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Method for promoting acetylglucosamine synthesis of bacillus subtilis

机译:促进乙酰葡糖胺合成枯草芽孢杆菌的方法

摘要

The present invention relates to a method for promoting acetylglucosamine synthesis of Bacillus subtilis, which belongs to the field of genetic engineering. The present invention adopts the recombinant Bacillus subtilis BSGNKAP2 as a starting strain, exogenously introducing pyruvate carboxylase BalpycA derived from Bacillus cereus, eliminating the central carbon metabolism overflow of the Bacillus subtilis and avoiding the synthesis of the by-product acetoin; further, five exogenous reducing force metabolic reactions are introduced to replace the reaction of generating NADH in glycolysis pathway and tricarboxylic acid cycle to reconstruct intracellular reducing force metabolism, which specifically comprise glyceraldehyde-3-phosphate ferredoxin dehydrogenase, isocitrate NAD+ dehydrogenase, a malate quinone dehydrogenase, a ketoacid ferredoxin oxidoreductase and a nitrogenase ferritin. In a shake-flask fermentation process using a complex medium, acetylglucosamine yield of the recombinant strain BSGNKAP8 is 24.50 g/L, acetylglucosamine/glucose yield is 0.469 g/g, respectively 1.97 times and 2.13 times of those of the starting strain BSGNKAP2.
机译:本发明涉及促进甲虫淀粉胺合成的植物枯草芽孢杆菌的方法,该方法属于基因工程领域。本发明采用重组芽孢杆菌BSGNKAP2作为起始菌株,外源引入衍生自芽孢杆菌的丙酮酸羧酸甲酶BalpyCA,消除了枯草芽孢杆菌的中央碳代谢溢出,避免了副产物的丙酮蛋白的合成;此外,引入了五种外源降低力代谢反应以取代在糖酵解途径和三羧酸循环中产生NADH的反应,以重建细胞内降低力代谢,其特异性包含甘氨酸醌脱氢酶的甘氨酸NAD +脱氢酶的细胞内降低力代谢。 ,酮酸芳米嗪氧化还原酶和氮酶铁蛋白。在使用复合介质的摇瓶发酵过程中,重组菌株Bsgnkap8的乙酰葡糖胺产率为24.50g / L,乙酰葡糖胺/葡萄糖产率为0.469g / g,分别为0.469g / g,分别为起始菌株Bsgnkap2的1.97倍和2.13倍。

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