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International Journal of Medicinal Mushrooms
IF: 1.423 5-Year IF: 1.525 SJR: 0.433 SNIP: 0.661 CiteScore™: 1.38

ISSN Print: 1521-9437
ISSN Online: 1940-4344

International Journal of Medicinal Mushrooms

DOI: 10.1615/IntJMedMushrooms.v18.i6.90
pages 547-554

Purification and Characterization of a Novel Serine Protease from the Fruiting Bodies of a Rare Edible Medicinal Mushroom, Lyophyllum shimeji (Agaricomycetes)

Xueran Geng
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing, China
Rigen Te
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing, China
Guoting Tian
Institute of Biotechnology and Germplasm Resource, Yunnan Academy of Agricultural Science, Kunming, China
Yongchang Zhao
Institute of Biotechnology and Germplasm Resource, Yunnan Academy of Agricultural Science, Kunming, China
Liyan Zhao
College of Food Science and Technology, Nanjing Agricultural University, Weigang, Nanjing, China
Hexiang Wang
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China
Tzi Bun Ng
School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China

ABSTRACT

In this study, a novel protease with a molecular mass of 30 kDa was purified from Lyophyllum shimeji using a purification procedure that involved anion exchange chromatography on a Q-Sepharose column, cation chromatography on an SP-Sepharose column, and gel filtration on a Superdex 75 column. The protease was purified 57-fold, and its specific activity was 6.67 U/mg. Its inner amino acid sequence, determined by liquid chromatography-tandem mass spectrometry, contains AASIIAVLVLSDK, which has 93% identity with the sequence of Hypsizygus marmoreus serine protease. The optimal reaction temperature and pH for L. shimeji protease were 50°C and 10.0, respectively. It is an alkaline protease and has higher activity when the pH lies between 6.8 and 10.0. The Km and Vmax at 50°C and pH 9.0 were 1.32 mg/mL and 454.55 μg/mL/min, respectively. The activity of L. shimeji protease was significantly suppressed by Cd2+, Hg2+, Cu2+, and Fe3+ ions, as well as by phenylmethylsufonyl fluoride; therefore, it is a serine protease.


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