title: Cloning and partial sequencing of an operon encoding two Pseudomonas putida haloalkanoate dehalogenases of opposite stereospecificity.
abstract: We have cloned fragments of DNA (up to 13 kb), from Pseudomonas putida AJ1, that code for two stereospecific haloalkanoate dehalogenases. These enzymes are highly specific for D and L substrates. The two genes, designated hadD and hadL, have been isolated and independently expressed in Escherichia coli and P. putida hosts by using broad-host-range vectors. They are closely adjacent and inducible in what appears to be an operon with an upstream open reading frame of unknown function. Nucleotide sequence determination of hadD predicts a mature, cytoplasmic protein of 300 amino acid residues (molecular weight of 33,601). This has no significant homology with the L-specific haloalkanoate dehalogenases from Pseudomonas sp. strain CBS3 (B. Schneider, R. Muller, R. Frank, and F. Lingens, J. Bacteriol. 173:1530-1535, 1991) nor with any other known DNA or protein sequences.
authors.affiliation: Biotechnology Department, ICI Pharmaceuticals, Macclesfield, Cheshire, United Kingdom.
authors.forename: P T
authors.initials: PT
authors.lastname: Barth
authors.forename: L
authors.initials: L
authors.lastname: Bolton
authors.forename: J C
authors.initials: JC
authors.lastname: Thomson
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D005822:Genetic Vectors; D006867:Hydrolases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D009876:Operon; D010957:Plasmids; D016958:Pseudomonas putida / Q000235:genetics*; D015183:Restriction Mapping; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D004269:DNA, Bacterial; D009838:Oligodeoxyribonucleotides; D006867:Hydrolases; C034507:2-haloacid dehalogenase