吐温-80对钝齿棒杆菌产L-赖氨酸的影响

doi:10.7506/spkx1002-6630-201623029

摘要/Abstract

摘要： 对吐温-80提高L-赖氨酸产量的分子机制进行探究。以钝齿棒杆菌MT-M4 ΔproB为出发菌株，在菌株生长至对数初期36 h时，添加5 mg/mL吐温-80进行摇瓶发酵，通过反转录实时荧光定量聚合酶链式反应（reversetranslate real-time fluorescence quantitative polymerase chain reaction，RT-qPCR）监控相关基因的转录水平、胞内的还原型烟酰胺腺嘌呤二核苷酸磷酸（nicotinamide adenine dinucleotide phosphate reduced form，NADPH）水平及α-酮戊二酸脱氢酶复合物（α-oxoglutarate dehydrogenase complex，ODHC）的活力。摇瓶发酵结果表明，添加吐温-80使L-赖氨酸产量提高了215%；RT-qPCR结果表明，编码ODHC的基因odhA、sucB和lpdA、参与ODHC调控的基因pknG、dtsR和ppp以及合成NADPH的磷酸戊糖途径基因zwf的转录水平均发生了变化，其中odhA、sucB、lpdA、pknG及dtsR1基因分别发生了上调，ppp基因下调，相应ODHC活力提高了32.9%；zwf基因上调了17.6 倍，相应胞内NADPH水平提高了137%。因此，在对数生长初期36 h添加吐温-80具有双重功效，既提高了ODHC的表达又使NADPH积累，从而诱导L-赖氨酸的高产。

关键词: 钝齿棒杆菌, 吐温-80, L-赖氨酸, &alpha, -酮戊二酸脱氢酶复合物, NADPH

Abstract: The current work aimed to investigate the molecular mechanisms of Tween-80 for triggering L-lysine
overproduction by Corynebacterium crenatum. L-lysine yield was increased by approximately 215% with the addition
of Tween-80 in C. crenatum. By using reverse translate real-time fluorescence quantitative polymerase chain reaction,
we determined that the transcriptional levels of odhA, sucB and lpdA, which encode three subunits of 2-oxoglutarate
dehydrogenase complex (ODHC), were up-regulated; pknG and dtsR1 involved in the signal transduction pathway for
ODHC regulation were up-regulated but ppp was down-regulated. As a result, ODHC-specific activity was increased by
32.9%. Furthermore, the transcriptional level of zwf involved in the pentose phosphate pathway was increased by 17.6
folds; thus, intracellular NADPH level was enhanced by 137% in C. crenatum triggered by Tween-80. The addition of
Tween-80 to the fermentation medium at the early exponential growth phase induced a bifunctional mechanism for L-lysine
overproduction by improving ODHC activity and NADPH supply in C. crenatum.

Key words: Corynebacterium crenatum, Tween-80, L-lysine, 2-oxoglutarate dehydrogenase complex, NADPH

中图分类号:

Q939.9

赖木兰，万方，朱蔷云，陈雪岚. 吐温-80对钝齿棒杆菌产L-赖氨酸的影响[J]. 食品科学, doi: 10.7506/spkx1002-6630-201623029.

LAI Mulan, WAN Fang, ZHU Qiangyun, CHEN Xuelan. Effect and Molecular Mechanisms of Tween-80 on L-lysine Production in Corynebacterium crenatum[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201623029.

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吐温-80对钝齿棒杆菌产L-赖氨酸的影响

Effect and Molecular Mechanisms of Tween-80 on L-lysine Production in Corynebacterium crenatum

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