[1]王期,抗晶晶,忻寅强,等.生物转化法重组谷氨酸脱羧酶合成γ-氨基丁酸[J].南京师大学报(自然科学版),2010,33(03):85-90.
　Wang Qi,Kang Jingjing,Xing Yinqiang,et al.Production of γ-aminobutyric Acid(γ-GABA) by Bioconversion With Recombinant Glutamate Decarboxylase[J].Journal of Nanjing Normal University(Natural Science Edition),2010,33(03):85-90.
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生物转化法重组谷氨酸脱羧酶合成γ-氨基丁酸()
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《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第33卷
期数:
2010年03期
页码:
85-90
栏目:
生命科学
出版日期:
2010-09-20

文章信息/Info

Title:
Production of γ-aminobutyric Acid(γ-GABA) by Bioconversion With Recombinant Glutamate Decarboxylase
作者:
王期;抗晶晶;忻寅强;李桂兰;殷志敏;
南京师范大学生命科学学院, 江苏南京210046
Author(s):
Wang Qi; Kang Jingjing; Xing Yinqiang; Li Guilan; Yin Zhimin
School of Life Science,Nanjing Normal University,Nanjing 210046,China
关键词:
谷氨酸脱氢酶( GAD) ;  工程菌;  酶法合成;  C-氨基丁酸( GABA )
Keywords:
G lutam ate decarboxy lase;  eng ineered stra in;  enzym atic synthesis; γ-GABA
分类号:
Q78
摘要:
成功构建了一个高效表达大肠杆菌谷氨酸脱羧酶GAD来源的重组质粒pET28a-gadA,并转化E.coli BL21(DE3),工程菌株经0.4 mmol/L IPTG或1 g/L的乳糖,37℃诱导表达8 h,粗酶液的酶活达到12 μ/mL,大约是出发菌株E.coliK-12的60倍.工程菌1.15 U粗酶液以31 g/L L-谷氨酸钠为底物,37℃、pH4.0条件下反应4 h,γ-氨基丁酸的生成量达到19.57 g/L,L-谷氨酸钠的转化率为93%,从而为γ-氨基丁酸的生产提供了很好的前景
Abstract:
A recomb inant p lasm id nam ed pET28a-gadA which could high ly produce g lutama te decarboxy lase was constructed. The recomb inant p lasm id w as transfo rm ed to E. coli BL21 ( DE3 ). Gutam ate deca rboxy lase was produced at 37℃ , which w as incubated w ith 014 mm o l/L IPTG or 1 g /L lactose fo r 8 h. The ac tiv ity o f 1 mL crude ex tract o f recomb inant stra in was usually 12 U /mL, w hich was about 60 tim es higher than the activ ity o fE. co li K- 12. In 1mL reaction system, when 31 g /L L-G luN aw as used, 1115U crude ex tract, pH 41 0, and incubation a t 37℃ fo r 4 h w ere the opt-i mum cond ition. The y ie ld ofC-GABA w as 191 57 g /L. F inally, the rate o f conversion from L-G luN a to theC-GABA w as up to 93%, wh ich ev entually can be prov ided a good prospect fo r the production o fC-am inobutyr ic ac id.

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备注/Memo

备注/Memo:
通讯联系人: 殷志敏, 博士, 教授, 研究方向: 生物化学及细胞生物学. E-m ail:yinzhim in@njnu. edu. cn
更新日期/Last Update: 2013-04-08