# US 9,751,929 B2 · Method of using papain to extract collagen from aquatic animals

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- Number: US 9,751,929 B2
- Document ID: US9751929B2
- Status: Granted
- Kind: B2
- Application number: 14/170,396
- Filed: 2014-01-31
- Published: 2014-05-29
- Granted: 2017-09-05
- Earliest claimed: 2011-06-13
- Assignees: Universiti Putra Malaysia Serdang
- Applicants: Universiti Putra Malaysia Serdang
- Inventors: Jamilah Bakar; Umi H. Razali; Dzulkifly M. Hashim; Awis Q. Sazili; Kaur Harvinder
- Web page: https://patentgrep.com/patent/US9751929B2

## CPC

- C07K 14/78: Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 14/461: {from fish}
- G01N 33/6887: {from muscle, cartilage or connective tissue}
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

Disclosed is a method of using papain to extract collagen from the skin of aquatic animals. The method comprising using papain to obtain collagen by introducing the collagen fibers into a sodium chloride solution; collecting the collagen fibers and centrifuging the collagen fibers for at least 20 minutes; and obtaining a collagen yield of between 14 and 40% by weight.

## Claims

### Claim 1

A process of extracting collagen from skins of fish selected from the group consisting of Lates calcarifer (barramundi) and Oreochromis nilotica (red tilapia) using papain, comprising:
a) homogenizing the fish skin with sodium hydroxide;
b) obtaining a suspension from step (a);
c) stirring the suspension for at least 24 hours;
d) centrifuging the suspension from step (c) for at least 20 minutes;
e) separating a precipitate from step (d);
f) homogenizing the precipitate from step (e) with alkaline solution;
g) stirring the precipitate from step (f) for at least 24 hours and for at least 3 repeats;
h) washing the precipitate with water and acetic acid solution;
i) stirring the precipitate from step (h) with papain at a working temperature of 5° C. for at least 24 hours;
j) separating a papain-treated precipitate from step (i);
k) centrifuging the papain-treated precipitate from step (j) for at least 20 minutes;
l) separating collagen fibers from the centrifuged papain-treated precipitate of step (k);
m) introducing the collagen fibers into a sodium chloride solution;
n) collecting the collagen fibers from step (m) and centrifuging the collagen fibers for at least 20 minutes;
o) separating the resulting collagen pellet;
p) freeze-drying the collagen fibers from step o; wherein the collagen yield of said skins is between 14 and 40% by weight.

### Claim 2 (depends on 1)

The process of claim 1, wherein said skin is obtained by cutting the skin from fresh or frozen fish.