Protein engineering of uridine phosphorylase from Escherichia coli K-12. I. Cloning and expression in E-coli of genes for uridine phosphorylases from Klebsiella aerogenes and Salmonella typhimurium

作者：Veiko VP^*; Chebotarev DV; Ovcharova IV; Gul'ko LB

来源：Bioorganicheskaya Khimiya, 1998, 24(5): 381-387.

• uridine phosphorylase
• udp genes
• site-directed mutagenesis
• cloning
• PCR
• Escherichia coli
• Klebsiella aerogenes
• Salmonella typhimurium

Genes of uridine phosphorylases (udp) from Klebsiella aerogenes and Salmonella typhimurium were cloned and expressed. Highly effective producer strains of the corresponding proteins were constructed. Enzymic properties of the UPases obtained were studied and compared with those from the Escherichia coli enzyme. Mutant forms of UPase from E; coli (D5E, D5N, D5A) were prepared by site-directed mutagenesis techniques. It was shown that the Asp5 residue plays an insignificant role in the formation of the active form of the protein.

• 出版日期1998-5