37° C.

### Claim 22

The process according to any one of the preceding claims, wherein pH of the fermenting step is between pH 5 to pH 8, such as between pH 6 to pH 9, such as between pH 6.5 to pH 8.5, such as pH 8.

### Claim 23

The process according to any one of the preceding claims, wherein the time for fermentation is 12 h to 96 h, such as 12 h to 84 h, such as 24 h to 84 h, such as 36 h to 84 h, such as 48 h to 72 h, such as 24 h.

### Claim 24

The process according to any one of the preceding claims, wherein the fermenting step comprises mechanical stirring.

### Claim 25

A microfiber composition comprising keratin microfibers, wherein said keratin microfibers comprise at least 75% dry weight keratin, and wherein said keratin microfibers have a length of between 20 μm and 200 μm and a diameter of between 1 μm and 10 μm, and wherein said keratin microfibers comprise hollow cylindrical structures.

### Claim 26 (depends on 23)

The keratin microfiber composition according to claim 23, wherein said microfibers comprise between 75% dry weight keratin and 100% dry weight keratin, such as between 80% dry weight keratin and 100% dry weight keratin, such as between 85% dry weight keratin and 100% dry weight keratin, such as between 75% dry weight keratin and 95% dry weight keratin, such as between 80% dry weight keratin and 95% dry weight keratin, such as between 85% dry weight keratin and 95% dry weight keratin.

### Claim 27 (depends on 23, 24)

The keratin microfiber composition according to any one of claims 23 and 24, wherein said microfibers have a length of between 25 μm and 200 μm, such as of between 30 μm and 200 μm, such as of between 35 μm and 200 μm, such as of between 40 μm and 200 μm, such as of between 50 μm and 200 μm, such as of between 60 μm and 200 μm, such as of between 70 μm and 200 μm, such as of between 80 μm and 200 μm, such as of between 90 μm and 200 μm, such as of between 100 μm and 200 μm, such as of between 20 μm and 180 μm, such as of between 20 μm and 150 μm, such as of between 20 μm and 120 μm, such as of between 20 μm and 100 μm, such as of between 20 μm and 90 μm, such as of between 20 μm and 80 μm, such as of between 20 μm and 70 μm, such as of between 20 μm and 60 μm, such as of between 30 μm and 150 μm.

### Claim 28 (depends on 23, 24, 25)

The keratin microfiber composition according to any one of claims 23 to 25, wherein said keratin microfibers have a diameter of between 1 μm and 6 μm, such as of between 2 μm and 6 μm, such as of between 3 μm and 6 μm, such as of between 4 μm and 6 μm, such as of between 5 μm and 6 μm, such as of between 1 μm and 5 μm, such as of between 1 μm and 4 μm, such as of between 1 μm and 3 μm, such as of between 1 μm and 2 μm.

### Claim 29 (depends on 23, 24, 25, 26)

The keratin microfiber composition according to any one of claims 23 to 26, wherein said keratin microfibers are insoluble in an aqueous solution.

### Claim 30 (depends on 23, 24, 25, 26, 27)

The keratin microfiber composition according to any one of claims 23 to 27, wherein each keratin-rich microfiber comprises at least one internode and at the most seven internodes, such as at least one internode and at the most six internodes, such as at least one internode and at the most five internodes.

### Claim 31 (depends on 23, 24, 25, 26, 27, 28)

The keratin microfiber composition according to any one of claims 23 to 28, wherein said keratin microfibers comprise between 75% dry weight proteins and 100% dry weight proteins, such as between 80% dry weight proteins and 100% dry weight proteins, such as between 85% dry weight proteins and 100% dry weight proteins, such as between 75% dry weight proteins and 90% dry weight proteins, and wherein said keratin-rich microfibers comprise between 0.1% dry weight lipids and 2% dry weight lipids, such as about 1% dry weight lipids.

### Claim 32 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 23, 24, 25, 26, 27, 28, 29)

The keratin microfiber composition according to any one of claims 23 to 29, wherein said microfiber composition is obtained by the process of any one of claims 1 to 21.

### Claim 33 (depends on 23, 24, 25, 26, 27, 28, 29, 30)

Use of the keratin microfiber composition according to any one of claims 23 to 30, for manufacture of a composite material.

### Claim 34 (depends on 31)

The use according to claim 31, wherein the keratin microfiber composition is used for manufacture of a homocomposite material, such as a keratin homocomposite material.

### Claim 35 (depends on 31)

The use according to claim 31, wherein the keratin microfiber composition is used for manufacture of a microfiber-reinforced plastic.