裂叶菌Bcc26414纤维素二糖脱氢酶的纯化及特性研究

Vinita Yadav, Sadhana Nighojkar
{"title":"裂叶菌Bcc26414纤维素二糖脱氢酶的纯化及特性研究","authors":"Vinita Yadav, Sadhana Nighojkar","doi":"10.13005/bbra/3141","DOIUrl":null,"url":null,"abstract":"CDH, an enzyme produced by wood-decomposing fungi, has diverse applications in biosensor production, bioremediation, and biomedical industries. In this study, CDH from Schizophyllum commune BCC26414 was purified using ammonium sulfate precipitation, DEAE-cellulose chromatography, and Sephadex G-200 chromatography. The purification fold achieved was 65.81 with a specific activity of 1612.34 U/mg. The purity and molecular weight of CDH was confirmed using native and SDS PAGE. Optimal temperature and pH were found to be 30°C and 5, respectively. The purified CDH exhibited stability over a wide pH range (3.5 to 6.5) for 24 hrs and retained complete activity at 40°C, with reduced activity at 50°C when observed for 150 min. KCl, MgSO4, ZnSO4, and NiCl2 at a concentration of 5 mM enhanced CDH activity and HgCl2 and CuSO4 inhibited the enzyme activity. The kinetic constants, Km and Vmax of CDH for lactose were observed to be 125 mM and 13.26 U/ml, respectively. The purified CDH may be utilized commercially in various applications.","PeriodicalId":9032,"journal":{"name":"Biosciences, Biotechnology Research Asia","volume":"469 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2023-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Cellobiose Dehydrogenase from Schizophyllum commune Bcc26414: Purification and Characterization\",\"authors\":\"Vinita Yadav, Sadhana Nighojkar\",\"doi\":\"10.13005/bbra/3141\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"CDH, an enzyme produced by wood-decomposing fungi, has diverse applications in biosensor production, bioremediation, and biomedical industries. In this study, CDH from Schizophyllum commune BCC26414 was purified using ammonium sulfate precipitation, DEAE-cellulose chromatography, and Sephadex G-200 chromatography. The purification fold achieved was 65.81 with a specific activity of 1612.34 U/mg. The purity and molecular weight of CDH was confirmed using native and SDS PAGE. Optimal temperature and pH were found to be 30°C and 5, respectively. The purified CDH exhibited stability over a wide pH range (3.5 to 6.5) for 24 hrs and retained complete activity at 40°C, with reduced activity at 50°C when observed for 150 min. KCl, MgSO4, ZnSO4, and NiCl2 at a concentration of 5 mM enhanced CDH activity and HgCl2 and CuSO4 inhibited the enzyme activity. The kinetic constants, Km and Vmax of CDH for lactose were observed to be 125 mM and 13.26 U/ml, respectively. The purified CDH may be utilized commercially in various applications.\",\"PeriodicalId\":9032,\"journal\":{\"name\":\"Biosciences, Biotechnology Research Asia\",\"volume\":\"469 1\",\"pages\":\"0\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-10-05\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biosciences, Biotechnology Research Asia\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.13005/bbra/3141\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biosciences, Biotechnology Research Asia","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.13005/bbra/3141","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

摘要

CDH是一种由木材分解真菌产生的酶,在生物传感器生产、生物修复和生物医学工业中有着广泛的应用。本研究采用硫酸铵沉淀法、deae -纤维素层析法和Sephadex G-200层析法对Schizophyllum commune BCC26414中的CDH进行纯化。纯化倍数为65.81,比活性为1612.34 U/mg。CDH的纯度和分子量通过native和SDS PAGE进行了验证。最佳温度为30℃,pH为5℃。纯化后的CDH在较宽的pH范围(3.5 ~ 6.5)内表现出24小时的稳定性,在40°C下保持完全的活性,在50°C下观察150分钟后活性降低。KCl、MgSO4、ZnSO4和NiCl2在5 mM浓度下增强CDH活性,HgCl2和CuSO4抑制酶活性。CDH对乳糖的动力学常数、Km和Vmax分别为125 mM和13.26 U/ml。纯化后的CDH可用于各种商业用途。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Cellobiose Dehydrogenase from Schizophyllum commune Bcc26414: Purification and Characterization
CDH, an enzyme produced by wood-decomposing fungi, has diverse applications in biosensor production, bioremediation, and biomedical industries. In this study, CDH from Schizophyllum commune BCC26414 was purified using ammonium sulfate precipitation, DEAE-cellulose chromatography, and Sephadex G-200 chromatography. The purification fold achieved was 65.81 with a specific activity of 1612.34 U/mg. The purity and molecular weight of CDH was confirmed using native and SDS PAGE. Optimal temperature and pH were found to be 30°C and 5, respectively. The purified CDH exhibited stability over a wide pH range (3.5 to 6.5) for 24 hrs and retained complete activity at 40°C, with reduced activity at 50°C when observed for 150 min. KCl, MgSO4, ZnSO4, and NiCl2 at a concentration of 5 mM enhanced CDH activity and HgCl2 and CuSO4 inhibited the enzyme activity. The kinetic constants, Km and Vmax of CDH for lactose were observed to be 125 mM and 13.26 U/ml, respectively. The purified CDH may be utilized commercially in various applications.
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
自引率
0.00%
发文量
0
×
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