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. 2021 Jan 25;37(1):196-206.
doi: 10.13345/j.cjb.200243.

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

[Article in Chinese]
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Free article

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

[Article in Chinese]
Xiangju Wei et al. Sheng Wu Gong Cheng Xue Bao. .
Free article

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.

聚羟基脂肪酸酯 (Polyhydroxyalkanoates,PHAs) 是一种具有优质生物相容性的可降解生物基材料,其理化性质优越,具备替代石油基塑料的潜力。P(3HB-co-LA) 是PHAs的一种,融合了聚乳酸 (Polylactic acid,PLA) 和聚3-羟基丁酸 (poly(3-hydroxybutyrate),P(3HB)) 共同的优点,具有更好的韧性和透明度。文中首先在大肠杆菌MG1655中通过质粒表达外源基因phaA、phaB、phaCm和pct(th),发酵生产出乳酸含量为23.8 mol%的P(3HB-co-LA),在此基础上缺失dld基因得到菌株WXJ01-03,其合成的聚合物中乳酸组分含量提升至37.2 mol%。当硫酯酶基因ydiI和yciA基因继续被敲除后,生产的共聚物中乳酸组分进一步提升至42.3 mol%和41.1 mol%。最后将3个基因dld、yciA和ydiI同时缺失得到重组菌株WXJ03-03,并通过该重组菌株获得了乳酸组分含量为46.1 mol%的共聚物。通过比较不同碳源的发酵结果得知,木糖有利于提高共聚物中乳酸组分含量。上述实验结果表明,在木糖发酵中短链硫酯酶基因缺失阻碍了大肠杆菌胞内的LA-CoA被降解,可有效提高聚合物中乳酸组分的摩尔百分比。.

Keywords: Escherichia coli; P(3HB-co-LA); polyhydroxyalkanoates; thioesterase; xylose fermentation.

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