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用于生产聚(3-羟基丁酸酯-co-3-羟基戊酸酯)的重组大肠杆菌菌株的构建

Construction of recombinant Escherichia coli strains for production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate).

作者信息

Law Kin-Ho, Chan Pui-Ling, Lau Wai-Sum, Cheng Yin-Chung, Leung Yun-Chung, Lo Wai-Hung, Lawford Hugh, Yu Hoi-Fu

机构信息

Open Laboratory of Chirotechnology of the Institute of Molecular Technology for Drug Discovery & Synthesis, Applied Biology and Chemical Technology, Hong Kong Polytechnic University, Hong Kong, China.

出版信息

Appl Biochem Biotechnol. 2004 Spring;113-116:361-72.

Abstract

Plastic wastes constitute a worldwide environmental problem, and the demand for biodegradable plastics has become high. One of the most important characteristics of microbial polyesters is that they are thermoplastic with environmentally degradable properties. In this study, pUC19/PHA was cloned and transformed into three different Escherichia coli strains. Among the three strains that were successfully expressed in the production of polyhydroxyalkanoates (PHA), E. coli HMS174 had the highest yield in the production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) (P[HB-HV]). The cell dry weight and PHA content of recombinant HMS174 reached as high as 10.27 g/L and 43% (w/w), respectively, in fed-batch fermentor culture. The copolymer of PHA, P(HB-HV), was found in the cells, and the biopolymers accumulated were identified and analyzed by gas chromatography, proton nuclear magnetic resonance spectroscopy, and differential scanning calorimetry. We demonstrated clearly that the E. coli host for PHA production has to be carefully selected to obtain a high yield. The results obtained indicated that a superior E. coli with high PHA production can be constructed with a desirable ratio of P(HB-HV), which has potential applications in industry and medicine.

摘要

塑料垃圾构成了一个全球性的环境问题,因此对可生物降解塑料的需求变得很高。微生物聚酯最重要的特性之一是它们具有热塑性且具备环境可降解性能。在本研究中,pUC19/PHA被克隆并转化到三种不同的大肠杆菌菌株中。在成功表达聚羟基脂肪酸酯(PHA)的三种菌株中,大肠杆菌HMS174在聚(3-羟基丁酸酯-co-3-羟基戊酸酯)(P[HB-HV])的生产中产量最高。在补料分批发酵罐培养中,重组HMS174的细胞干重和PHA含量分别高达10.27 g/L和43%(w/w)。在细胞中发现了PHA的共聚物P(HB-HV),并通过气相色谱、质子核磁共振光谱和差示扫描量热法对积累的生物聚合物进行了鉴定和分析。我们清楚地证明,必须仔细选择用于PHA生产的大肠杆菌宿主以获得高产率。所得结果表明,可以构建出具有理想P(HB-HV)比例、高产PHA的优良大肠杆菌,其在工业和医学上具有潜在应用价值。

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