Establishment of a metabolic pathway to introduce the 3-hydroxyhexanoate unit into LA-based polyesters via a reverse reaction of β-oxidation in Escherichia coli LS5218
Establishment of a metabolic pathway to introduce the 3-hydroxyhexanoate unit into LA-based polyesters via a reverse reaction of β-oxidation in Escherichia coli LS5218
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DOI：
10.1016/j.polymdegradstab.2010.01.029
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发表时间：
2010-08-01
影响因子：
5.9
通讯作者：
Taguchi, Seiichi
中科院分区：
文献类型：
--
作者：
Shozui, Fumi;Matsumoto, Ken'ichiro;Taguchi, Seiichi
在大肠杆菌LS 5218中构建了一种新的乳酸（LA）-羟基丁酸（3 HB）-羟基己酸（3 HHx）共聚物，该共聚物含有3个编码LA聚合酶（LPE）、丙酰辅酶A（CoA）转移酶（PCT）和烯酰辅酶A水合酶（PhaJ 4）的基因。当重组LS 5218在葡萄糖上生长并以丁酸盐进料时，可能通过β-氧化途径和PhaJ 4的逆反应提供3 HB-CoA和3 HHx-CoA。LPE将两种单体3 HB-CoA和3 HHx-CoA与由PCT产生的LA-CoA共聚，以产生三元共聚物。气相色谱分析表明，三元共聚物由2.7-34摩尔%的LA、38-81摩尔%的3 HB和17-33摩尔%的3 HHx单元组成，其可以根据培养基中进料的丁酸盐浓度而变化。此外，H-1-C-13 COSY NMR分析提供了聚合物中LA和3 HHx单元之间连接的证据。(C)2010爱思唯尔有限公司保留所有权利。
New lactate (LA)-based terpolymers, P[LA-co-3-hydroxybutyrate (3HB)-co-3-hydroxyhexananoate (3HHx)]s, were produced in recombinant Escherichia coli LS5218 harboring three genes encoding LA-polymerizing enzyme (LPE), propionyl-coenzyme A (CoA) transferase (PCT) and (R)-specific enoyl-CoA hydratase (PhaJ4). When the recombinant LS5218 was grown on glucose with the feeding of butyrate, 3HB-CoA and 3HHx-CoA were supplied, probably via reverse reactions of the beta-oxidation pathway and PhaJ4. LPE copolymerized the two monomers 3HB-CoA and 3HHx-CoA with LA-CoA, which was generated by PCT, to yield the terpolymers. Gas chromatography analysis revealed that the terpolymers consisted of 2.7-34 mol% LA, 38-81 mol% 3HB and 17-33 mol% 3HHx units, which can be varied depending on the butyrate concentration fed in the medium. In addition, H-1-C-13 COSY NMR analysis provided evidence for a linkage between LA and 3HHx units in the polymer. (C) 2010 Elsevier Ltd. All rights reserved.