文章摘要
潘鑫茹,焦庆才,刘均忠,张宏娟.共表达PPK和GMAS全细胞催化合成L-茶氨酸[J].精细化工,2019,36(9):
共表达PPK和GMAS全细胞催化合成L-茶氨酸
Synthesis of L-theanine by Whole Cell Catalysis with co-expression of PPK and GMAS
投稿时间:2019-01-02  修订日期:2019-03-08
DOI:
中文关键词: 共表达  多聚磷酸盐激酶  γ-谷氨酰甲胺合成酶  L-茶氨酸  ATP再生
英文关键词: co-expression  polyphosphate kinase  γ-glutamylmethylamide synthetase  L-theanine  ATP regeneration
基金项目:国家自然科学基金(21302100)
作者单位E-mail
潘鑫茹 南京大学生命科学学院 m15774605689@163.com 
焦庆才 南京大学 jiaoqc@163.com 
刘均忠 南京大学生命科学学院  
张宏娟 南京医科大学 药学院  
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中文摘要:
      利用pETDuet-1质粒在大肠杆菌BL21(DE3)中重组表达了多聚磷酸盐激酶(PPK)和 γ-谷氨酰甲胺合成酶(GMAS),并以共表达PPK和GMAS的重组菌全细胞催化合成L-茶氨酸,优化了反应参数。SDS-PAGE结果表明PPK和GMAS共表达成功;最佳全细胞催化反应条件为:35 oC,pH=7.0,谷氨酸钠300 mmol/L,乙胺盐酸盐420 mmol/L,六偏磷酸钠100 mmol/L,添加2 mmol/L 起始量ATP,在100 mL反应体系中转化24 h L-茶氨酸浓度达到199 mmol/L,转化率达到66.34%。
英文摘要:
      Polyphosphate kinase (PPK) and γ-glutamylmethylamide synthetase (GMAS) were recombinant expressed using pETDuet-1 plasmid in Escherichia coli BL21(DE3), and L-theanine was synthesized by whole cell catalysis with co-expression of PPK and GMAS. The parameters of reaction were investigated and optimized. The SDS-PAGE analysis showed that PPK and GMAS were successfully co-expressed in the recombinant strains. The optimal conditions for whole cell biocatalytic reaction were as follows: 35 oC, pH=7.0, 300 mmol/L monosodium glutamate, 420 mmol/L ethylamine hydrochloride, 100 mmol/L polyP, 2 mmol/L ATP. After 24 h in a 100 mL reaction system, under the optimal conditions the L-theanine yield can reach 199 mmol/L, and the conversion rate reaches 66.34%.
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