Abstract
One clone (ACPGA001) exhibiting penicillin G acylase (PGA) activity was screened from a metagenomic library by using a medium containing penicillin G. A novel PGA gene from the inserted fragment of ACPGA001 was obtained by sequencing. The amino acid sequence of ACPGA001 PGA exhibited <33 % similarity to PGAs retrieved from GenBank. This gene was expressed in Escherichia coli M15 and the recombinant protein was purified and characterized. The ACPGA001 PGA exhibited a maximum activity at 60 °C and showed high activity at pH 4–10 with an optimum pH of 8.0. This enzyme was stable at 40 °C for 70 min with a half-life of 60 min at 55 °C. These beneficial characteristics of ACPGA001 PGA provide some advantages for the potential application of ACPGA001 PGA in industry.
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Acknowledgments
This work was financially supported by Hi-Tech Research and Development Program of China (863 program of China; 2012AA092103), China Ocean Mineral Resources R&D Association (DY125-15-T-06) and Marine Scientific Research Special Foundation for Public Sector Program (201005032).
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Qian Zhang and Hui Xu contributed equally to this study.
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Zhang, Q., Xu, H., Zhao, J. et al. Expression and characterization of a thermostable penicillin G acylase from an environmental metagenomic library. Biotechnol Lett 36, 617–625 (2014). https://doi.org/10.1007/s10529-013-1403-3
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DOI: https://doi.org/10.1007/s10529-013-1403-3