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用Pseudomonas putida KT2440的进化导向耐受性工程来生产航空燃料前体异醇

Hyun Gyu Lim¹, Aparajitha Srinivasan², Russel Menchavez²

¹Department of Bioengineering, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA; Joint BioEnergy Institute, 5885 Hollis street, 4th floor, Emeryville, CA 94608, USA; Department of Biological Sciences and Bioengineering, Inha University, Inha-ro 100, Michuhol-gu, Incheon 22212, Korea.

Metabolic engineering

|May 21, 2025

概括

耐受性适应性实验室进化 (TALE) 创造了 Pseudomonas putida 菌株,更耐受生物燃料前体异二醇. 将TALE与基因工程结合起来,提高了这种有价值的化学物质的微生物生产.

科学领域:

微生物生物技术微生物生物技术
合成生物学合成生物学
生物化学工程是生物化学工程.

背景情况:

异二醇是航空燃料和化学品的关键前体,可从可再生资源中生产.
目前Pseudomonas putida中的微生物对异二醇的生产受到毒性限制,阻碍了其商业可行性.
适应性实验室进化策略对于开发强大的微生物宿主至关重要.

研究的目的:

使用耐受性适应性实验室进化 (TALE) 来提高P. putida的异二醇耐受性和产量.
通过全基因组再测序来确定赋予异二醇耐受性的遗传突变.
为了改造P. putida菌株以改善生物燃料前体的合成.

主要方法:

开展了一项关于野生型和改性P. putida菌株的TALE活动,进化独立的血统.
进行全基因组再测序,以确定进化菌株中的融合突变.
利用突变区域的逆向工程和蛋白质学引导的基因删除 (fleQ) 来验证耐受性机制.

主要成果:

进化的P. putida克隆在高异二醇度 (8 g/L) 时表现出显著的增长,与父菌株不同.
在特定的遗传区域 (gnuR,ttgB-PP_1394,PP_3024-PP_5558,PP_1695) 发现了融合突变.
在一个进化的克隆中删除fleQ解决了耐受性-生产权衡,恢复了异二醇位.

结论:

TALE是一种有效的策略,用于生成具有增强异二醇耐受性的P. putida菌株.
实验室进化和理性工程的综合方法克服了微生物生物燃料生产中的产品毒性.
开发出强大的宿主可以最大限度地降低产品的毒性,为商业上可行的异二醇合成铺平了道路.

关键词:

3-甲基-3--1--1-醇的使用情况.适应性实验室进化的演变.伪omonas putida可以使用.这是一种异二醇 (isoprenol).可持续发展的航空燃料 (SAF)耐受性工程是一种耐受性工程.

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