Purification and characterization of a novel milk-clotting enzyme produced by Bacillus amyloliquefaciens GSBa-1

被引：25

作者：

Zhao, Xiao ^{[1
,2
]}

Cai, Miao ^{[1
]}

Yang, Zhi-Jie ^{[1
]}

Luo, Tian-Qi ^{[1
]}

Sarwar, Abid ^{[1
]}

Megrous, Sarah ^{[1
]}

Aziz, Tariq ^{[1
]}

Yang, Zhen-Nai ^{[1
,2
,3
]}

机构：

[1] Beijing Technol & Business Univ, Beijing Adv Innovat Ctr Food Nutr & Human Hlth, Beijing 100048, Peoples R China

[2] Beijing Technol & Business Univ, Beijing Engn & Technol Res Ctr Food Addit, Beijing 100048, Peoples R China

[3] Beijing Technol & Business Univ, Sch Food & Hlth, 11 Fu Cheng Rd, Beijing 100048, Peoples R China

来源：

EUROPEAN FOOD RESEARCH AND TECHNOLOGY | 2019年 / 245卷 / 11期

基金：

中国国家自然科学基金;

关键词：

Bacillus amyloliquefaciens GSBa-1; Milk-clotting enzymes; Characterization; Casein hydrolysis; Cleavage site; PROTEASE; CHEESE; RENNET; IMMOBILIZATION; COAGULANT; CLEAVAGE; SUBTILIS; CASEIN;

D O I：

10.1007/s00217-019-03361-6

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

A novel milk-clotting enzyme (MCE) produced by Bacillus amyloliquefaciens GSBa-1 was purified and identified to belong to the peptidase M4 family, and the mature peptide with milk-clotting activity (MCA) was a neutral metalloproteinase with molecular mass of about 38 kDa. The optimal pH and temperature were determined to be at 5.5 and 57 degrees C for MCA, and at 7.0 and 57 degrees C for the proteolytic activity, respectively. The MCE exhibited slight autolysis that could be inhibited by Ca2+ and Na+. Hydrolysis of caseins revealed that kappa-casein exhibited higher sensitivity to the MCE action than alpha- and beta-casein. By in-gel tryptic digestion and LC-MS/MS analysis of the major peptide (about 13 kDa) generated from hydrolysis of kappa-casein by the MCE, the cleavage site was identified to be at Lys 111-Lys 112, which was different from those of other MCEs reported earlier. The MCE from B. amyloliquefaciens GSBa-1 could serve as a novel milk coagulant for potential application in making cheese with desired proteolysis.

引用

页码：2447 / 2457

页数：11

共 38 条

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