Hao Li, Ling-Yan Wang, Gui-Yun Xu, Yang Chen, Rong Jiang, Yuan Li
{"title":"Cloning, expression and characterization of the Ste6 gene encoding a UDP-glucose dehydrogenase in Streptomyces.","authors":"Hao Li, Ling-Yan Wang, Gui-Yun Xu, Yang Chen, Rong Jiang, Yuan Li","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>Streptomyces sp. 139 was identified to produce a new exopolysaccharide Ebosin (139A) with antirheumatic arthritis activity in vivo. The Ebosin biosynthesis gene cluster(31.3 kb; GenBank Accession Number: AY131229) containing 22 ORFs (ste1-ste22) of Streptomyces sp. 139 had been reported previously. In this paper,we present experimental evidence for the identity of the ste6 gene product as a UDP-glucose dehydrogenase (UDPGDH). With pET-30a as vector,the gene was cloned and expressed in Escherichia coli BL21 (DE3). The expressed protein was purified to homogeneity by His-Bind resin affinity chromatograpy and it was able to catalyze UDP-glucose to UDP-glucuronic acid. To evaluate the function of ste6, the gene was disrupted by a single-crossover homologous recombination event and the result showed that ste6 is required in Ebosin biosynthesis.</p>","PeriodicalId":23770,"journal":{"name":"Yi chuan xue bao = Acta genetica Sinica","volume":"32 11","pages":"1213-20"},"PeriodicalIF":0.0000,"publicationDate":"2005-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Yi chuan xue bao = Acta genetica Sinica","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Streptomyces sp. 139 was identified to produce a new exopolysaccharide Ebosin (139A) with antirheumatic arthritis activity in vivo. The Ebosin biosynthesis gene cluster(31.3 kb; GenBank Accession Number: AY131229) containing 22 ORFs (ste1-ste22) of Streptomyces sp. 139 had been reported previously. In this paper,we present experimental evidence for the identity of the ste6 gene product as a UDP-glucose dehydrogenase (UDPGDH). With pET-30a as vector,the gene was cloned and expressed in Escherichia coli BL21 (DE3). The expressed protein was purified to homogeneity by His-Bind resin affinity chromatograpy and it was able to catalyze UDP-glucose to UDP-glucuronic acid. To evaluate the function of ste6, the gene was disrupted by a single-crossover homologous recombination event and the result showed that ste6 is required in Ebosin biosynthesis.