Purification and characterization of a novel metalloprotease from fruiting bodies of Oudemansiella radicata

  • Xueran Geng College of Food Science and Engineering, Shanxi Agricultural University
  • Rigen Te State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University
  • Guoting Tian Institute of Biotechnology and Germplasmic Resource, Yunnan Academy of Agricultural Science
  • Yongchang Zhao Institute of Biotechnology and Germplasmic Resource, Yunnan Academy of Agricultural Science
  • Liyan Zhao College of Food Science and Technology, Nanjing Agricultural University, Weigang
  • Hexiang Wang State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University
  • Tzi Bun Ng School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong
DOI: https://doi.org/10.18388/abp.2016_1431

Abstract

In this study, a 39-kDa metalloprotease was purified from a rare health-promoting edible mushroom Oudemansiella radicata using a purification protocol which entailed anion exchange chromatography on DEAE-cellulose and Q-Sepharose columns and gel filtration by fast protein liquid chromatography on Superdex 75 column. Some peptide sequences were determined by LC-MS/MS analysis and one of the sequences, DAWIQADVNR, manifested 90% identity to Coprinopsis cinerea metalloprotease. The optimal reaction pH and temperature for Oudemansiella radicata protease were pH 7.0 and 50℃, respectively. The protease was purified 79-fold and demonstrated a specific protease activity of 2.42 U/mg. The Km of the purified protease for casein was 0.65 mg/mL at pH 7.0 and 50 ℃.The activity of the protease was inhibited by Cd2+、Hg2+、Cu2+、Pb2+ and Fe3+ ions, and was enhanced by K+ ions. The activity of Oudemansiella radicata protease was strongly suppressed by EDTA, indicating that it is a metalloprotease.

References

Choi, B.S., Sapkota, K., Choi, J.H., Shin, C.H., Kim, S., and Kim, S.J. (2013) Herinase: a novel bi-functional fibrinolytic protease from the monkey head mushroom, Hericium erinaceum. Appl Biochem Biotechnol 170:609-622.http://dx.doi.org/10.1007/s12010-013-0206-2

Published
2017-09-08
Issue
Vol. 64 No. 3 (2017)
Section
Articles

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