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通过常压室温等离子体诱变结合碘乙酸和脱氢表雄酮筛选获得的高DHA含量突变体的代谢分析

Metabolic Analysis of Mutants With High DHA Content Achieved With ARTP Mutagenesis Combined With Iodoacetic Acid and Dehydroepiandrosterone Screening.

作者信息

Zeng Lei, Bi Yanqi, Guo Pengfei, Bi Yali, Wang Tiantian, Dong Liang, Wang Fangzhong, Chen Lei, Zhang Weiwen

机构信息

Laboratory of Synthetic Microbiology, School of Chemical Engineering & Technology, Tianjin University, Tianjin, China.

Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, China.

出版信息

Front Bioeng Biotechnol. 2021 Nov 18;9:738052. doi: 10.3389/fbioe.2021.738052. eCollection 2021.

Abstract

High DHA production cost caused by low DHA titer and productivity of the current strains is a bottleneck for its application in competition with traditional fish-oil based approach. In this study, atmospheric and room-temperature plasma with iodoacetic acid and dehydroepiandrosterone screening led to three mutants, 6-8, 6-16 and 6-23 all with increased growth and DHA accumulations. A LC/MS metabolomic analysis revealed the increased metabolism in PPP and EMP as well as the decreased TCA cycle might be relevant to the increased growth and DHA biosynthesis in the mutants. Finally, the mutant 6-23, which achieved the highest growth and DHA accumulation among all mutants, was evaluated in a 5 L fermentor. The results showed that the DHA concentration and productivity in mutant 6-23 were 41.4 g/L and 430.7 mg/L/h in fermentation for 96 h, respectively, which is the highest reported so far in literature. The study provides a novel strain improvement strategy for DHA-producing .

摘要

当前菌株的二十二碳六烯酸(DHA)滴度和生产率较低,导致DHA生产成本高昂,这是其在与传统鱼油方法竞争应用中的一个瓶颈。在本研究中,通过大气室温等离子体结合碘乙酸和脱氢表雄酮筛选,获得了三个突变体6-8、6-16和6-23,它们的生长和DHA积累均有所增加。液相色谱/质谱代谢组学分析表明,磷酸戊糖途径(PPP)和糖酵解途径(EMP)代谢增加以及三羧酸循环(TCA)减少可能与突变体生长增加和DHA生物合成有关。最后,在5 L发酵罐中对在所有突变体中生长和DHA积累最高的突变体6-23进行了评估。结果表明,突变体6-23在96小时发酵中的DHA浓度和生产率分别为41.4 g/L和430.7 mg/L/h,这是迄今为止文献报道的最高值。该研究为DHA生产提供了一种新的菌株改良策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/272c/8637758/1a16a88d9567/fbioe-09-738052-g001.jpg

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