食用级真菌嗜sitophila神经孢子菌纤维蛋白溶解酶的纯化及特性研究

Q2 Chemical Engineering
Xiao lan Liu, Narasimha kumar Kopparapu, Hong chen Zheng, Priti Katrolia, Yong ping Deng, Xi qun Zheng
{"title":"食用级真菌嗜sitophila神经孢子菌纤维蛋白溶解酶的纯化及特性研究","authors":"Xiao lan Liu,&nbsp;Narasimha kumar Kopparapu,&nbsp;Hong chen Zheng,&nbsp;Priti Katrolia,&nbsp;Yong ping Deng,&nbsp;Xi qun Zheng","doi":"10.1016/j.molcatb.2016.10.006","DOIUrl":null,"url":null,"abstract":"<div><p>A fibrinolytic protease was purified from the culture supernatant of a GRAS fungus, <em>Neurospora sitophila</em>. The enzyme displayed a molecular mass of 34<!--> <!-->kDa, as estimated by SDS-PAGE and gel filtration chromatography. The isoelectric point (pI) of the enzyme was 9.3<!--> <!-->±<!--> <!-->0.2 as determined by iso-electric focusing (IEF). It was maximally active at pH 7.6 and 41<!--> <!-->°C and displayed remarkable stability in a wide pH range (4–11) and up to 52<!--> <!-->°C. The enzyme activity was inhibited by phenylmethane sulfonyl fluoride (PMSF) and ethylenediamine tetracetic acid (EDTA), indicating that it is a metal-dependent serine protease. It was found to be a direct acting plasmin like protein which efficiently cleaved the α-chain of fibrin(ogen), followed by β-chain and γ-chain. Three internal peptide sequences LASTANSGVLSGLLAGTVGGK; AYTSKSSVPSSVGLAR; LLDTGLNTAHSDFNR were determined by Q-TOF2. These results indicate no sequence similarities with other fibrinolytic enzymes suggesting it to be a novel enzyme. This fibrinolytic enzyme may be developed as a safe potential candidate for oral administration as a functional food additive or as a drug for prevention and/or treatment of thrombolytic diseases.</p></div>","PeriodicalId":16416,"journal":{"name":"Journal of Molecular Catalysis B-enzymatic","volume":"134 ","pages":"Pages 98-104"},"PeriodicalIF":0.0000,"publicationDate":"2016-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.molcatb.2016.10.006","citationCount":"16","resultStr":"{\"title\":\"Purification and characterization of a fibrinolytic enzyme from the food-grade fungus, Neurospora sitophila\",\"authors\":\"Xiao lan Liu,&nbsp;Narasimha kumar Kopparapu,&nbsp;Hong chen Zheng,&nbsp;Priti Katrolia,&nbsp;Yong ping Deng,&nbsp;Xi qun Zheng\",\"doi\":\"10.1016/j.molcatb.2016.10.006\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>A fibrinolytic protease was purified from the culture supernatant of a GRAS fungus, <em>Neurospora sitophila</em>. The enzyme displayed a molecular mass of 34<!--> <!-->kDa, as estimated by SDS-PAGE and gel filtration chromatography. The isoelectric point (pI) of the enzyme was 9.3<!--> <!-->±<!--> <!-->0.2 as determined by iso-electric focusing (IEF). It was maximally active at pH 7.6 and 41<!--> <!-->°C and displayed remarkable stability in a wide pH range (4–11) and up to 52<!--> <!-->°C. The enzyme activity was inhibited by phenylmethane sulfonyl fluoride (PMSF) and ethylenediamine tetracetic acid (EDTA), indicating that it is a metal-dependent serine protease. It was found to be a direct acting plasmin like protein which efficiently cleaved the α-chain of fibrin(ogen), followed by β-chain and γ-chain. Three internal peptide sequences LASTANSGVLSGLLAGTVGGK; AYTSKSSVPSSVGLAR; LLDTGLNTAHSDFNR were determined by Q-TOF2. These results indicate no sequence similarities with other fibrinolytic enzymes suggesting it to be a novel enzyme. This fibrinolytic enzyme may be developed as a safe potential candidate for oral administration as a functional food additive or as a drug for prevention and/or treatment of thrombolytic diseases.</p></div>\",\"PeriodicalId\":16416,\"journal\":{\"name\":\"Journal of Molecular Catalysis B-enzymatic\",\"volume\":\"134 \",\"pages\":\"Pages 98-104\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-12-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/j.molcatb.2016.10.006\",\"citationCount\":\"16\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Molecular Catalysis B-enzymatic\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1381117716301990\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"Chemical Engineering\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Molecular Catalysis B-enzymatic","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1381117716301990","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Chemical Engineering","Score":null,"Total":0}
引用次数: 16

