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在非中和条件下,影响D-乳酸生产的因素由Saccharomyces cerevisiae在非中和条件下

Dianti Rahmasari¹, Prihardi Kahar^1,2,3, Arthur Vinícius de Oliveira¹

¹Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe 658-8501, Hyogo, Japan.

|March 27, 2025

概括

在酵母中整合基因产生D-乳酸 (D-LA) 是生物塑料的关键. 这项研究发现,将D-LDH基因集成到PDC1位点产生了最高的D-LA (51g/L) 并改善了酵母花.

科学领域:

代谢工程是代谢工程.
合成生物学合成生物学
生物技术是生物技术.

背景情况:

D-乳酸 (D-LA) 是生物塑料和制药的重要组成部分.
用Saccharomyces cerevisiae进行D-LA生产的代谢工程通常涉及整合异质基因.
关于特定的基因组整合部位如何影响D-LA产量和酵母花的行为,目前的研究有限.

研究的目的:

为了研究将Leuconostoc pseudomesenteroides的编码子优化的D-LDH基因集成到Saccharomyces cerevisiae的各种基因组位置的影响.
评估不同整合场所对D-LA生产效率和副产品形成的影响.
分析基因组整合位点,基因表达和酵母菌化特性之间的相关性.

主要方法:

用LpDLDH基因集成到CYB2,PDC1,MPC1,PDC6,ADH1和PDC5位点中的Saccharomyces cerevisiae菌株的构建.
使用分析方法量化D-LA和乙醇标位.
基因表达分析与花相关的基因,应激适应和细胞壁完整性基因.
评价酵母菌的花性质.

主要成果:

在PDC1位点的LpDLDH集成导致了最高的D-LA标位 (51g/L) 与最小的乙醇.
在CYB2位点的整合产生了31.92g/L的D-LA,而MPC1和PDC6位点产生了较低的标位.
在ADH1和PDC5位点的整合未能产生D-LA,并导致完全失去了花.
观察到显著上调的花 (FLO5),应激适应 (HSP150),和细胞壁完整性 (YGP1,SED1,SCW11) 基因.
结合CYB2的菌株表现出强烈的花,表明强大的强度.

结论:

基因组位置选择对Saccharomyces cerevisiae中D-LA生产的代谢流量产生了关键的影响.
整合部位影响与细胞强度相关的基因表达,包括花和应激适应.
优化集成站点可以增强D-LA生产,并利用有益的酵母表型,如花,以提高工艺性能.

关键词:

D-乳酸是什么意思 D-乳酸是什么意思这种植物是Saccharomyces cerevisiae.发花过程中的花.基因转录基因转录集成的位置集成的位置

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