| 论文题目: | New strategy of site-directed mutagenesis identifies new sites to improve Streptomyces clavuligerus deacetoxycephalosporin C synthase activity toward penicillin G |
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| 作者: | Ji, JJ; Tian, XY; Fan, KQ; Yang, KQ |
| 联系作者: | Fan, KQ |
| 刊物名称: | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY |
| 期: | 6 |
| 卷: | 93 |
| 页: | 2395-2401 |
| 年份: | 2012 |
| 影响因子: | 3.425 |
| 论文下载: | 下载地址 |
| 摘要: | Based on multiple sequence alignment of different deacetoxycephalosporin C synthase (DAOCSs) and the crystal structure of Streptomyces clavuligerus DAOCS, 2-oxoglutarate, and penicillin G triple complex, ten residues (Y184, V245, S261, C37, T42, V51, S59, A61, Q126, and T213) not directly involved in substrate recognition were selected as mutational targets. Twenty one mutants were generated and characterized, and five (Q126M, T213V, S261M, S261A, and Y184A) showed improved activity toward penicillin G, with 1.45- to 4.50-fold increment in the k (cat)/K (m). Q126, T213, and S261 are identified for the first time, as sites with significant effect on enzyme activity. |
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