L-苏氨酸醛缩酶催化合成L-4-硝基苯基丝氨酸  

Synthesis of 4-Nitrophenylserine by Whole Cell with Recombinant L-Threonine Aldolase

作　　者：余进海 吴婷 刘均忠[1] 张宏娟[2] 焦庆才[1] YU Jin-hai;WU Ting;LIU Jun-zhong;ZHANG Hong-juan;JIAO Qing-cai(State Key Laboratory of Pharmaceutical Biotechnology,School of Life Sciences,Nanjing University,Nanjing 210093,Jiangsu,China;School of Pharmacy,Nanjing Medical University,Nanjing 210029,Jiangsu,China)

机构地区：[1]医药生物技术国家重点实验室南京大学生命科学学院,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》2018年第12期2039-2044,2064,共7页Fine Chemicals

基　　金：国家自然科学基金青年科学基金项目(21302100)~~

摘　　要：利用pGEX-KG载体在大肠杆菌BL21(DE3)中重组表达L-苏氨酸醛缩酶,以对硝基苯甲醛、甘氨酸为底物,采用酶法合成了L-4-硝基苯基丝氨酸,优化了反应条件,得到的最佳条件为:反应温度45℃,pH=8.0,甘氨酸浓度500mmol/L,对硝基苯甲醛浓度100mmol/L,每升转化液含1.6g全细胞,反应24h后,L-4-硝基苯基丝氨酸质量浓度达到9.72g/L,对硝基苯甲醛转化率为43%。放大至500mL后转化液经过活性炭分离纯化得到产物3.92g,总收率为35%。The recombinant pGEX-KG plasmid was constructed in Escherichia coli BL21(DE3)and expressed with L-threonine aldolase.Then,the obtained L-threonine aldolase was used as a catalyst for the synthesis of L-4-nitrophenylserine from 4-nitrobenzaldehyde and glycine.The reaction conditions were optimized.The results showed that the optimal conditions were as follows:reaction temperature was 45℃,pH value was 8.0,the concentration of glycine was 500 mmol/L,the concentration of 4-nitrobenzaldehyde was 100 mmol/L,the mass concentration ofL-threonine aldolase whole cell was 1.6 g/L,and reaction time was 24 h. Under these conditions,the mass concentration of L-4-nitrophenylserine reached 9.72g/L and the conversion of 4-nitrobenzaldehyde was 43%.After amplification experiment (500mL),the conversion solution was isolated and further purified by active carbon,3.96g L-4-nitrophenylserine was obtained with a total yield of 35%.

关 键 词：L-苏氨酸醛缩酶 对硝基苯甲醛 L-4-硝基苯基丝氨酸 活性炭吸附柱 生物工程 

分 类 号：Q55[生物学—生物化学]