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Characterization of a recombinant (+)-γ-lactamase from Microbacterium hydrocarbonoxydans which provides evidence that two enantiocomplementary γ-lactamases are in the strain

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Abstract

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 γ-lactamase clones were found. A 40-kb cosmid showed (−)-γ-lactamase activity, and the activity was from Mhg which was reported previously according to the results of PCR identifying experiment. The 37-kb (+)-γ-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 (+)-γ-lactamase activity. A 555-bp ORF in the 1.6-kb fragment showed high (+)-γ-lactamase activity when it was expressed under the control of T7 promoter. The coding protein showed significant homology with bacterial isochorismatase. The (+)-γ-lactamase was characterized and compared with the (−)-γ-lactamase Mhg. This is another report that two enantiocomplementary γ-lactamases are present in the same strain.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China no. 31070718.

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Authors and Affiliations

  1. State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 Beichen West Road, Chaoyang, Beijing, 100101, People’s Republic of China

    Jianjun Wang, Yaxin Zhu, Junge Zhu & Sheng Wu

  2. Department of Internal Medicine, College of Medicine, University of Kentucky, Lexington, KY, 40536, USA

    Guogang Zhao

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  1. Jianjun Wang
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  2. Yaxin Zhu
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Correspondence to Sheng Wu.

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Wang, J., Zhu, Y., Zhao, G. et al. Characterization of a recombinant (+)-γ-lactamase from Microbacterium hydrocarbonoxydans which provides evidence that two enantiocomplementary γ-lactamases are in the strain. Appl Microbiol Biotechnol 99, 3069–3080 (2015). https://doi.org/10.1007/s00253-014-6114-8

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