| 大肠杆菌精氨酰－tRNA合成酶高表达条件的优化及酶的纯化 |
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| 引用本文： | 黄意巍,王恩多,王应睐. 大肠杆菌精氨酰－tRNA合成酶高表达条件的优化及酶的纯化[J]. Acta biochimica et biophysica Sinica, 1995, 0(3) |
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| 作者姓名： | 黄意巍 王恩多 王应睐 |
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| 作者单位： | 中国科学院上海生物化学研究所分子生物学国家重点实验室 |
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| 摘 要： | 控制培养基中氨苄青霉素的用量、ｐＨ和培养时间，从含Ｅ．ｃｏｌｉａｒｇｙ变种ａｒｇｒ３８１ＫＡ的Ｅ．ｃｏｌｉＴＧ１转化子中，得到了Ｅ．ｃｏｌｉＡｒｇＲＳ变种ＡｒｇＲＳ３８１ＫＡ的高表达。从２升培养液中得到１５克湿菌体，粗抽液中ＡｒｇＲＳ３８１ＫＡ的比活为５０３单位／毫克。经过两次ＤＥＡＥＳｅｐｈａｃｅｌ层析，在４天时间内，可得到７８毫克电泳一条带的纯酶活力回收达８０％。该方法可以作为从含ａｒｇｓ的Ｅ．ｃｏｌｉＴＧ１转化子中提纯Ｅ．ｃｏｌｉＡｒｓＲＳ的通用方法。
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| 关 键 词： | 大肠杆菌，精氨酸－tRNA合成酶，纯化，高表达 |
Overproduction and Purification of Arginyl-tRNA Synthetese from E. coli |
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| HUANG Yi-Wei, WANG En-Duo and WANG Ying-Lai. Overproduction and Purification of Arginyl-tRNA Synthetese from E. coli[J]. Acta biochimica et biophysica Sinica, 1995, 0(3) |
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| Authors: | HUANG Yi-Wei WANG En-Duo WANG Ying-Lai |
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| Abstract: | The mutant of Arginyl-tRNA synthetase, ArgRS381KA, was overproduced from the transformat harbouring the recombinant plasmid which contains args381KA, the mutant gene of this enzyme, by controlling the amount of ampicilin in the culture, the pH, and the time of incubation. Fifteen gram of wet cells had been obtained from a 2 liter culture, the specific activity of ArgRS381KA in the crude extract was 503 U/mg. By two steps of DEAE-Sephacel column chromatography, 78 mg of purified ArgRS381KA had been obtained in 4 days. The recovery of activity of this enzyme was 80 %. This method may be used as a general method for the purification of Arginyl-tRNA synthetase (ArgRS) from E. coli transformat contoining the gene of ArgRS(args). |
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| Keywords: | E. coli Arginyl-tRNA synthetase Purification Overproducotion |
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