A Fungal Alpha-Galactosidase from Pseudobalsamia microspora Capable of Degrading Raffinose Family Oligosaccharides

被引：0
作者：
Dongxue Yang
Guoting Tian
Fang Du
Yongchang Zhao
Liyan Zhao
Hexiang Wang
Tzi Bun Ng
机构：
[1] China Agricultural University,State Key Laboratory for Agrobiotechnology and Department of Microbiology
[2] Yunnan Academy of Agricultural Science,Institute of Biotechnology and Germplasmic Resource
[3] Nanjing Agricultural University,College of Food Science and Technology
[4] The Chinese University of Hong Kong,School of Biomedical Sciences, Faculty of Medicine
来源：
Applied Biochemistry and Biotechnology | 2015年 / 176卷
关键词：
Purification and characterization; Alpha-galactosidase; Chemical modification; Degradation;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
An alpha-galactosidase was purified from Pseudobalsamia microspora (PMG) to 1224.1-fold with a specific activity of 11,274.5 units/mg by ion-exchange chromatography and gel filtration. PMG is a monomeric protein with a molecular mass of 62 kDa as determined by SDS-PAGE and by gel filtration. Chemical modification using N-bromosuccinimide (NBS) resulted in a complete abrogation of the activity of PMG, suggesting that Trp is an amino acid essential to its activity. The activity was strongly inhibited by Hg2+, Cd2+, Cu2+, and Fe3+ ions. Three inner peptide sequences for PMG were obtained by liquid chromatography–tandem mass spectrometry (LC–MS–MS) analysis. When 4-nitrophenyl α-d-glucopyranoside (pNPGal) was used as substrate, the optimum pH and temperature of PMG were 5.0 and 55 °C, respectively. The Michaelis constant (Km) value of the alpha-galactosidase on pNPGal was 0.29 mM, and the maximal velocity (Vmax) was 0.97 μmol ml−1 min−1. Investigation by thin-layer chromatography (TLC) demonstrated its ability to hydrolyze raffinose and stachyose. Hence, it can be exploited in degradation of non-digestible oligosaccharides from food and feed industries.
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页码：2157 / 2169
页数：12
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