Characterization of a novel GH36 α-galactosidase from Bacillus megaterium and its application in degradation of raffinose family oligosaccharides

被引：44
作者：
Huang, Yan [1 ]
Zhang, Han [1 ]
Ben, Peipei [1 ]
Duan, Yajuan [1 ]
Lu, Meiling [2 ]
Li, Zhoukun [1 ]
Cui, Zhongli [1 ]
机构：
[1] Nanjing Agr Univ, Coll Life Sci, Key Lab Agr Environm Microbiol, Minist Agr, Nanjing, Jiangsu, Peoples R China
[2] China Pharmaceut Univ, Sch Life Sci & Technol, Nanjing, Jiangsu, Peoples R China
来源：
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES | 2018年 / 108卷
关键词：
alpha-Galactosidases; Bacillus megaterium; Protease resistance; High hydrolytic activity; Raffinose family oligosaccharides; FLATULENCE-CAUSING FACTORS; TRANSGLYCOSYLATION ACTIVITY; MOLECULAR-CLONING; HETEROLOGOUS EXPRESSION; RHIZOMUCOR-MIEHEI; PICHIA-PASTORIS; PROTEASE; PURIFICATION; REMOVAL; SOYMILK;
D O I：
10.1016/j.ijbiomac.2017.11.154
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
A novel alpha-galactosidase gene (agaB) from Bacillus megaterium 3-7 was cloned and expressed in Escherichia coli. The gene coded for a protein with 741 amino acids and a calculated molecular mass of 85.4 kDa. The native structure of the recombined AgaB was determined to be a homotrimer. AgaB showed the highest identity of 57% with the characterized glycosyl hydrolase family 36 alpha-galactosidase from Clostridium stercoraritan F-9. The enzyme exhibited a specific activity of 362.6 U/mg at 37 degrees C and pH 6.8. The enzyme showed strong resistance to proteases and great tolerance to galactose (K-i = 12.5 mM). AgaB displayed wide substrate specificity toward pNPGal, melibiose, raffinose and stachyose, with a K-m of 0.42, 12.1, 17.0 and 25.4 mM, respectively. Furthermore, AgaB completely hydrolyzed raffinose and stachyose present in soybean milk at 37 degrees C within 4 h when combined with trypsin. These favorable properties make AgaB a potential candidate for applications in the food and feed industries. (C) 2017 Elsevier B.V. All rights reserved.
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收藏
页码：98 / 104
页数：7
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