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

• Authors: Xueran Geng , Rigen Te , Guoting Tian , Yongchang Zhao , Liyan Zhao , Hexiang Wang , Tzi Ng
• Languages of publication: EN

Abstracts

EN

In this study, a 39-kDa metalloprotease was purified from a rare edible mushroom with health-promoting activities, 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 a Superdex 75 column. Some peptide sequences were obtained 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°C, 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 the casein substrate was 0.65 mg/mL at pH 7.0 and 50°C. The activity of the protease was inhibited by Cd2+, Hg2+, Cu2+, Pb2+ and Fe3+ ions, but was enhanced by K+, Mn2+ and Fe2+ ions. The marked suppression of the protease activity by EDTA indicates that the protease is a metalloprotease.

Keywords

EN

edible mushroom; Oudemansiella radicata; protease; purification

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2017
• Volume: 64
• Issue: 3
• Pages: 477-483

Dates

published

2017

received

2016-09-26

revised

2017-03-10

accepted

2017-05-08

(unknown)

2017-09-08

Contributors

author

Xueran Geng

• College of Food Science and Engineering, Shanxi Agricultural University, Taigu, Shanxi 030801, China
• State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China

author

Rigen Te

• State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China

author

Guoting Tian

• Institute of Biotechnology and Germplasmic Resource, Yunnan Academy of Agricultural Science, Kunming 650223, China

author

Yongchang Zhao

• Institute of Biotechnology and Germplasmic Resource, Yunnan Academy of Agricultural Science, Kunming 650223, China

author

Liyan Zhao

• College of Food Science and Technology, Nanjing Agricultural University, Weigang, Nanjing 210095, China

author

Hexiang Wang

• State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China

author

Tzi Ng

• School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China

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Identifiers

DOI

10.18388/abp.2016_1431