# US 10,501,509 B2 · Method of preparing functional peptides from germinated beans

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- Number: US 10,501,509 B2
- Document ID: US10501509B2
- Status: Granted
- Kind: B2
- Application number: 15/404,154
- Filed: 2017-01-11
- Published: 2017-10-12
- Granted: 2019-12-10
- Earliest claimed: 2016-04-11
- Assignees: Zhejiang Academy of Agricultural Sciences
- Applicants: Zhejiang Academy of Agricultural Sciences
- Inventors: Wei Wang; Nan Wang; Yu Zhang; Junhong Wang; Zuoyi Zhu; Xue Li; Jiahong Zhu
- Web page: https://patentgrep.com/patent/US10501509B2

## CPC

- C07K 14/415: from plants
- C12P 21/06: produced by the hydrolysis of a peptide bond, e.g. hydrolysate products (preparing foodstuffs by protein hydrolysis A23J3/00)
- 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)
- 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)}

## Abstract

The present invention discloses a method of preparing a functional peptide from a germinated bean, including the steps of: germinating a bean until a sprout length is about 0.5-1.0 cm; adding water to the obtained germinated bean, and then blending and crushing the germinated bean; adjusting the obtained slurry to a pH value between 6.5-8.0, and then adding neutral protease, papain and compound flavor protease to perform enzymatic reaction under stirring condition, wherein an enzymatic reaction solution is obtained including the functional peptide; subjecting the enzymatic reaction solution to ultrafiltration using an ultrafiltration membrane; keeping the ultrafiltrated enzymatic reaction solution at a temperature between 70-80° C. or using ozonated water at a concentration between 20-25 mg/L thereby avoiding decomposition by microorganisms; and performing spray drying to obtain the functional peptide.

## Claims

### Claim 1

A method of preparing a functional peptide from a germinated bean, comprising the following steps:
(1) germinating a bean to obtain a germinated bean with a sprout length of 0.5-1.0 cm;
(2) adding water to the germinated bean obtained from step (1), and then blending and crushing the germinated bean to obtain a slurry, wherein the germinated bean and the water are in a weight ratio of 1:0.9-1.1;
(3) adjusting the slurry obtained from step (2) to a pH value between 6.5-8.0, and then adding neutral protease, papain and compound flavor protease to incubate under stirring condition at a temperature between 50-55° C. for 2.5-3.5 hours to obtain an enzymatic reaction solution comprising the functional peptide;
wherein the neutral protease and the germinated bean obtained from step (1) are in a weight ratio of 1:10, the papain and the germinated bean obtained from step (1) are in a weight ratio of 1:10, and the compound flavor protease and the germinated bean obtained from step (1) are in a weight ratio of 1:10;
(4) heating the enzymatic reaction solution obtained from step (3) to a temperature between 85-98° C. and keeping the enzymatic reaction solution at the temperature for 10-15 minutes, and then cooling the heated enzymatic reaction solution to a room temperature to terminate the enzymatic reaction, and then centrifuging the cooled enzymatic reaction solution at 2500-3500 r/min for 15-25 minutes, wherein a supernatant of the centrifugation contains the functional peptide;
(5) subjecting the supernatant obtained from step (4) to ultrafiltration using an ultrafiltration membrane at an operating pressure between 0.1-0.2 MPa and an operating temperature between 20-45° C. to obtain an ultrafiltrated solution, wherein the ultrafiltration membrane has a molecular weight cut-off of 5 kDa;
(6) keeping the ultrafiltrated solution at a temperature between 70-80° C.; and
(7) performing spray drying to obtain the functional peptide.

### Claim 2 (depends on 1)

The method according to claim 1, wherein inlet air of the spray drying is controlled at a temperature between 180-190° C. and outlet air of the spraying drying is controlled at a temperature between 95-110° C.

### Claim 3 (depends on 1)

The method of claim 1, wherein the neutral protease has an enzyme activity of 500,000 u/kg, the papain has an enzyme activity of 3,500,000 u/g and the compound flavor protease has an enzyme activity of 1000 LAPU/g.

### Claim 4 (depends on 1)

The method of claim 1, wherein the bean is soybean.