title: Enhanced and feedback-resistant gamma-glutamyl kinase activity of an Escherichia coli transformant carrying a mutated proB gene of Streptococcus thermophilus.
abstract: We used a PCR-based method to generate a single base pair mutation in the proB gene of Streptococcus thermophilus, which replaced an aspartic acid with a glycine residue at position 192 of the first proline biosynthetic enzyme gamma-glutamyl kinase. This was the first identified mutation in amino acid biosynthesis in S. thermophilus to our knowledge. The mutation caused an enhanced, feedback-resistant gamma-glutamyl kinase activity and conferred an analogue-resistant phenotype to an Escherichia coli transformant containing the mutated gene.
authors.affiliation: Department of General and Environmental Physiology, University Federico II, via Mezzocannone 16, 80134, Naples, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Massarelli
authors.forename: G
authors.initials: G
authors.lastname: Forlani
authors.forename: E
authors.initials: E
authors.lastname: Ricca
authors.forename: M
authors.initials: M
authors.lastname: De Felice
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001383:Azetidinecarboxylic Acid / Q000494:pharmacology; D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005246:Feedback; D008969:Molecular Sequence Data; D017851:Phosphotransferases (Carboxyl Group Acceptor) / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011392:Proline / Q000031:analogs & derivatives / Q000494:pharmacology; D013291:Streptococcus / Q000201:enzymology / Q000235:genetics*; D014169:Transformation, Bacterial
chemical_list: C002195:3,4-dehydroproline; D001383:Azetidinecarboxylic Acid; D011392:Proline; D017851:Phosphotransferases (Carboxyl Group Acceptor); C020650:glutamate 5-kinase