Production of L-malic acid with fixation of HCO3- by malic enzyme-catalyzed reaction based on regeneration of coenzyme on electrode modified by layer-by-layer self-assembly method
Production of L-malic acid with fixation of HCO3- by malic enzyme-catalyzed reaction based on regeneration of coenzyme on electrode modified by layer-by-layer self-assembly method
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DOI：
10.1016/j.jbiosc.2008.09.009
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发表时间：
2009-01-01
影响因子：
2.8
通讯作者：
Suye, Shin-ichiro
中科院分区：
文献类型：
--
作者：
Zheng, Haitao;Ohno, Yoko;Suye, Shin-ichiro
从缺陷短波单胞菌（Brevundimonas diminuta）IFO 13182中分离纯化的苹果酸酶，以NAD（+）为辅酶，催化苹果酸脱羧生成丙酮酸和CO2，并以HCO 3-为碳源，利用其逆反应合成L-苹果酸。L-苹果酸的生产是基于NADH在碳板电极上的电化学再生，所述碳板电极通过逐层吸附聚合物结合的介体（藻酸结合的紫精衍生物，Alg-V）、聚合物结合的辅酶（藻酸结合的NAD（+），Alg-NAD（+））和硫辛酰胺脱氢酶（LipDH）来修饰。在多层膜中实现了LipDH催化固定化NAD（+）的电化学还原，采用Alg-V/LipDH/Alg-NAD（+）/苹果酸酶多层膜在电极上逐层固定的HCO 3固定体系生产L-苹果酸，L-苹果酸产量接近11.9 mmol，HCO 3固定率接近47.4%在仅含有KHCO 3和异辛酸钾盐的缓冲液中，使用阳离子交换膜。在48 h内，NADH的总转换数约为19，000，表明在多层膜内实现了有效的NADH再生和快速的电子转移，并且该修饰电极是一种潜在的固定HCO 3-而不添加游离辅酶的方法。(C)2008年，日本生物技术学会。All rights reserved.
Malic enzyme prepared and purified from Brevundimonas diminuta IFO13182 catalyzed the decarboxylation reaction of malate to pyruvate and CO2 using NAD(+) as the coenzyme, and the reverse reaction was used in the present study for L-malic acid production with fixation of HCO3- as a model compound for carbon source. The L-malic acid production was based on electrochemical regeneration of NADH on a carbon plate electrode modified by layer-by-layer adsorption of polymer-bound mediator (Alginic acid bound viologen derivative, Alg-V), polymer-bound coenzyme (Alginic acid bound NAD(+), Alg-NAD(+)), and lipoamide dehydrogenase (LipDH). Electrochemical reduction of immobilized NAD(+) catalyzed by LipDH in a multilayer film was achieved, and the L-malic acid production with HCO3- fixation system with layer-by-layer immobilization of Alg-V/LipDH/Alg-NAD(+)/malic enzyme multilayer film on the electrode gave an L-malic acid production of nearly 11.9 mmol and an HCO3- fixation rate of nearly 47.4% in a buffer containing only KHCO3 and pyruvic acid potassium salt, using a cation exchange membrane. The total turnover number of NADH within 48 h was about 19,000, which suggests that efficient NADH regeneration and fast electron transfer were achieved within the multilayer film, and that the modified electrode is a potential method for the fixation of HCO3- without addition of free coenzyme. (C) 2008, The Society for Biotechnology, Japan. All rights reserved.