Zhang Jiandong, Cui Zhimei, Chang Honghong, Fan Xiaojun, Zhao Qiuyong, Wei Wenlong
Department of Biological and Pharmaceutical Engineering, College of Chemistry and Chemical Engineering, Taiyuan University of Technology, No.79 West Yingze Street, Taiyuan, 030024, Shanxi, People's Republic of China.
Biotechnol Lett. 2016 Sep;38(9):1559-64. doi: 10.1007/s10529-016-2130-3. Epub 2016 May 27.
To investigate the efficiency of a cofactor regeneration enzyme co-expressed with a glycerol dehydrogenase for the production of 1,3-dihydroxyacetone (DHA).
In vitro biotransformation of glycerol was achieved with the cell-free extracts containing recombinant GlyDH (glycerol dehydrogenase from Escherichia coli), LDH (lactate dehydrogenase form Bacillus subtilis) or LpNox1 (NADH oxidase from Lactobacillus pentosus), giving DHA at 1.3 g l(-1) (GlyDH/LDH) and 2.2 g l(-1) (GlyDH/LpNox1) with total turnover number (TTN) of NAD(+) recycling of 6039 and 11100, respectively. Whole cells of E. coli (GlyDH-LpNox1) co-expressing both GlyDH and LpNox1 were constructed and converted 10 g glycerol l(-1) to DHA at 0.2-0.5 g l(-1) in the presence of zero to 2 mM exogenous NAD(+). The cell free extract of E. coli (GlyDH-LpNox) converted glycerol (2-50 g l(-1)) to DHA from 0.5 to 4.0 g l(-1) (8-25 % conversion) without exogenous NAD(+).
The disadvantage of the expensive consumption of NAD(+) for the production of DHA has been overcome.
研究与甘油脱氢酶共表达的辅因子再生酶用于生产1,3 - 二羟基丙酮（DHA）的效率。
含有重组甘油脱氢酶（来自大肠杆菌的甘油脱氢酶）、乳酸脱氢酶（来自枯草芽孢杆菌的乳酸脱氢酶）或LpNox1（来自戊糖乳杆菌的NADH氧化酶）的无细胞提取物实现了甘油的体外生物转化，分别以1.3 g l⁻¹（甘油脱氢酶/乳酸脱氢酶）和2.2 g l⁻¹（甘油脱氢酶/LpNox1）的产量产生DHA，NAD⁺循环的总周转数（TTN）分别为6039和11100。构建了共表达甘油脱氢酶和LpNox1的大肠杆菌全细胞（GlyDH - LpNox1），在存在0至2 mM外源NAD⁺的情况下，将10 g甘油l⁻¹转化为0.2 - 0.5 g l⁻¹的DHA。大肠杆菌（GlyDH - LpNox）的无细胞提取物在无外源NAD⁺的情况下，将甘油（2 - 50 g l⁻¹）转化为0.5至4.0 g l⁻¹的DHA（转化率为8 - 25%）。
克服了生产DHA时NAD⁺消耗昂贵的缺点。
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