US 11,951,155 B2
Pea-derived peptide with muscle-building effect and preparation method thereof, and drug and use
Zhao Zhang, Heze (CN); Jiuxun Zhang, Heze (CN); Zheng Zhang, Heze (CN); Jie Li, Heze (CN); Wei Wei, Heze (CN); Xiping Zhang, Heze (CN); Zhongli Pei, Heze (CN); Shuai Gu, Heze (CN); and Lizhu Niu, Heze (CN)
Assigned to Zhongshi Duqing(Shandong) Biotech Co., Ltd, Heze (CN)
Filed by Zhongshi Duqing(Shandong) Biotech Co., Ltd, Heze (CN)
Filed on Aug. 22, 2022, as Appl. No. 17/821,421.
Claims priority of application No. 202210248928.6 (CN), filed on Mar. 15, 2022.
Prior Publication US 2023/0293631 A1, Sep. 21, 2023
Int. Cl. A61K 38/16 (2006.01); A61P 21/06 (2006.01)
CPC A61K 38/168 (2013.01) [A61P 21/06 (2018.01)] 15 Claims
 
1. A method for preparing a pea-derived peptide comprising one or more of the following polypeptides as separate peptide fragments: PP1 with an amino acid sequence shown in SEQ ID NO: 1, PP2 with an amino acid sequence shown in SEQ ID NO: 2, PP3 with an amino acid sequence shown in SEQ ID NO: 3, PP4 with an amino acid sequence shown in SEQ ID NO: 4, and PP5 with an amino acid sequence shown in SEQ ID NO: 5, wherein the method comprises the following steps:
1) conducting enzymatic hydrolysis on a pea protein under an action of an alkaline protease to obtain a first enzymatic hydrolysate;
2) conducting enzymatic hydrolysis on the first enzymatic hydrolysate under an action of papain and cellulase to obtain a second enzymatic hydrolysate;
3) inactivating the second enzymatic hydrolysate, then filtering inactivated second enzymatic hydrolysate through a ceramic membrane, filtering through an organic membrane, and collecting a permeate to obtain a pea-derived peptide;
4) subjecting the pea-derived peptide to gel chromatography, and collecting a fraction S2 12-18 min;
5) subjecting the fraction S2 to separation and purification by preparative liquid chromatography, and collecting a fraction W5 at 55-63 min; and
6) subjecting the fraction W5 to separation and purification by analytical liquid chromatography, and sequentially collecting 5 polypeptide fragments at a PP1 time interval, a PP2 time interval, a PP3 time interval, a PP4 time interval, and a PP5 time interval, respectively, to obtain PP1, PP2, PP3, PP4, and PP5.