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Peptidyl‐Lys metalloendopeptidase (Lys‐N) purified from dry fruit of Grifola frondosa demonstrates “mirror”digestion property with lysyl endopeptidase (Lys‐C)
Author(s) -
Zhao Mingzhi,
Hao Bingbing,
Li Honghao,
Cai Man,
Xie Jingjing,
Liu Hongyu,
Tan Minjia,
Zhai Linhui,
Yu Qun
Publication year - 2020
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.8573
Subject(s) - chemistry , grifola frondosa , proteomics , chromatography , endopeptidase , lysine , biochemistry , tandem mass spectrometry , enzyme , mass spectrometry , amino acid , polysaccharide , gene
Rationale Lys‐N, also known as lysine‐specific metalloendopeptidase, functions as the “sister” enzyme of lysyl endopeptidase (Lys‐C) in proteomic research. Its digestion specificity at the N‐terminal lysine residue makes it a very useful tool in proteomics analysis, especially in mass spectrometry (MS)‐based de novo sequencing of proteins. Methods Here we present a complete production process of highly purified Lys‐N from dry fruit of Grifola frondosa (maitake mushroom). The purification process includes one step of microfiltration plus one step of UF/DF (ultrafiltration used in tandem with a diafiltration method) recovery and four steps of chromatographic purification. Results The overall yield of the process was approximately 6.7 mg Lys‐N protein/kg dry fruit of G. frondosa . The assay data demonstrated that the purified Lys‐N exhibited high enzymatic activity and specificity. Conclusions The novel production process provides for the first time the extraction of Lys‐N from dry fruit of G. frondosa . The process is also stable and scalable, and provides an economic way of producing the enzyme in large quantities for MS‐based proteomics and other biological studies.