Peptidyl-Lys metalloendopeptidase (Lys-N) purified from dry fruit of Grifola frondosa demonstrates "mirror"digestion property with lysyl endopeptidase (Lys-C)
文献类型：期刊论文
| 作者 | Zhao, Mingzhi3,4; Hao, Bingbing5,6; Li, Honghao1; Cai, Man3; Xie, Jingjing3; Liu, Hongyu3; Tan, Minjia5,6 ; Zhai, Linhui2,5; Yu, Qun4
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| 刊名 | RAPID COMMUNICATIONS IN MASS SPECTROMETRY
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| 出版日期 | 2020-01-30 |
| 卷号 | 34期号:2页码:11 |
| ISSN号 | 0951-4198 |
| DOI | 10.1002/rcm.8573 |
| 通讯作者 | Zhai, Linhui(zhailinhui@simm.ac.cn) ; Yu, Qun(yuqun1970@outlook.com) |
| 英文摘要 | 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. |
| WOS关键词 | ARMILLARIA-MELLEA ; HETEROLOGOUS EXPRESSION ; PROTEIN IDENTIFICATION ; MASS-SPECTROMETRY ; PURIFICATION ; METALLOPEPTIDASE ; ACTIVATION ; MUSHROOM ; TRYPSIN ; CLONING |
| 资助项目 | China Postdoctoral Science Foundation[2018 M633742] ; National Natural Science Foundation of China[31670066] ; National Natural Science Foundation of China[31700724] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Chemistry ; Spectroscopy |
| 语种 | 英语 |
| WOS记录号 | WOS:000506630600002 |
| 出版者 | WILEY |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/282377]  |
| 专题 | 新药研究国家重点实验室 |
| 通讯作者 | Zhai, Linhui; Yu, Qun |
| 作者单位 | 1.Sartorius Stedim Shanghai Trading Co Ltd, Shanghai 201203, Peoples R China 2.Nanjing Univ Chinese Med, Nanjing 210023, Peoples R China 3.Prosit Sole Biotechnol Co Ltd, Beijing 100085, Peoples R China 4.Beijing Inst Transfus Med, 27 Taiping Rd, Beijing 100850, Peoples R China 5.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, Chem Prote Ctr, Shanghai 201203, Peoples R China 6.Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhao, Mingzhi,Hao, Bingbing,Li, Honghao,et al. Peptidyl-Lys metalloendopeptidase (Lys-N) purified from dry fruit of Grifola frondosa demonstrates "mirror"digestion property with lysyl endopeptidase (Lys-C)[J]. RAPID COMMUNICATIONS IN MASS SPECTROMETRY,2020,34(2):11. |
| APA | Zhao, Mingzhi.,Hao, Bingbing.,Li, Honghao.,Cai, Man.,Xie, Jingjing.,...&Yu, Qun.(2020).Peptidyl-Lys metalloendopeptidase (Lys-N) purified from dry fruit of Grifola frondosa demonstrates "mirror"digestion property with lysyl endopeptidase (Lys-C).RAPID COMMUNICATIONS IN MASS SPECTROMETRY,34(2),11. |
| MLA | Zhao, Mingzhi,et al."Peptidyl-Lys metalloendopeptidase (Lys-N) purified from dry fruit of Grifola frondosa demonstrates "mirror"digestion property with lysyl endopeptidase (Lys-C)".RAPID COMMUNICATIONS IN MASS SPECTROMETRY 34.2(2020):11. |
入库方式： OAI收割
来源：上海药物研究所
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