Cloning and sequence analysis of a plasmid-encoded 2-haloacid dehalogenase gene from Pseudomonas putida No. 109.
The 2-haloacid dehalogenase of Pseudomonas putida No. 109 was mediated by a 74-kb conjugative plasmid, which was transferred by mating into Pseudomonas and Escherichia coli and there expressed the dehalogenase. A 2.8-kb EcoRI-fragment generated from the plasmid was cloned and sequenced. The dehalogenase gene (dehH109) was identified by comparison with the N-terminal amino acid sequence and the molecular weight of the enzyme protein. The gene dehH109 coded for a 224-amino acid protein of M(r) 25,231, which showed significant homology to the other four L-specific 2-haloacid dehalogenases from Pseudomonas sp. CBS3, P. putida AJ1, and Xanthobacter autotrophicus GJ10 and also to the haloacetate dehalogenase H-2 from Moraxella sp. strain B, but no homology with another haloacetate dehalogenase H-1 and the D-specific 2-haloacid dehalogenase from P. putida AJ1.
Kawasaki
H
H
Department of Agricultural Chemistry, University of Osaka Prefecture, Japan.
Toyama
T
T
Maeda
T
T
Nishino
H
H
Tonomura
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics; D017353:Gene Deletion; D006867:Hydrolases / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D016958:Pseudomonas putida / Q000201:enzymology* / Q000235:genetics; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D006867:Hydrolases; C034507:2-haloacid dehalogenase