Abstract
In this study, a novel metallo-β-lactamases fold hydrolase PH-1 was identified from Pelagibacterium halotolerans B2T. This novel member of the family Hyphomicrobiaceae was isolated from the East China Sea. In silico analysis demonstrated that PH-1 and its relative homologues cluster in a unique branch and constitute a new subgroup among MBLs. PH-1 was cloned and overexpressed in Escherichia coli BL21 in a soluble form. SDS-PAGE, MALDI-TOF/TOF–MS, and size-exclusion chromatography analysis demonstrated that the PH-1 was a monomer with molecular weight of about 29 kDa. Substrate specificity study showed PH-1 preferred penicillin type β-lactams and exhibited maximum activity toward penicillin-G. Additionally, our experiments also revealed that PH-1 was a halotolerant enzyme since it is active under 4 M NaCl. The enzyme activity of PH-1 was negatively affected by 1 mM Mn2+ and EDTA. These observations lay a foundation for further study of MBLs from marine bacterium.
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Acknowledgments
We thank Dr. Sunjay Mannan and Huihui Dong for manuscript polishing. This project was supported by the National Natural Science Foundation of China (Grant Nos. 81301461 and 41406140) and Zhejiang Provincial Natural Science Foundation of China (Grant No. LQ13H190002).
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Communicated by S. Albers.
B. Zheng and X. Jiang contributed equally to the study.
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Zheng, B., Jiang, X., Xu, Z. et al. Characterization of a novel metallo-β-lactamases fold hydrolase from Pelagibacterium halotolerans, a marine halotolerant bacterium isolated from East China Sea. Extremophiles 20, 37–44 (2016). https://doi.org/10.1007/s00792-015-0795-5
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DOI: https://doi.org/10.1007/s00792-015-0795-5