4-丁内酰胺水解酶法制备γ-氨基丁酸  

Preparation of Gamma-Amino Butyric Acid Using γ-Lactamase

作　　者：蒋光玉 刘洋 汪艳 

机构地区：[1]安徽华恒生物工程有限公司,合肥231131

出　　处：《氨基酸和生物资源》2011年第1期43-48,共6页Amino Acids & Biotic Resources

基　　金：合肥市科技攻关项目(No.20081024)资助

摘　　要：以吡咯烷酮为唯一碳源,从采集的土样中分离、筛选得到具有4-丁内酰胺水解酶活性的菌株。采用响应面分析法对该菌株的产酶培养基组分进行了优化研究并对酶促转化反应条件也进行了研究。结果表明:编号为HHSW-16的菌株水解活性最高。优化后的培养基组成为:葡萄糖11.50 g.L-1,牛肉膏6.35 g.L-1,酵母粉5.58 g.L-1,Na2HPO4.12H2O 5.0g.L-1,KH2PO4 1.5 g.L-1,MgSO4.7H2O 1.0 g.L-1,CaCl2.2H2O 10 mg.L-1,FeSO4.7H20 8 mg.L-1,pH7.0。转化反应的最佳条件为:温度40℃,pH值7.0。此条件下,10 g.L-1底物加入1%湿菌体,酶促反应6 h转化率达到99%以上。Strains capable of producing γ-lactamase were screened from soil samples collected in several districts,and particularly 2-pyrrolidone was used as sole carbon source.Fermentation medium was optimized using response surface analysis method and enzymatic reactions for gamma-amino butyric acid（GABA） production were optimized.Results showed that： the strain named HHSW-16 had the best hydrolysis activity.The best fermentation medium was glucose 11.50g·L-1,beef extract 6.35g·L-1,yeast extract 5.58g·L-1,Na 2HPO 4·12H 2O 5.0g·L-1,KH 2PO 4 1.5g·L-1,MgSO 4·7H 2O 1.0g·L-1,CaCl 2·2H 2O 10mg·L-1,FeSO 4·7H 2O 8mg·L-1,pH 7.0.The optimum enzymatic reaction conditions were as follows：temperature 40℃,pH 7.0.Under these conditions,10g·L-1 substrate and 1% wet biomass,giving the reaction 6 hours,transformation ratio yield was up to 99.0%.

关 键 词：Γ-氨基丁酸 4-丁内酰胺水解酶 2-吡咯烷酮 

分 类 号：Q5[生物学—生物化学]