{"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":null,"pages":null},"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\":null,\"pages\":null},\"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}
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.