title: Engineering of microheterogeneity-resistant p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens.
abstract: By site-directed mutagenesis, Cys-116 was converted to Ser-116 in p-hydroxybenzoate hydroxylase (EC 1.14.13.2) from Pseudomonas fluorescens. In contrast to wild-type enzyme, the C116S mutant is no longer susceptible to oxidation by hydrogen peroxide and shows no reactivity towards 5,5'-dithiobis(2-nitrobenzoate). Crystals of the C116S mutant are isomorphous with the crystal form of wild-type enzyme. A difference electron density confirms the mutation made.
authors: Department of Biochemistry, Agricultural University, Wageningen, The Netherlands.
authors: K
authors: K
authors: Eschrich
authors: W J
authors: WJ
authors: van Berkel
authors: A H
authors: AH
authors: Westphal
authors: A
authors: A
authors: de Kok
authors: A
authors: A
authors: Mattevi
authors: G
authors: G
authors: Obmolova
authors: K H
authors: KH
authors: Kalk
authors: W G
authors: WG
authors: Hol
mesh_terms: D015109:4-Hydroxybenzoate-3-Monooxygenase / Q000037:antagonists & inhibitors / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D011487:Protein Conformation; D011549:Pseudomonas / Q000201:enzymology*; D013439:Sulfhydryl Reagents / Q000494:pharmacology
chemical_list: D009841:Oligonucleotides; D013439:Sulfhydryl Reagents; D015109:4-Hydroxybenzoate-3-Monooxygenase