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Position: Home > Articles > Studies on the Gene Cloning and its Characterization of α-Galactosidases from Brachybacterium sp. DB5 Current Biotechnology 2015 (04) 310-314

一种来源于Brachybacterium sp.DB5的α-半乳糖苷酶克隆及性质研究

作  者:
刘小丹;董继胜;姬铁强;杨培龙
单  位:
中国农业科学院饲料研究所
关键词:
α-半乳糖苷酶;短状杆菌;基因克隆;表达;性质
摘  要:
α-半乳糖苷酶是一种有着巨大商业价值的工业酶制剂,在医药、食品和化工等行业有着广泛的应用。以来源于雪莲根部土壤的短状杆菌Brachybacterium sp.DB5为材料,从其基因组中扩增出一个α-半乳糖苷酶基因编码序列,经过测序及BLAST比对分析,证实该基因属于α-半乳糖苷酶。将其与p ET-30a(+)载体相连后在大肠杆菌中进行异源表达,经过诱导获得了此酶的胞外高效表达,粗酶液的活性为5.07 U/m L,经纯化后酶的活性达72.78 U/m L,酶学特性分析表明其最适p H为6.0,最适温度为40℃。此酶可用作动物饲料豆粕的添加剂,以提高饲料的利用率。
译  名:
Studies on the Gene Cloning and its Characterization of α-Galactosidases from Brachybacterium sp. DB5
作  者:
LIU Xiao-dan;DONG Ji-sheng;JI Tie-qiang;YANG Pei-long;Heilongjiang Dilong Pharmaceutical Co. Ltd;Feed Research Institute,Chinese Academy of Agricultural Siences;
关键词:
α-galactosidase;;Brachybacterium sp.;;gene cloning;;expression;;characterization
摘  要:
Alpha- galactosidase is a kind of industrial enzyme preparation with a huge commercial value,which has been widely used in medicine,food and chemical industries. A strain of Brachybacterium sp. DB5 was selected to isolate α- galactosidase gene by PCR. Sequencing and BLAST analysises showed that the sequence encoded a α-galactosidase. It was recombinant into the vector p ET-30a( +) and transformed into the strain E. coli. α-Galactosidase gene was expressed efficiently after induction with IPTG. An expression activity 5. 07 U / m L was obtained. After purification of pure,enzyme activity was 72. 78 U / m L. The enzymatic analysis revealed that its optimal p H and temperature were 6.0 and 40℃,respectively. The enzyme could be used for additive of animal feed,soybean meal,in order to improve the feed utilization rate.

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