Lack of homology between two haloacetate dehalogenase genes encoded on a plasmid from Moraxella sp. strain B

Abstract

Two genes encoding haloacetate dehalogenases, H-1 and H-2, are closely linked on a plasmid from Moraxellu sp. strain B. H-1 predominantly acts on fluoroacetate, but H-2 does not. To elucidate the molecular relationship between the two enzymes, we compared their structural genes. Two restriction fragments of the plasmid DNA were subcloned on M13 phages and their nucleotide sequences were determined. The sequence of each fragment contained an open reading frame that was identified as the structural gene for each of the two dehalogenases on the basis of the following criteria; N-terminal amino acid sequence, amino acid composition, and molecular mass. The genes for H-1 and H-2, designated &hHl and khH2, respectively, had Merent sizes (885 bp and 675 bp) and G + C contents (58.3% and 53.4%). Sequence analysis revealed no homology between the two genes. We concluded that the dehalogenases H-1 and H-2 have no enzyme-evolutionary relationship. The deduced amino acid sequence of the &hHl gene showed significant similarity to those of three hydrolases of Pseudomonasputida and a haloalkane dehalogenase of Xanthobacter rurtotrophicus. The &hH2 coding region was sandwiched between two repeated sequences about 1.8 kb long, which might play a part in the frequent spontaneous deletion of &hH2 from the plasmid.

Differential Expression of a Novel Gene in Response to Coronatine, Methyl Jasmonate, and Wounding in the Coi1Mutant of Arabidopsis1

“…The microbial dienelactone hydrolase hydrolyzes dienelactone to maleylacetate and has esterase activity toward other substrates (Pathak et al, 1991;Beveridge and Ollis, 1995). Haloacetate dehalogenase acts specifically on halogenated acetates to yield glycolate as a carbon source in microorganisms (Kawasaki et al, 1992). Epoxide hydrolase catalyzes the conversion of epoxides to diols.…”

Cloning and Sequence Analysis of a Plasmid-encoded 2-Haloacid Dehalogenase Gene fromPseudomonas putidaNo. 109

“…It has been reported that there is no homology between dehH1 and dehH2. 4 ) H-2 has less activity against 2-chloropropionate than H-I09 has, but other catalytic properties are similar between H-2 and H-I09. The length of dehH2 is 675bp that is identical with that of dehH109, and dehH2 and dehH109 had 58% similarity in the nucleotide sequence and 51 % similarity in the amino acid sequence.…”

Repositioning the Catalytic Triad Aspartic Acid of Haloalkane Dehalogenase: Effects on Stability, Kinetics, and Structure

“…Moraxella B (DehH1, Genbank/EMBL Accession Number D90422; Kawasaki et al, 1992), tetrachlorocyclohexadiene dehalogenase from Sphingomonas paucimobilis (LinB, D14594; Nagata et al, 1993), and 1-chloroalkane halidohydrolase from Rhodococcus rhodochrous NCIMB13064 (DhaA, L49435; Kulakova et al, 1997) showed that they have considerable homology with DhlA (M26950), suggesting FIGURE 2: Sequence alignment of haloalkane dehalogenase from Xanthobacter autotrophicus GJ10 (DhlA; Janssen et al, 1989), haloacid dehalogenase from Moraxella B (DehH1; Kawasaki et al, 1992), tetrachlorocyclohexadiene dehalogenase from Sphingomonas paucimobilis (LinB; Nagata et al, 1993), and 1-chloroalkane halidohydrolase from Rhodococcus rhodochrous NCIMB13064 (DhaA; Kulakova et al, 1997). The sequence comparison was carried out using the program DNASTAR (DNASTAR Inc., Madison, WI).…”