[Protein engineering of uridine phosphorylase from Escherichia coli K-12. I. Cloning and expression of uridine phosphorylase genes from Klebsiella aerogenes and Salmonella typhimurium in E. coli].
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.
Veĭko
V P
VP
State Research Institute of Genetics and Selection of Industrial Microorganisms, Moscow, Russia.
Chebotaev
D V
DV
Ovcharova
I V
IV
Gul'ko
L B
LB
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D007711:Klebsiella pneumoniae / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D012486:Salmonella typhimurium / Q000201:enzymology / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D014543:Uridine Phosphorylase / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification
D009838:Oligodeoxyribonucleotides; D014543:Uridine Phosphorylase
Belkovaia inzheneriia uridinofosforilazy iz Escherichia coli K-12. I. Klonirovanie i ékspressiia v E. coli genov uridinofosforilaz iz Klebsiella aerogenes i Salmonella typhimurium.