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通过双酶级联合成生物催化制备β-丙氨酸

[Biocatalytic access to β-alanine by a two-enzyme cascade synthesis].

作者信息

Gao Yu, Liu Zhongmei, Liu Ke, Zhou Zhemin, Cui Wenjing

机构信息

College of Bioengineering, Jiangnan University, Wuxi 214122, Jiangsu, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2017 May 25;33(5):875-879. doi: 10.13345/j.cjb.160416.

Abstract

Enzymatic synthesis is an important way to produce β-alanine, but the biological method is expensive generally because of the high price of substrate. In this paper, a two-step enzymatic cascade system was developed, combining L-aspartase from Escherichia coli DH5α and L-aspartate α-decarboxylase from Corynebacterium glutamicum. This system catalyzes Fumarate and ammonia to β-alanine. The optimal ratio of AspA and PanD was 1:80 (W/W), and the concentration of AspA was 10 μg/mL, at 37 ℃ and pH 7.0. When the concentration of Fumarate was 100 mmol/L, the reaction reached its equilibrium after 8 h, and the yield of β-alanine was 90 mmol/L (7 g/L). The yield of β-alanine can reach 126 mmol/L (9.8 g/L) when the concentration of Fumarate increased to 200 mmol/L. Extending reaction time, the conversion rate did not change obviously. Using this two-step enzymatic cascade system, β-alanine from cheaper substrate Fumarate can be obtained.

摘要

酶法合成是生产β-丙氨酸的重要途径,但由于底物价格高昂,生物法通常成本较高。本文构建了一种两步酶级联系统,该系统结合了来自大肠杆菌DH5α的L-天冬氨酸酶和来自谷氨酸棒杆菌的L-天冬氨酸α-脱羧酶。此系统催化富马酸和氨生成β-丙氨酸。在37℃和pH 7.0条件下,AspA和PanD的最佳比例为1:80(W/W),AspA的浓度为10μg/mL。当富马酸浓度为100mmol/L时,反应8小时后达到平衡,β-丙氨酸产量为90mmol/L(7g/L)。当富马酸浓度增至200mmol/L时,β-丙氨酸产量可达126mmol/L(9.8g/L)。延长反应时间,转化率无明显变化。利用该两步酶级联系统,可从更廉价的底物富马酸中获得β-丙氨酸。

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