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利用顺式-环氧琥珀酸水解酶活性固定化大肠杆菌细胞生产 D(-)-酒石酸。

Immobilization of Escherichia coli cells with cis-epoxysuccinate hydrolase activity for D(-)-tartaric acid production.

机构信息

College of Life Science, Zhejiang University, Hangzhou, 310058, China.

出版信息

Biotechnol Lett. 2010 Feb;32(2):235-41. doi: 10.1007/s10529-009-0134-y. Epub 2009 Oct 21.

Abstract

Immobilization of cis-epoxysuccinate hydrolase-containing E. coli for D(-)-tartaric acid production was screened by various methods. The highest recovery of activity was obtained by entrapment in kappa-carrageenan gel. 23.6 g biomass/l and 43.4 g kappa-carrageenan/l were the best immobilization conditions optimized by response surface methodology with 83% yield (114 U/g). Cell autolysis was observed after immobilization. Immobilized cells showed high pH (5-10) stability, thermal (up to 65 degrees C) stability, conversion rate (>99.5%), enantioselectivity (ee > 99.6%), and were less affected by metal ions and surfactants compared with free cells. Conversion rate for immobilized cells preserved 93% after 10 repeated batches (5% for free cells).

摘要

采用各种方法筛选了用于 D(-)-酒石酸生产的含有顺式-环氧琥珀酸水解酶的大肠杆菌的固定化方法。通过响应面法优化,卡拉胶凝胶包埋法获得了最高的酶活回收率,最佳固定化条件为 23.6 g 生物质/L 和 43.4 g 卡拉胶/L,产率为 83%(114 U/g)。固定化后观察到细胞自溶。固定化细胞具有较高的 pH 值(5-10)稳定性、热稳定性(高达 65°C)、转化率(>99.5%)、对映体选择性(ee > 99.6%),并且与游离细胞相比,受金属离子和表面活性剂的影响较小。固定化细胞在 10 次重复批次后保留了 93%的转化率(游离细胞为 5%)。

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