| 一株谷氨酰胺合成酶高产菌株的酶学分析及其基因的克隆分析 |
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| 引用本文： | 肖钢,邱鑫,丁邦琴.一株谷氨酰胺合成酶高产菌株的酶学分析及其基因的克隆分析[J].湖北大学学报(自然科学版),2010,32(3):326-329. |
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| 作者姓名： | 肖钢 邱鑫 丁邦琴 |
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| 作者单位： | 肖钢(湖南农业大学油料作物研究所,湖南,长沙,410128);邱鑫,丁邦琴(南通职业大学化工系,江苏,南通,226007) |
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| 基金项目： | 南通市科技局应用研究计划 |
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| 摘 要： | 对谷氨酰胺合成酶菌球形节杆菌(Arthrobacter globiformis)突变菌株Y3进行分析.其谷氨酰胺合成酶摇瓶发酵单位是原始菌株的2.81倍;最适反应温度为40℃;最适pH为6.5.以突变菌株和原始菌株基因组为模板,通过同源克隆策略,成功地获得了球形节杆菌谷氨酰胺合成酶基因gln片段.通过序列比对,突变菌株gln基因在第409 bp处产生T→C的碱基的突变,该处突变使酪氨酸(Tyr409)被组氨酸(His409)取代,该突变解决了谷氨酰胺合成酶腺苷酰基化问题,保证了其在高浓度铵盐条件下保持酶活性.
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| 关 键 词： | 酶菌球形节杆菌（Arthrobacter globiformis） 紫外诱变 谷氨酰胺合成酶 酶活 |
Analysis of a high glutamine synthetase producing Arthrobacter globiformis strain |
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| XIAO Gang,QIU Xin,DING Bangqin.Analysis of a high glutamine synthetase producing Arthrobacter globiformis strain[J].Journal of Hubei University(Natural Science Edition),2010,32(3):326-329. |
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| Authors: | XIAO Gang QIU Xin DING Bangqin |
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| Institution: | 1.The Oil Crops Institute of Hunan Agricultural University,Changsha 410128,China;2.Department of Chemical Engineering,Nantong Vocational College,Nantong 226007,China) |
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| Abstract: | In this study,a mutant strain Y3 with high glutamine synthetase productivity was investigated,its activity was 2.81-fold of the original one.Examined the nature of glutamine synthetase,the results showed that the optimal reaction temperature and pH was 40 ℃ and 6.5,respectively.According to the homology,we designed a pair of PCR primers to clone the gln gene from UV mutant and original strain of Arthrobacter globiformis,respectively.Aligment of the glns revealed that a transversion of T→C at the position 409 bp,and this made Tyr409 was replaced by His409.The mutant of T→C can solve deactivation of glutamine synthetase（GS） resulting from adenylylation in high NH＋4 concentrations. |
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| Keywords: | globiformis) |
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