| 论文题目: | Characterization of a recombinant (+)-gamma-lactamase from Microbacterium hydrocarbonoxydans which provides evidence that two enantiocomplementary gamma-lactamases are in the strain |
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| 作者: | Wang Jianjun Zhu Yaxin, Zhao Guogang, Zhu Junge, Wu Sheng*. |
| 联系作者: | |
| 刊物名称: | Applied Microbiology and Biotechnology |
| 期: | 99 |
| 卷: | 7 |
| 页: | 3069-3080 |
| 年份: | 2015 |
| 影响因子: | 3.848 |
| 论文下载: | http://link.springer.com/article/10.1007%2Fs00253-014-6114-8 |
| 摘要: | A two-step method, i.e., the transfer acyl analysis and then the chiral HPLC analysis, was employed in the screening of the cosmid library of Microbacterium hydrocarbonoxydans genome. Two enantiocomplementary gamma-lactamase clones were found. A 40-kb cosmid showed (-)-gamma-lactamase activity, and the activity was from Mhg which was reported previously according to the results of PCR identifying experiment. The 37-kb (+)-gamma-lactamase cosmid was further constructed into a pUC18 plasmid library and screened by the same two-step method. A plasmid clone harboring a 1.6-kb fragment showed (+)-gamma-lactamase activity. A 555-bp ORF in the 1.6-kb fragment showed high (+)-gamma-lactamase activity when it was expressed under the control of T-7 promoter. The coding protein showed significant homology with bacterial isochorismatase. The (+)-gamma-lactamase was characterized and compared with the (-)-gamma-lactamase Mhg. This is another report that two enantiocomplementary gamma-lactamases are present in the same strain. |
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