Jong Deog Kim , Satya Singh , Sachiko Machida , Young Yu , Chika Aoyagi , Yasushi Kawata , Kiyoshi Hayashi
{"title":"gilvus Cellvibrio c端5个氨基酸残基对β-葡萄糖苷酶折叠和稳定性的重要性","authors":"Jong Deog Kim , Satya Singh , Sachiko Machida , Young Yu , Chika Aoyagi , Yasushi Kawata , Kiyoshi Hayashi","doi":"10.1016/S0922-338X(98)80089-5","DOIUrl":null,"url":null,"abstract":"<div><p>To determine the role of the C-terminal region of <em>Cellvibrio gilvus</em> β-glucosidase, a deletion mutant was constructed lacking five amino acid residues (RGRAR), three of which were arginine, from the C-terminal end. The mutant, designated ΔRGRAR, could be folded into an active form when expressed with the molecular chaperons <span><math><mtext>GroEL</mtext><mtext>ES</mtext></math></span>. In comparison with the native enzyme, the optimum pH of the mutant ΔRGRAR shifted to the acidic region and the pH stability to the neutral region, while its heat stability decreased. No significant difference in the kinetic parameter <em>K</em><sub>m</sub> was observed. It was concluded that the RGRAR residues located at the C-terminal end are quite important for the stability of the enzyme and protein folding.</p></div>","PeriodicalId":15696,"journal":{"name":"Journal of Fermentation and Bioengineering","volume":"85 4","pages":"Pages 433-435"},"PeriodicalIF":0.0000,"publicationDate":"1998-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0922-338X(98)80089-5","citationCount":"11","resultStr":"{\"title\":\"Importance of five amino acid residues at C-terminal region for the folding and stability of β-glucosidase of Cellvibrio gilvus\",\"authors\":\"Jong Deog Kim , Satya Singh , Sachiko Machida , Young Yu , Chika Aoyagi , Yasushi Kawata , Kiyoshi Hayashi\",\"doi\":\"10.1016/S0922-338X(98)80089-5\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>To determine the role of the C-terminal region of <em>Cellvibrio gilvus</em> β-glucosidase, a deletion mutant was constructed lacking five amino acid residues (RGRAR), three of which were arginine, from the C-terminal end. The mutant, designated ΔRGRAR, could be folded into an active form when expressed with the molecular chaperons <span><math><mtext>GroEL</mtext><mtext>ES</mtext></math></span>. In comparison with the native enzyme, the optimum pH of the mutant ΔRGRAR shifted to the acidic region and the pH stability to the neutral region, while its heat stability decreased. No significant difference in the kinetic parameter <em>K</em><sub>m</sub> was observed. It was concluded that the RGRAR residues located at the C-terminal end are quite important for the stability of the enzyme and protein folding.</p></div>\",\"PeriodicalId\":15696,\"journal\":{\"name\":\"Journal of Fermentation and Bioengineering\",\"volume\":\"85 4\",\"pages\":\"Pages 433-435\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1998-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/S0922-338X(98)80089-5\",\"citationCount\":\"11\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Fermentation and Bioengineering\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0922338X98800895\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Fermentation and Bioengineering","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0922338X98800895","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Importance of five amino acid residues at C-terminal region for the folding and stability of β-glucosidase of Cellvibrio gilvus
To determine the role of the C-terminal region of Cellvibrio gilvus β-glucosidase, a deletion mutant was constructed lacking five amino acid residues (RGRAR), three of which were arginine, from the C-terminal end. The mutant, designated ΔRGRAR, could be folded into an active form when expressed with the molecular chaperons . In comparison with the native enzyme, the optimum pH of the mutant ΔRGRAR shifted to the acidic region and the pH stability to the neutral region, while its heat stability decreased. No significant difference in the kinetic parameter Km was observed. It was concluded that the RGRAR residues located at the C-terminal end are quite important for the stability of the enzyme and protein folding.