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[短链硫酯酶缺陷对大肠杆菌中聚(3-羟基丁酸酯-co-乳酸)生物合成的影响]

[Effect of short-chain thioesterase deficiency on P(3HB-co-LA) biosynthesis in Escherichia coli].

作者信息

Wei Xiangju, Wu Ju, Guo Pengye, Zhou Shengmin, Wu Hui

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2021 Jan 25;37(1):196-206. doi: 10.13345/j.cjb.200243.

Abstract

Polyhydroxyalkanoates (PHAs) have obtained much attention in biomaterial fields due to their similar physicochemical properties to those of the petroleum-derived plastics. Poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)] is one member of the PHAs family, and has better toughness and transparency compared to existing polylactic acid (PLA) and poly[(R)-3-hydroxybutyrate] [P(3HB)]. First, we confirmed the one-step biosynthesis of P(LA-co-3HB) with the lactate fraction of 23.8 mol% by introducing P(3HB-co-LA) production module into Escherichia coli MG1655. Then, the lactate fraction was increased to 37.2 mol% in the dld deficient strain WXJ01-03. The genes encoding the thioesterases, ydiI and yciA, were further knocked out, and the lactate fraction in the P(3HB-co-LA) was improved to 42.3 mol% and 41.1 mol% respectively. Strain WXJ03-03 with dld, ydiI and yciA deficient was used for the production of the LA-enriched polymer, and the lactate fraction was improved to 46.1 mol%. Notably, the lactate fraction in P(3HB-co-LA) from xylose was remarkably higher than from glucose, indicating xylose as a potent carbon source for P(3HB-co-LA) production. Therefore, the deficiency of thioesterase may be considered as an effective strategy to improve the lactate fraction in P(3HB-co-LA) in xylose fermentation.

摘要

聚羟基脂肪酸酯(PHA)因其与石油衍生塑料相似的物理化学性质而在生物材料领域备受关注。聚(3-羟基丁酸酯-co-乳酸酯)[P(3HB-co-LA)]是PHA家族的一员,与现有的聚乳酸(PLA)和聚(R)-3-羟基丁酸酯[P(3HB)]相比,具有更好的韧性和透明度。首先,通过将P(3HB-co-LA)生产模块引入大肠杆菌MG1655,我们证实了一步法生物合成乳酸分数为23.8 mol%的P(LA-co-3HB)。然后,在dld缺陷菌株WXJ01-03中,乳酸分数增加到37.2 mol%。进一步敲除编码硫酯酶ydiI和yciA的基因,P(3HB-co-LA)中的乳酸分数分别提高到42.3 mol%和41.1 mol%。使用dld、ydiI和yciA缺陷的菌株WXJ03-03生产富含LA的聚合物,乳酸分数提高到46.1 mol%。值得注意的是,木糖来源的P(3HB-co-LA)中的乳酸分数显著高于葡萄糖来源的,表明木糖是P(3HB-co-LA)生产的有效碳源。因此,硫酯酶缺陷可被视为提高木糖发酵中P(3HB-co-LA)乳酸分数的有效策略。

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