重组酪氨酸酚裂解酶全细胞催化合成L-酪氨酸  

Synthesis of L-Tyrosine by Whole Cell with Recombinant Tyrosine Phenol Lyase

作　　者：常俊俊[1] 刘均忠[1] 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

出　　处：《精细化工》2013年第10期1112-1116,1137,共6页Fine Chemicals

基　　金：国家技术创新基金(02CJ-13-01-16)~~

摘　　要：利用基因工程手段重组表达了弗氏柠檬酸杆菌来源的酪氨酸酚裂解酶(TPL),以丙酮酸和L-丝氨酸为底物全细胞催化合成L-酪氨酸,考察了pH、温度、表面活性剂、金属离子、铵盐种类和氯化铵浓度等因素对L-酪氨酸合成的影响,并比较了两种底物合成L-酪氨酸的转化率。结果表明,TPL的最适反应条件是45℃,pH=8.0,4mmol/L PLP,氯化铵浓度350 mmol/L。1 mmol/L triton-x 100对TPL酶活有促进作用,金属离子对TPL酶活都有明显的抑制作用。0.1 mol/L丙酮酸的转化率约是L-丝氨酸的1.8倍。The tyrosine phenol lyase（TPL） from Citrobacterfreundii was recombinantly expressed in E. coli BL21 （DE3） with the vector pETDuet - 1. The enzymatic properties of the recombinant tyrosine phenol lyase were studied, and several influence factors of enzyme reaction such as pH, temperature, surfactants, metal ions, ammonium salts, and ammonium chloride concentration were investigated. The results indicate that the optimum reaction conditions of TPL were 45 ℃, pH = 8.0,4 mmol/L PLP,350 mmol/L NH4C1,1 mmoL/L triton-x 100. The conversion rate of pyruvate substrate was 1.8 times higher than that of serine substrate.

关 键 词：酪氨酸酚裂解酶 L-酪氨酸 丙酮酸 L-丝氨酸 生物工程 

分 类 号：R977.4[医药卫生—药品]