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.

引用

页码：98 / 104

页数：7

共 47 条

[21] Biotechnological potential of microbial α-galactosidases [J].

Katrolia, Priti ;

Rajashekhara, Eranna ;

Yan, Qiaojuan ;

Jiang, Zhengqiang .

CRITICAL REVIEWS IN BIOTECHNOLOGY, 2014, 34 (04) :307-317

[22] Characterization of a protease-resistant α-galactosidase from the thermophilic fungus Rhizomucor miehei and its application in removal of raffinose family oligosaccharides [J].

Katrolia, Priti ;

Jia, Huiyong ;

Yan, Qiaojuan ;

Song, Shuang ;

Jiang, Zhengqiang ;

Xu, Haibo .

BIORESOURCE TECHNOLOGY, 2012, 110 :578-586

[23] PURIFICATION AND PROPERTIES OF ALPHA-GALACTOSIDASE FROM ESCHERICHIA-COLI SUBSP COMMUNIOR IAM-1272 [J].

KAWAMURA, S ;

KASAI, T ;

TANUSI, S .

AGRICULTURAL AND BIOLOGICAL CHEMISTRY, 1976, 40 (04) :641-648

[24]

King MR, 1998, FEMS MICROBIOL LETT, V163, P37, DOI 10.1016/S0378-1097(98)00148-7

[25]

Kumar S, 2016, MOL BIOL EVOL, V33, P1870, DOI [10.1093/molbev/msw054, 10.1093/molbev/msv279]

[26] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

[27] IMMOBILIZED ALPHA-D-GALACTOSIDASE IN THE SUGAR-BEET INDUSTRY [J].

LINDEN, JC .

ENZYME AND MICROBIAL TECHNOLOGY, 1982, 4 (03) :130-136

[28] Gene cloning, expression and characterization of an α-galactosidase from Pedobacter nyackensis MJ11 CGMCC 2503 with potential as an aquatic feed additive [J].

Liu, Xiaodan ;

Meng, Kun ;

Wang, Yaru ;

Shi, Pengjun ;

Yuan, Tiezheng ;

Yang, Peilong ;

Luo, Huiying ;

Bai, Yingguo ;

Yao, Bin .

WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY, 2009, 25 (09) :1633-1642

[29]

LOWRY OH, 1951, J BIOL CHEM, V193, P265

[30] Molecular cloning and characterization of a novel α-galactosidase gene from Penicillium sp F63 CGMCC 1669 and expression in Pichia pastoris [J].

Mi, Shijun ;

Meng, Kun ;

Wang, Yaru ;

Bai, Yingguo ;

Yuan, Tiezheng ;

Luo, Huiying ;

Yao, Bin .

ENZYME AND MICROBIAL TECHNOLOGY, 2007, 40 (05) :1373-1380

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