双酶偶联生物合成L-酪氨酸  

Biosynthesis of L-Tyrosine with Whole Cell Coupled Aspartate Aminotransferase and Tyrosine Phenol-Lyase

作　　者：冯莹莹[1] 刘均忠[1] 张宏娟[2] 刘茜[1] 焦庆才[1] 

机构地区：[1]南京大学生命科学学院医药生物技术国家重点实验室,江苏南京210093 [2]南京医科大学药学院,江苏南京210029

出　　处：《精细化工》2014年第5期570-574,606,共6页Fine Chemicals

基　　金：国家自然科学基金青年科学基金项目(21302100)~~

摘　　要：多酶偶联催化是酶法制备手性药物中间体的重要方法之一。该文采用双酶偶联体系,利用天冬氨酸转氨酶全细胞催化L-天冬氨酸转氨至苯丙酮酸,生成L-苯丙氨酸,同时得到中间产物丙酮酸;反应体系中的酪氨酸酚裂解酶全细胞催化丙酮酸、苯酚和氨酶法合成L-酪氨酸。经考察确定了双酶偶联反应的最佳条件为:温度40℃,pH=8.5,底物苯丙酮酸质量浓度为25 g/L,苯丙酮酸与L-天冬氨酸摩尔比1∶1.2,天冬氨酸转氨酶与酪氨酸酚裂解酶细胞质量比1∶1,4 mmol/L PLP,0.1 g/L吐温80。30 g/L的氯化铵对双酶偶联反应有促进作用。双菌双酶偶联生物法合成L-酪氨酸,充分利用了反应副产物丙酮酸得到附加值较高的产品,对资源合理利用及绿色合成工艺具有参考意义。Multienzyme coupled catalysis is one of the important methods in the enzymatic preparation of chiral drug intermediates. In this study, the ammonia of L-aspartic acid was transferred to the phenylpyruvic acid, and L-phenylalanine with byproduct pyruvic acid was synthesized by aspartate aminotransferase （AspAT） whole cell. Then, L-tyrosine was enzymatically synthesized when the substrate phenol and tyrosine phenol-lyase（TPL） whole cell were added into the reaction system. Some factors affecting the dual enzymatic catalysis were investigated. The optimal conditions were 40 ℃, pH = 8.5,25 g/L of phenylpyruvic acid（PPA）, n（PPA） ：rt（L-Asp） = 1 ： 1.2, m（AspAT） ：m（TPL） = 1：1, 4 mmol/L of PLP,0. 1 g/L of Tween 80. 30 g/L of ammonium chloride had a promoting effect on the dual enzyme coupled catalysis reaction. The method of biological synthesis of L-tyrosine with coupled dual enzyme in dual host not only makes full use of the by-product of the reaction pyruvie acid for higher value-added products, but has reference significance for the reasonable utilization of resources and green synthesis process.

关 键 词：双酶偶联 天冬氨酸转氨酶 酪氨酸酚裂解酶 L-酪氨酸 生物工程 

分 类 号：TQ464.7[化学工程—制药化工]