重组β-环糊精葡萄糖基转移酶生产β-环糊精的工艺条件优化

摘要/Abstract

摘要：

将来自于Bacillus circulans 251的β-CGTase编码基因克隆到表达载体pET-20b（+），转化Escherichia coli BL21（DE3）。经酶活检测培养基上清中的β-CGTase酶活为20 U/mL。对酶转化淀粉生成β-环糊精的反应条件进行了优化，结果表明，当底物马铃薯淀粉浓度15%，反应初始pH5.5，温度30℃，加酶量10 U/g干淀粉，环己烷浓度2.5%-5%（V/V），转化周期24 h，β-环糊精转化率达到最高值75.3%，是国内外报道的酶法生产β-环糊精的最高水平。

关键词: 环糊精葡萄糖基转移酶, β-环糊精, 酶转化, 重组表达, 淀粉

Abstract:

The βcgt gene encoding β-CGTase from Bacillus circulans strain 251 was cloned into the expression vector pET-20b（+）.The vector was then transformed into Escherichia coli BL21（DE3）for extracellular production of β-CGTase. The activity in the supernatant of recombinant E. coli BL21（DE3）was 20 U/mL. Furthermore, the condition for β-cyclodextrin（β-CD）preparation by this recombinant β-CGTase was optimized. At 15% potato starch, pH5.5, 30℃, 2.5%-5%（V/V）cyclohexane, 10 U β-CGTase per gram substrate incubated for 24 hours, 75.3% of the starch was transformed into β-CD. This is the highest level of β-CD conversion by enzyme method at home and abroad.

Key words: Cyclodextrin glycosyltransferase, β-cyclodextrin, Enzymatic conversion, Recombinant expression, Starch

杨玉路, 王蕾, 陈晟, 吴敬. 重组β-环糊精葡萄糖基转移酶生产β-环糊精的工艺条件优化[J]. 生物技术通报, 2014, 0(8): 175-181.

Yang Yulu, Wang Lei, Chen Sheng, Wu Jing. Optimization of β-cyclodextrin Production by Recombinant β-cyclodextrin Glycosyltransferase[J]. Biotechnology Bulletin, 2014, 0(8): 175-181.

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