过量表达苹果酸酶对E.coli脂肪酸合成能力的影响
详细信息Effect of overexpression of malic enzyme on fatty acid production of Escherichia coli
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摘要: 为了考察苹果酸酶对原核生物脂肪酸合成能力的影响,本研究从E.coliK-12中克隆了苹果酸酶基因（NADPME,EC1.1.1.40）MaeB,并插入质粒pET30a-T,构建了重组质粒pMX20,经IPTG诱导在E.coli BL2（IDE3）获得了大量表达。摇瓶发酵结果表明,过量表达苹果酸酶基因会改变大肠杆菌脂肪酸合成能力,并在添加适宜底物苹果酸（15mmol/L）时,胞内总酯含量比受体菌提高了4倍,约197.74mg/g,产率为1.89%。本研究为脂肪酸生产提供了优良菌株,具有一定的应用开发前景。Abstract: In order to investigate the impact of malic enzyme on the capacity of fatty acid synthesis about prokaryotes, the MaeB gene encoding NADP ME was amplified from E.coli K-12 genome by PCR and inserted in the vector pET30a-T to build recombinanted plasmid pMX20. Malic enzyme was overproduced by E.coli BL21 (DE3) harboring pMX20 under the condition of IPTG induction. The results of fermentation indicated that overexpression of malic enzyme in E.coli would have influence on the capacity of fatty acid synthesis, and meanwhile, the total lipids in recombinanted cell was 4 times to E.coli BL21 (DE3) under the condition of adding appropriate substrate of malic acid (about 197.74mg/g) , and the yield was 1.89%. These results also showed that the recombinanted strain MX20 was useful and had certain development prospects which would provide an excellent strain for fatty acid production.
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Keywords:
- E. coli;malic enzyme;fatty acid; /
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