[Construction of recombinant Escherichia coli strains producing poly (4-hydroxybutyric acid) homopolyester from glucose].
[Construction of recombinant Escherichia coli strains producing poly (4-hydroxybutyric acid) homopolyester from glucose].
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发表时间：
2005-06
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通讯作者：
Shuishan Song;Hong-Lin Ma;Zhen-xian Gao;Zhen-hua Jia;Xia Zhang
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作者：
Shuishan Song;Hong-Lin Ma;Zhen-xian Gao;Zhen-hua Jia;Xia Zhang
本研究旨在构建以葡萄糖为唯一碳源生产聚4-羟基丁酸均聚酯的重组大肠杆菌菌株。来自大肠杆菌的谷氨酸：琥珀酸半醛转氨酶基因、来自大肠杆菌的谷氨酸脱羧酶基因。coli中的PHA合成酶基因和真养罗尔斯通氏菌（Ralstonia eutropha）的4-羟基丁酸脱氢酶基因，并将其组装到含有真养罗尔斯通氏菌（Ralstonia eutropha）的PHA合成酶基因和克鲁弗梭菌（Clostridium kluyveri）的4-羟基丁酸：CoA转移酶基因的质粒pKSSE 5.3上。转化E.大肠杆菌中，建立了以葡萄糖为原料经TCA循环中间产物α-酮戊二酸合成聚4-羟基丁酸的途径。大肠杆菌菌株。重组菌在以葡萄糖为碳源的Luria-Bertani发酵液中培养时，可合成均聚酯P（4 HB）。在以葡萄糖为唯一碳源的无机盐M9培养基中，分别添加酵母膏、胰蛋白胨、水解酪蛋白，可使P（4 HB）积累量提高到细胞干重的30%。
The aim of this study is to construct recombinant Escherichia coli strains capable of producing poly(4-hydroxybutyric acid) homopolyester from glucose as sole carbon source. A glutamate: succinate semialdehyde transaminase gene from Escherichia coli, a glutamate decarboxylase gene from E. coli, and a 4-hydroxybutyrate dehydrogenase gene from Ralstonia eutropha were cloned by PCR and assembled onto the plasmid pKSSE5.3 which haboured the PHA synthase gene from Ralstonia eutropha and 4-hydroxybutyrate: CoA transferase from Clostridium kluyveri. The resulting plasmids were transformed into E. coli and the pathway for biosynthesis of poly(4-hydroxybutyric acid) from glucose via alpha-ketoglutarate, an intermediate in TCA cycle was established in recombinant E. coli strains. Recombinant strains synthesized the homopolyester P(4HB), when cells were cultivated in Luria-Bertani broth with glucose as carbon source. P(4HB) accumulation was enhanced up to 30% of cell dry weight, when cells were cultivated in mineral salts M9 medium plus glucose as sole carbon source with addition of yeast extract, tryptone, casein hydrolate into medium respectively.