| 重组L-苏氨酸醛缩酶催化合成L-4-硝基苯基丝氨酸 |
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| 引用本文： | 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). |
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| 作者姓名： | 余进海 吴婷 刘均忠 张宏娟 焦庆才 |
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| 作者单位： | 南京大学,南京大学,南京大学,南京医科大学,南京大学 |
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| 基金项目： | 国家自然科学基金青年科学基金项目 |
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| 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%。
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| 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 |
| 收稿时间： | 2018/1/19 0:00:00 |
| 修稿时间： | 2018/5/2 0:00:00 |
Enzymatic Synthesis of L-4-Nitrophenylserine by Recombinant L-Threonine Aldolase |
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| yujinhai,wuting,liujunzhong,zhanghongjuan and jiaoqingcai.Enzymatic Synthesis of L-4-Nitrophenylserine by Recombinant L-Threonine Aldolase[J].Fine Chemicals,2018,35(12). |
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| Authors: | yujinhai wuting liujunzhong zhanghongjuan and jiaoqingcai |
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| Affiliation: | Nanjing university,Nanjing university,Nanjing university,Nanjing Medical University,Nanjing university |
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| Abstract: | 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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| Keywords: | L-threonine aldolase 4-nitrobenzaldehyde L-4-nitrophenylserine active carbon adsorption column |
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