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通过代谢工程改造的大肠杆菌生物合成的聚(乳酸-共-3-羟基丁酸酯)中乳酸组分的增强。

Enhancement of lactate fraction in poly(lactate-co-3-hydroxybutyrate) biosynthesized by metabolically engineered E. coli.

作者信息

Zhang Binghao, Guo Pengye, Sun Xinye, Shang Yanzhe, Luo Yuanchan, Wu Hui

机构信息

State Key Laboratory of Bioreactor Engineering, Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism, School of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.

MOE Key Laboratory of Bio-Intelligent Manufacturing, School of Bioengineering, Dalian University of Technology, Dalian, China.

出版信息

Bioresour Bioprocess. 2024 Sep 19;11(1):88. doi: 10.1186/s40643-024-00803-2.

Abstract

Poly(lactate-co-3-hydroxybutyrate) [P(LA-co-3HB)] is a high-molecular-weight biomaterial with excellent biocompatibility and biodegradability. In this study, the properties of P(LA-co-3HB) were examined and found to be affected by its lactate fraction. The efficiency of lactyl-CoA biosynthesis from intracellular lactate significantly affected the microbial synthesis of P(LA-co-3HB). Two CoA transferases from Anaerotignum lactatifermentans and Bacillota bacterium were selected for use in copolymer biosynthesis from 11 candidates. We found that cot enhanced the lactate fraction by 31.56% compared to that of the frequently used modified form of propionyl-CoA transferase from Anaerotignum propionicum. In addition, utilizing xylose as a favorable carbon source and blocking the lactate degradation pathway further enhanced the lactate fraction to 30.42 mol% and 52.84 mol%, respectively. Furthermore, when a 5 L bioreactor was used for fermentation utilizing xylose as a carbon source, the engineered strain produced 60.60 wt% P(46.40 mol% LA-co-3HB), which was similar to the results of our flask experiments. Our results indicate that the application of new CoA transferases has great potential for the biosynthesis of other lactate-based copolymers.

摘要

聚(乳酸 - 共 - 3 - 羟基丁酸酯)[P(LA - co - 3HB)]是一种具有优异生物相容性和生物降解性的高分子量生物材料。在本研究中,对P(LA - co - 3HB)的性能进行了检测,发现其受乳酸组分的影响。细胞内乳酸生物合成乳酰辅酶A的效率显著影响了P(LA - co - 3HB)的微生物合成。从11种候选物中选择了来自乳酸发酵厌氧杆菌和芽孢杆菌属细菌的两种辅酶A转移酶用于共聚物生物合成。我们发现,与常用的丙酸厌氧杆菌的丙酰辅酶A转移酶修饰形式相比,cot使乳酸组分提高了31.56%。此外,利用木糖作为良好的碳源并阻断乳酸降解途径,可进一步将乳酸组分分别提高到30.42摩尔%和52.84摩尔%。此外,当使用5升生物反应器以木糖作为碳源进行发酵时,工程菌株产生了60.60 wt%的P(46.40摩尔% LA - co - 3HB),这与我们摇瓶实验的结果相似。我们的结果表明,新型辅酶A转移酶的应用在其他基于乳酸的共聚物生物合成方面具有巨大潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f55a/11413402/88899b71c667/40643_2024_803_Fig1_HTML.jpg

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