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食品与发酵工业  2017, Vol. 43 Issue (6): 54-    
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调控NADH/NAD+对重组谷氨酸棒杆菌产L-丝氨酸的影响
朱加粉,陈紫薇,张晓梅,史劲松,许正宏
江南大学药学院,江苏无锡,214122
江南大学生物工程学院,江苏无锡,214122
The effect of NADH/NAD+ regulation on L-serine production by recombinant Corynebacterium glutamicum
ZHU Jia-fen,CHEN Zi-wei,ZHANG Xiao-mei,SHI Jin-song,XU Zheng-hong
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摘要 在谷氨酸棒杆菌中,L-丝氨酸由糖酵解中间产物3-磷酸甘油酸经过3步反应生成,这个过程产生2个NADH,而L-丝氨酸的合成过程并不涉及NAD+的生成,多余的NADH是否会影响菌株的生长及产L-丝氨酸?该研究通过外源添加NAD+的前体物质烟酸,内源表达NADH氧化酶编码基因noxA,考察调控NADH/NAD+对Corynbacterium glutamicum SYPS-062 33a△SSAAI生长和产L-丝氨酸的影响.结果表明,添加不同浓度烟酸,菌株的L-丝氨酸产量、生物量及糖耗较未添加时略有提高.而通过加强表达NADH氧化酶编码基因noxA,构建重组菌Corynbacterium glutamicum SYPS-062 33a△SSAAI-noxA,重组菌中NADH氧化酶比酶活是出发菌的11.6倍,L-丝氨酸产量达到28.93 g/L,较出发菌提高9.0%,糖酸转化率达到0.29 g/g蔗糖,较出发菌提高7.4%,0D562最大值为51.38,较出发菌提高6.6%,说明NADH氧化酶的过表达能够促进重组菌株的生长及碳源利用,提高菌株的L-丝氨酸产量.
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Abstract: The L-serine is produced by a three-step reaction of a glycolytic intermediate 3-phosphoglycerate,2 molecules of NADH were generated in the production of L-serine from sugars by Corynebacterium glutamicum.However,no NAD + was generated in this process.Did NADH affect L-serine production and cell growth of Corynebacterium glutamicum This study aimed to investigate the effects of regulation of NADH/NAD + on Corynebacterium glutamicum SYPS-062 33a△SSAAI growth,sugar consumption and L-serine accumulation by exogenous addition of niacin and endogenous overexpression of NADH oxidase.The results showed that exogenous addition of niacin had slightly enhanced the cell growth,sugar consumption and L-serine accumulation.When NADH oxidase was overexpressed,the recombinant strain Corynebacterium glutamicum SYPS-062 33a△SSAAI-noxA was constructed.The NADH oxidase activity in the recombinant strain was 11.6 folds higher than that of the parent strain.The production of L-serine in recombinant strain was 28.93 g/L,increased by 9.0%,and the yield was 0.29 g/g sugar,increased by 7.4%.Its OD562 was 51.38,increased by 6.6%.The results indicated that the overexpression of NADH oxidase could increase the cell growth,sugar consumption and L-serine production.
               出版日期:  2017-06-25      发布日期:  2017-12-01      期的出版日期:  2017-06-25
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