Biosynthesis of novel lactate-based polymers containing medium-chain-length 3-hydroxyalkanoates by recombinant Escherichia coli strains from glucose
Biosynthesis of novel lactate-based polymers containing medium-chain-length 3-hydroxyalkanoates by recombinant Escherichia coli strains from glucose
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DOI：
10.1016/j.jbiosc.2019.01.009
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发表时间：
2019-08-01
影响因子：
2.8
通讯作者：
Matsusaki, Hiromi
中科院分区：
文献类型：
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作者：
Goto, Saki;Hokamura, Ayaka;Matsusaki, Hiromi
以葡萄糖为唯一碳源，在FadR缺陷的大肠杆菌中合成了含有中链3-羟基烷酸酯(MCL-3HA)的新型乳酸(LA)聚合物。编码LA和3-羟基丁酸酯(3HB)的编码单体通过脂肪酸生物合成途径提供酶[丙酰辅酶A转移酶(PCT)、D-乳酸脱氢酶(DLDH)、β-酮硫醇酶(PHAA)和依赖于NADPH的乙酰乙酰辅酶A还原酶(PHAB)]、MCL-3HA单体提供酶[(R)-3-羟基酰基-ACP硫酯酶(PHAG)和(R)-3-羟基酰基(3HA)-辅酶A连接酶]，以及假单胞菌修饰的聚羟基烷酸(PHA)合成酶[PhaC1(STQK)]。61-3导入到大肠杆菌LS5218中。合成了一种新型的LA基共聚物P(LA-co-3HB-co-3HA)。核磁共振氢谱分析表明，P(LA-co-3HB-co-3HA)的组成为19.7摩尔%LA(C-3)、74.9摩尔%3HB(C-4)和5.4摩尔%MCL-3HA单元C-8和C-10。此外，携带这些基因的重组大肠杆菌CAG18497(不包括phaAB基因)积累了P(92.0%LA-co-3HA)，其新的单体组成包含C-3、C-8、C-10和C-12。~(13)C-核磁共振分析表明聚合物中存在LA-3HA序列。P(92.0%LA-co-3HA)的溶剂浇铸膜具有与聚乳酸相似的透明性。(C)2019年，日本生物技术学会。版权所有。
Novel lactate (LA)-based polymers containing medium-chain-length 3-hydroxyalkanoates (MCL-3HA) were produced in fadR-deficient Escherichia coli strains from glucose as the sole carbon source. The genes encoding LA and 3-hydroxybutyrate (3HB) monomers supplying enzymes [propionyl-CoA transferase (PCT), D-lactate dehydrogenase (DLDH), beta-ketothiolase (PhaA), and NADPH-dependent acetoacetyl-CoA reductase (PhaB)], MCL-3HA monomers supplying enzymes [(R)-3-hydroxyacyl-ACP thioesterase (PhaG) and (R)-3-hydroxyacyl (3HA)-CoA ligase] via fatty acid biosynthesis pathway, and modified polyhydroxyalkanoate (PHA) synthase [PhaC1(STQK)] of Pseudomonas sp. 61-3 were introduced into E. coli LS5218. This resulted in the synthesis of a novel LA-based copolymer, P(LA-co-3HB-co-3HA). H-1-nuclear magnetic resonance (NMR) analysis revealed the composition of P(LA-co-3HB-co-3HA) to be 19.7 mol% LA (C-3), 74.9 mol% 3HB (C-4), and 5.4 mol% MCL-3HA units of C-8 and C-10. Furthermore, the recombinant E. coli CAG18497 strain carrying these genes, excluding the phaAB genes, accumulated P(92.0% LA-co-3HA) with a novel monomer composition containing C-3, C-8, C-10, and C-12. C-13-NMR analysis showed the existence of LA-3HA sequence in the polymer. The solvent cast film of P(92.0% LA-co-3HA) exhibited transparency similar to poly(lactic acid). (C) 2019, The Society for Biotechnology, Japan. All rights reserved.