title: Purification of histidase from Streptomyces griseus and nucleotide sequence of the hutH structural gene.
abstract: Histidine ammonia-lyase (histidase) was purified to homogeneity from vegetative mycelia of Streptomyces griseus. The enzyme was specific for L-histidine and showed no activity against the substrate analog, D-histidine. Histidinol phosphate was a potent competitive inhibitor. Histidase displayed saturation kinetics with no detectable sigmoidal response. Neither thiol reagents nor a variety of divalent cations had any effect on the activity of the purified enzyme. High concentrations of potassium cyanide inactivated histidase in the absence of its substrate or histidinol phosphate, suggesting that, as in other histidases, dehydroalanine plays an important role in catalysis. The N-terminal amino acid sequence of histidase was used to construct a mixed oligonucleotide probe to identify and clone the histidase structural gene, hutH, from genomic DNA of the wild-type strain of S. griseus. The cloned DNA restored the ability of a histidase structural gene mutant to grow on L-histidine as the sole nitrogen source. The deduced amino acid sequence of hutH shows significant relatedness with histidase from bacteria and a mammal as well as phenylalanine ammonia-lyase from plants and fungi.
authors.affiliation: Department of Microbiology, Ohio State University, Columbus 43210-1292.
authors.forename: P C
authors.initials: PC
authors.lastname: Wu
authors.forename: T A
authors.initials: TA
authors.lastname: Kroening
authors.forename: P J
authors.initials: PJ
authors.lastname: White
authors.forename: K E
authors.initials: KE
authors.lastname: Kendrick
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003486:Cyanides / Q000494:pharmacology; D006638:Histidine Ammonia-Lyase / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D006641:Histidinol / Q000031:analogs & derivatives; D008969:Molecular Sequence Data; D012045:Regulatory Sequences, Nucleic Acid; D012689:Sequence Homology, Nucleic Acid; D013305:Streptomyces griseus / Q000201:enzymology / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D003486:Cyanides; D006641:Histidinol; D006638:Histidine Ammonia-Lyase