摘要

从嗜sitophila神经孢子菌(Neurospora sitophila)的培养上清液中纯化出一种纤维蛋白溶解蛋白酶。经SDS-PAGE和凝胶过滤色谱分析,该酶的分子量为34 kDa。等电聚焦(IEF)测定酶的等电点(pI)为9.3±0.2。它在pH 7.6和41°C时具有最大的活性,在pH范围(4-11)和高达52°C时表现出显著的稳定性。该酶的活性被苯基甲烷磺酰氟(PMSF)和乙二胺四乙酸(EDTA)抑制,表明它是一种金属依赖性丝氨酸蛋白酶。发现它是一种直接作用的类纤溶酶蛋白,能有效地切割纤维蛋白(原)的α-链,其次是β链和γ链。三个内部肽序列LASTANSGVLSGLLAGTVGGK;AYTSKSSVPSSVGLAR;采用Q-TOF2法测定LLDTGLNTAHSDFNR。这些结果表明它与其他纤溶酶的序列没有相似性,提示它可能是一种新型酶。该纤溶酶可作为一种安全的潜在候选物,作为功能性食品添加剂口服给药,或作为预防和/或治疗溶栓性疾病的药物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Purification and characterization of a fibrinolytic enzyme from the food-grade fungus, Neurospora sitophila

Purification and characterization of a fibrinolytic enzyme from the food-grade fungus, Neurospora sitophila

A fibrinolytic protease was purified from the culture supernatant of a GRAS fungus, Neurospora sitophila. The enzyme displayed a molecular mass of 34 kDa, as estimated by SDS-PAGE and gel filtration chromatography. The isoelectric point (pI) of the enzyme was 9.3 ± 0.2 as determined by iso-electric focusing (IEF). It was maximally active at pH 7.6 and 41 °C and displayed remarkable stability in a wide pH range (4–11) and up to 52 °C. The enzyme activity was inhibited by phenylmethane sulfonyl fluoride (PMSF) and ethylenediamine tetracetic acid (EDTA), indicating that it is a metal-dependent serine protease. It was found to be a direct acting plasmin like protein which efficiently cleaved the α-chain of fibrin(ogen), followed by β-chain and γ-chain. Three internal peptide sequences LASTANSGVLSGLLAGTVGGK; AYTSKSSVPSSVGLAR; LLDTGLNTAHSDFNR were determined by Q-TOF2. These results indicate no sequence similarities with other fibrinolytic enzymes suggesting it to be a novel enzyme. This fibrinolytic enzyme may be developed as a safe potential candidate for oral administration as a functional food additive or as a drug for prevention and/or treatment of thrombolytic diseases.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Journal of Molecular Catalysis B-enzymatic
Journal of Molecular Catalysis B-enzymatic 生物-生化与分子生物学
CiteScore
2.58
自引率
0.00%
发文量
0
审稿时长
3.4 months
期刊介绍: Journal of Molecular Catalysis B: Enzymatic is an international forum for researchers and product developers in the applications of whole-cell and cell-free enzymes as catalysts in organic synthesis. Emphasis is on mechanistic and synthetic aspects of the biocatalytic transformation. Papers should report novel and significant advances in one or more of the following topics; Applied and fundamental studies of enzymes used for biocatalysis; Industrial applications of enzymatic processes, e.g. in fine chemical synthesis; Chemo-, regio- and enantioselective transformations; Screening for biocatalysts; Integration of biocatalytic and chemical steps in organic syntheses; Novel biocatalysts, e.g. enzymes from extremophiles and catalytic antibodies; Enzyme immobilization and stabilization, particularly in non-conventional media; Bioprocess engineering aspects, e.g. membrane bioreactors; Improvement of catalytic performance of enzymes, e.g. by protein engineering or chemical modification; Structural studies, including computer simulation, relating to substrate specificity and reaction selectivity; Biomimetic studies related to enzymatic transformations.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信