Characterization of an acidic alpha-galactosidase from hemp (Cannabis sativa L.) seeds and its application in removal of raffinose family oligosaccharides (RFOs)

文献类型： 外文期刊

第一作者： Zhang, Weiwei

作者： Zhang, Weiwei;Du, Fang;Zhang, Weiwei;Du, Fang;Zhang, Weiwei;Wang, Hexiang;Zhang, Weiwei;Wang, Hexiang;Tian, Guoting;Zhao, Yongchang;Ng, Tzi Bun

作者机构：

关键词： alpha-galactosidases; Cannabis sativa L.; purification; raffinose family oligosaccharides

期刊名称：ACTA BIOCHIMICA POLONICA （ 影响因子：2.149； 五年影响因子：2.175 ）

ISSN： 0001-527X

年卷期： 2018 年 65 卷 3 期

页码：

收录情况： SCI

摘要： An acidic alpha-galactosidase designated as hemp seed alpha-galactosidase (HSG) was purified from hemp (Cannabis sativa L.) seeds. By means of chromatographic procedures which involved chromatography on the cation-exchangers CM-cellulose and SP-Sepharose, chromatography on the anion-exchangers DEAE-cellulose and Q-Sepharose, and gel filtration on Superdex 75 using fast protein liquid chromatography, HSG was purified to electrophoretic homogeneity. Results of SDS-PAGE and gel filtration on FPLC Superdex 75 revealed that the enzyme was a monomeric protein with a molecular weight of 38 kDa. Sequences of the inner peptides of the alpha-galactosidase obtained by MALDI-TOF-MS showed that HSG was a novel alpha-galactosidase since there was a little similarity to the majority of alpha-galactosidases recorded in the literature. A pH of 3.0 and a temperature of 50 degrees C were optimal for the activity of the enzyme. The activity of HSG was inhibited by the chemical modification with N-bromosuccinimide (NBS) reagent. HSG contained 16 tryptophan residues and two tryptophan residues on the surface, which were crucial to the alpha-galactosidase activity. The heavy metal ions Cd2+, Cu2+, Hg2+ and Zn2+ inhibited its activity. The K-m and V-max for the hydrolysis of pNPGal (4-nitrophenyl alpha-D-galactopyranoside) were respectively 0.008 mM and 68 mu M min(-1) mg(-1). HSG also catalyzed the hydrolysis of raffinose and other natural substrates. Hence the alpha-galactosidase possesses a tremendous potential for food and feed industries in the elimination of indigestible oligosaccharides from leguminous products.

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