生物法合成胍基丁胺具有成本低、绿色高效等优点,开发微生物发酵法产精氨酸脱羧酶,利用该酶催化精氨酸生成胍基丁胺具有重要意义。该实验以重组大肠杆菌为研究对象,进行单因素实验考察了不同诱导培养温度对菌体细胞表达重组蛋白的影响,不同pH条件对精氨酸脱羧酶活性的影响。然后采用实验设计优化培养基组成,分析各种成分对重组蛋白生产的影响。通过将抑制性成分去除,对蛋白表达有促进作用成分加量,使得精氨酸脱羧酶的酶活提高了2.23倍。优化了转化过程中辅酶的添加量,降低了胍基丁胺的生产成本。15 L发酵罐发酵与转化放大实验,硫酸胍基丁胺产量达到了279.21 g/L,转化率98%。该研究为胍基丁胺的工业化生产奠定了基础。
An approach to produce agmatine from arginine using arginine decarboxylase was developed. An arginine decarboxylase was expressed in Escherichia coli and the condition for enzyme expression was optimized. The activity of the enzyme increased 2.23 times by removing the inhibitive components and increasing the amount of those that promote protein expression. The effect of coenzyme on the yield of agmatine in the transformation process was also optimized. In pilot-scale in 15 L fermenter, the yield of agmatine reached 279.21 g/L with conversion rate of 98%.
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