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首页> 外文期刊>Applied and Environmental Microbiology >Cloning of the Alcaligenes latus Polyhydroxyalkanoate Biosynthesis Genes and Use of These Genes for Enhanced Production of Poly(3-hydroxybutyrate) in Escherichia coli
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Cloning of the Alcaligenes latus Polyhydroxyalkanoate Biosynthesis Genes and Use of These Genes for Enhanced Production of Poly(3-hydroxybutyrate) in Escherichia coli

机译：产碱菌聚羟基链烷酸酯生物合成基因的克隆及这些基因在大肠杆菌中增强聚（3-羟基丁酸酯）生产的用途

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Polyhydroxyalkanoates (PHAs) are microbial polyesters that can be used as completely biodegradable polymers, but the high production cost prevents their use in a wide range of applications. RecombinantEscherichia coli strains harboring the Ralstonia eutropha PHA biosynthesis genes have been reported to have several advantages as PHA producers compared with wild-type PHA-producing bacteria. However, the PHA productivity (amount of PHA produced per unit volume per unit time) obtained with these recombinantE. coli strains has been lower than that obtained with the wild-type bacterium Alcaligenes latus. To endow the potentially superior PHA biosynthetic machinery to E. coli, we cloned the PHA biosynthesis genes from A. latus. The three PHA biosynthesis genes formed an operon with the order PHA synthase, β-ketothiolase, and reductase genes and were constitutively expressed from the natural promoter in E. coli. RecombinantE. coli strains harboring the A. latus PHA biosynthesis genes accumulated poly(3-hydroxybutyrate) (PHB), a model PHA product, more efficiently than those harboring the R. eutropha genes. With a pH-stat fed-batch culture of recombinantE. coli harboring a stable plasmid containing the A. latus PHA biosynthesis genes, final cell and PHB concentrations of 194.1 and 141.6 g/liter, respectively, were obtained, resulting in a high productivity of 4.63 g of PHB/liter/h. This improvement should allow recombinant E. coli to be used for the production of PHB with a high level of economic competitiveness.
机译：聚羟基链烷酸酯（PHA）是可以用作完全可生物降解的聚合物的微生物聚酯，但是高昂的生产成本使其无法在广泛的应用中使用。据报道，与野生型生产PHA的细菌相比，带有富营养的Ralstonia eutropha PHA生物合成基因的重组大肠杆菌菌株具有许多优势。然而，用这些重组体E获得的PHA生产率（每单位时间每单位体积产生的PHA量）。大肠埃希菌菌株低于野生型细菌产碱杆菌。为了赋予潜在的优于大肠杆菌的PHA生物合成机制，我们从A. latus克隆了PHA生物合成基因。这三个PHA生物合成基因与PHA合成酶，β-酮硫解酶和还原酶顺序形成操纵子，并从大肠杆菌中的天然启动子组成性表达。重组E.携带A. latus PHA生物合成基因的大肠杆菌菌株比带有R. eutropha基因的菌株更有效地积累了模型PHA产物聚（3-羟基丁酸酯）（PHB）。用pH调节器补料分批培养重组E.含有稳定的质粒的大肠埃希菌含有La.Atus PHA生物合成基因，最终细胞和PHB的浓度分别为194.1和141.6 g / L，从而实现了4.63 g PHB / L / h的高生产率。这种改进应使重组大肠杆菌可用于生产具有高水平经济竞争力的PHB。

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