| 催化合成谷胱甘肽的基因工程菌的构建及表达 |
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| 引用本文： | 沈继朵,胡晓川,卞筱泓,许激扬.催化合成谷胱甘肽的基因工程菌的构建及表达[J].西北药学杂志,2010,25(4):288-291. |
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| 作者姓名： | 沈继朵 胡晓川 卞筱泓 许激扬 |
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| 作者单位： | 1. 中国药科大学生物化学教研室,江苏,南京,210009
2. 青岛市中心医院乳腺外科,山东,青岛,266042 |
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| 摘 要： | 目的利用基因工程菌合成谷胱甘肽。方法克隆γ-谷氨酰转肽酶,构建重组质粒pET30a-GGT,乳糖诱导融合蛋白的表达,利用pET30a-GGT/E.coliBL21和pET32a-GSH-II/E.coliBL21分两步法合成γ-谷氨酰半胱氨酸与谷胱甘肽。结果工程菌的最佳乳糖诱导条件为:浓度3mmoL.L-1、时间6h、温度28℃。工程菌经最佳条件诱导后,γ-谷氨酰转肽酶的酶活大约是出发菌株E.coliDH5α的21倍,酶活力得到明显提高,谷胱甘肽的产量达到0.524g.L-1。结论成功地为今后利用基因工程菌催化合成谷胱甘肽寻找到一种新的方法 。
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| 关 键 词： | γ-谷氨酰转肽酶 克隆 表达 谷胱甘肽合成 |
Construction of recombinant bacteria for glutathione biosynthesis and expression of recombinant gene |
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| SHEN Jiduo,HU Xiaochuan,BIAN Xiaohong,XU Jiyang.Construction of recombinant bacteria for glutathione biosynthesis and expression of recombinant gene[J].Northwest Pharmaceutical Journal,2010,25(4):288-291. |
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| Authors: | SHEN Jiduo HU Xiaochuan BIAN Xiaohong XU Jiyang |
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| Institution: | ( Department of Biochemistry, China Pharmaceutical University, Nanjing 210009, China ; Breast Surgery, Qingdao Central Hospital, Qingdao , 266042, China ) |
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| Abstract: | Objective To produce glutathione by engineering bacteria. Methods The gene of γ-glutamy/transpeptidase was cloned into vector pET30a. The recombinant protein was induced by lactose. E. coli BL21（pET30a-GGT） together with E. coli BL21（pET32a- GSH-II） used to produce GSH by two-step faction. Results The optimial lactose induction conditions of ）γ-glutamyl transpeptidase were as follows：concentration 3 mmol.L^-1 , time 6 h, and temperature 28℃. After lactose induction, the activity of γ-glutamyl transpeptidase of the engineered strain was 21 times higher than that of the starting bacterium. The yield of glutathione was 0. 524 g.L^-1. Conelusion These results found a new rneathod for glutathione synthesis by engineering bacteria. |
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| Keywords: | γ-glutamyl transpeptidase Clone Expression glutathione synthesis |
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