基因工程法合成间苯三酚

发布时间：2018-06-11 11:14

本文选题：间苯三酚 + MarA　；参考：《生物技术通报》2017年12期

【摘要】：为进一步提高生物法合成间苯三酚的效率,探究生物合成间苯三酚的分子机理以及更加高效的发酵条件和培养基成分。对来自荧光假单胞菌(Pseudomonas fluorescens)的phl D基因进行了克隆,在BL21(DE3)中重建间苯三酚的胞内合成途径,超表达多重抗药基因正向调节因子(mar A)和乙酰辅酶A羧化酶(ACCase),并分析不同碳源及培养基离子环境对间苯三酚积累量的影响。结果显示,成功构建间苯三酚胞内合成途径,产量达到450 mg/L,mar A基因能够提高工程菌对间苯三酚抵抗力,使间苯三酚产量提高到1 060 mg/L;ACCase能够提高胞内丙二酰辅酶A的含量,使间苯三酚产量提高到2 120 mg/L;葡萄糖、甘油和丙酮酸钠三种碳源的比较中,葡萄糖对合成间苯三酚最为有利,分别是甘油和丙酮酸钠的1.8倍和2.4倍;培养基中钙镁离子对间苯三酚的合成没有影响。mar A和ACCase基因的超表达能大大提高间苯三酚生物合成的效率,以葡萄糖为碳源的无钙镁培养基为间苯三酚发酵的最适培养基。
[Abstract]:In order to further improve the efficiency of biosynthesis of benzol three, the molecular mechanism of biosynthesis of benzol and the more efficient fermentation conditions and medium components were explored. The Phl D gene from Pseudomonas fluorescens (Pseudomonas fluorescens) was cloned and the intracellular synthesis pathway was reconstructed in BL21 (DE3), and the overexpression was overexpressed in BL21 (DE3). The effect of multiple anti drug gene positive regulator (Mar A) and acetyl coenzyme A carboxylase (ACCase), and the effects of different carbon sources and medium ion environment on the accumulation of benzol three were analyzed. The results showed that the intracellular synthesis pathway of phenylephol was successfully constructed, the yield reached 450 mg/L, and mar A gene could improve the resistance of engineering bacteria to the phenylephenol three. The yield of three phenol is increased to 1060 mg/L; ACCase can increase the content of intracellular A two acyl coenzyme and increase the yield of interphenolol to 2120 mg/L; in the comparison of three carbon sources of glucose, glycerin and sodium pyruvate, glucose is most favorable to the synthesis of interphenols, 1.8 times and 2.4 times of glycerol and sodium propionate, respectively. The synthesis of benzol three has no effect on the overexpression of.Mar A and ACCase gene, which can greatly improve the efficiency of biosynthesis of phenolol. The non calcium magnesium medium with glucose as the carbon source is the most suitable medium for the fermentation of phenylene three.
【作者单位】：潍坊科技学院化工与环境学院;寿光富康制药研发部;
【基金】：海洋生物医药间苯三酚等制剂国际化发展能力建设项目[鲁财建指2014(No.18)]
【分类号】：TQ243.12

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