使用葡萄糖和酸作为共同基质的持续食批量种植:在Pseudomonas sppp中微调聚氧甲酸单体成分的策略
Pedro Henrique Santos-Oliveira1, Jefferson Gonçalves Pinheiro Silva2, Lars Mathias Blank3
1Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil; iAMB - Institute of Applied Microbiology, ABBt - Aachen Biology and Biotechnology, RWTH Aachen University, Worringerweg 1, 52074 Aachen, Germany.
International journal of biological macromolecules
|November 23, 2023
概括
一个 伪虫 sp. 缺少二甲基异酸盐酶 (prpB) 的突变物能有效地产生多基酸盐 (PHA) 和可调节的奇链酸盐 (HA). 这为各种应用提供了一种微调PHA成分的方法.
科学领域:
- 微生物的新陈代谢
- 生物聚合物生产生产
- 合成生物学 合成生物学
背景情况:
- 伪omonas sp. 这种病毒是假的. LFM693 具有 2 甲基酸盐循环用于酸盐氧化,涉及 2 甲基酸盐酶 (prpB).
- 当与和葡萄糖一起供应时,prpB的破坏导致突变能够产生多甲基酸盐 (PHA) 与偶数和奇数链长甲基酸盐 (HA) 的突变.
研究的目的:
- 为了研究中链长的多基酸盐 (mclPHA) 的产生,在Pseudomonas sp.的prpB受损突变体中进行研究. 在 LFM693.3.
- 通过操纵养策略来控制和微调mclPHA的组成,特别是HA的结合.
主要方法:
- 使用恒定料批量配置来控制PHA组成并减轻酸毒性.
- 在食溶液中变化了酸盐/葡萄糖的摩尔比率,以影响HAodd的结合.
- 分析了PHA含量,分子组成和转化因子.
主要成果:
- 从7%到30%实现了HAodd的线性整合,与酸/葡萄糖比率直接相关.
- 获得了平均PHA含量为52%的细胞干重,优于C8,C9和C10的3-基甲酸盐.
- 观察到酸到HAodd转换因子 (YHAodd/prop) 之间的0.36和0.53mol·mol-1,低于理论最大值.
结论:
- 这种prpB破坏的突变使得mclPHA分子组成可以在最小的过程控制下进行微调.
- 观察到的转化效率和缺乏二甲基异酸盐副产品支持prpB-突变物继续氧化酸盐的能力.
- 这项研究表明,通过代谢工程和受控发酵,为特定应用量身定制PHA共聚物的可行策略.
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