酵母能量偶联酶法合成Gln体系的建立及条件优化研究

Study on Establishment of Enzymatic Synthesis of Gln by Coupling Yeast-energy System and Optimization of Conditions

[目的]建立酵母能量偶联体系合成谷氨酰胺并优化体系反应条件。[方法]利用酵母下脚料培养工程菌乳糖诱导所得的谷氨酰胺合成酶,与酵母糖酵解产生的ATP能量偶联,酶法合成谷氨酰胺,并优化反应体系中各组分,提高谷氨酰胺产量。[结果]2%甲苯预处理干酵母;反应体系各组分的终浓度分别是NH4+100 mmol/L,Glu200 mmol/L,磷酸盐100 mmol/L,Mn2+10 mmol/L,Mg2+不另加;谷氨酸转化率达94.5%,谷氨酰胺产量为27.6 g/L。[结论]成功建立酵母能量偶联酶法合成谷氨酰胺体系,并对反应体系进行放大和初步优化,为酶法合成谷氨酰胺的工业化进程奠定了基础。

[Objective] To establish yeast energy-coupled system and optimize reaction conditions of the glutamine synthesis system.[Methods] Lactose induced wasted yeast cultured engineering bacteria to express glutamine synthetase,coupling ATP energy generated from yeast glycolysis,and catalyze enzymatic synthesis of L-glutamine.And optimize each component of the reaction system to improve L-glutamine production.[Results] Pretreatment of dry yeast with 2% toluene is appropriate.The final concentration of each component of the reaction system are NH4＋ 100 mmol/L,Glu 100 mmol/L,phosphate 100 mmol/L,Mn2＋ 10 mmol/L,Mg2＋ non plus.Moreover,the conversion rate of glutamate was 94.5%,and L-glutamine production was 27.6 g/L.[Conclusion] The yeast energy-coupled enzymatic synthesis of glutamine system was successfully established.We also enlarged and preliminary optimized the reaction system.Thus,the enzymatic synthesis of glutamine provides the basis for the process of industrialization.