文章摘要
余进海,吴婷,刘均忠,张宏娟,焦庆才.重组L-苏氨酸醛缩酶催化合成L-4-硝基苯基丝氨酸[J].精细化工,2018,35(12):
重组L-苏氨酸醛缩酶催化合成L-4-硝基苯基丝氨酸
Enzymatic Synthesis of L-4-Nitrophenylserine by Recombinant L-Threonine Aldolase
投稿时间:2018-01-19  修订日期:2018-05-02
DOI:
中文关键词: L-苏氨酸醛缩酶  4-硝基苯甲醛  L-4-硝基苯基丝氨酸  活性炭吸附柱
英文关键词: L-threonine aldolase, 4-nitrobenzaldehyde, L-4-nitrophenylserine, active carbon adsorption column
基金项目:国家自然科学基金青年科学基金项目
作者单位E-mail
余进海 南京大学 yujinhai325@outlook.com 
吴婷 南京大学  
刘均忠 南京大学  
张宏娟 南京医科大学  
焦庆才 南京大学 jiaoqc@163.com 
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中文摘要:
      利用pGEX-KG载体在大肠杆菌BL21(DE3)中重组表达了L-苏氨酸醛缩酶,以4-硝基苯甲醛、甘氨酸为底物酶法合成了L-4-硝基苯基丝氨酸,考察了反应温度、pH、底物摩尔比和甘氨酸浓度对酶活的影响。最佳转化条件为:反应温度45℃,pH=8.0,甘氨酸与4-硝基苯甲醛底物摩尔比5:1,底物甘氨酸最适浓度为500 mmol/L;0.1 g湿细胞菌体在10 mL反应体系中在最佳反应条件下反应24 h,底物4-硝基苯甲醛转化率为43%,产物L-4-硝基苯基丝氨酸达到9.72g/L,总收率为35%。
英文摘要:
      Enzymatic synthesis of L-4-nitrophenylserine from glycine and 4-nitrobenzaldehyde catalyzed by recombinant L-threonine aldolase in Escherichia coli BL21(DE3) with pGEX-KG plasmid was studied. Some factors influencing enzymatic reaction, such as temperature, pH, molar ratio of glycine to 4-nitrobenzaldehyde and substrate concentration were investigated. The optimal temperature and pH value were 45oC and 8.0. The optimal molar ratio of glycine to 4-nitrobenzaldehyde was 5:1, The optimal glycine concentration was 500 mmol/L. Under the optimal conditions, the bioconversion rate of 4-nitrobenzaldehyde reached 43% after 24 h. The concentration of L-4-nitrophenylserine reached 9.72 g/L, with yield of 35%.
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