Purification and properties of haloacetate halidohydrolase specified by plasmid from Moraxella sp. strain B.

Kawasaki, Haruhiko; Tone, Noriko; Tonomura, Kenzo

doi:10.1271/bbb1961.45.35

Cited by 44 publications

(28 citation statements)

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“…Dehalogenases produced by E. coli clones harbouring pBREFl and pBRSG2 were purified to homogeneity by the procedures described for the purification of H-1 and H-2 from Moraxella sp. B (Kawasaki et al, 1981 a). Cells grown in LB broth containing 30pg ampicillin ml-* were ruptured with an ultrasonic disintegrator.…”

Section: Methodsmentioning

confidence: 99%

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

Kawasaki

Tsuda

Matsushita

et al. 1992

Journal of General Microbiology

107

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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.

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Section: Methodsmentioning

confidence: 99%

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

Kawasaki

Tsuda

Matsushita

et al. 1992

Journal of General Microbiology

107

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“…3.8.1.2), a number of enzymes have been purified and characterized (14,16,17,19,22,29,34). They have been divided in different classes according to their substrate specificity (9), electrophoretic mobility on polyacrylamide gels (9, 36), and stereospecific action on 2-monochloropropionic acid (2-MCPA) (36).…”

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confidence: 99%

Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene

1991

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The haloacid dehalogenase of the 1,2-dichloroethane-utilizing bacterium Xanthobacter autotrophicus GJ1O was purified from a mutant with an eightfold increase in expression of the enzyme. The mutant was obtained by selecting for enhanced resistance to monobromoacetate. The enzyme was purified through (NH4)2S04 fractionation, DEAE-cellulose chromatography, and hydroxylapatite chromatography. The Hydrolytic dehalogenases are key enzymes in the detoxification of aliphatic halogenated hydrocarbons. They catalyze the cleavage of carbon-halogen bonds through a nucleophilic substitution by water to yield an alcohol. At least two distinct groups can be recognized with respect to their substrate ranges: haloalkane dehalogenases hydrolyze halogenated alkanes, whereas haloacid dehalogenases are active with short-chain 2-halogenated carboxylic acids.Of the 2-haloacid dehalogenases (E.C. 3.8.1.2), a number of enzymes have been purified and characterized (14,16,17,19,22,29,34). They have been divided in different classes according to their substrate specificity (9), electrophoretic mobility on polyacrylamide gels (9, 36), and stereospecific action on 2-monochloropropionic acid (2-MCPA) (36). Four different types of dehalogenation of 2-MCPA can be recognized. Two of these are represented by enzymes that are active with only L-or D-2-MCPA, giving products with inverted configuration at the chiral carbon atom. The other two act on both isomers, one with inversion of configuration and the other with retention of configuration.Previous studies have shown the presence of more than one haloacid dehalogenase in the same organism (9, 16), with only minor differences in substrate specificities. It has been suggested that these isoenzymes have arisen by gene duplication and subsequent divergent evolution (9).We are interested in the relation between structure, enzymatic mechanism, and evolution of dehalogenases. To study these aspects, we started to investigate the haloacid dehalogenase of Xanthobacter autotrophicus GJ1O. This bacterium was isolated on 1,2-dichloroethane as the sole carbon and energy source (13 In a previous paper, we reported the cloning of both dehalogenase genes from GJ10 (12). The haloalkane dehalogenase has been purified (15) and crystallized (25), and its sequence (12) and tertiary structure (5) have been determined. The haloacid dehalogenase gene has been cloned on a 10-kb fragment in the broad-host-range cosmid vector pLAFR1 (12). Here, we describe the purification and characterization of the haloacid dehalogenase from an overproducing mutant of GJ10 and the sequence of the gene encoding this enzyme. The properties of the enzyme are compared with those of other haloacid dehalogenases and haloalkane dehalogenase. MATERIALS AND METHODSGrowth conditions. Strains and plasmids used are listed in

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“…Some halogenated compounds are catabolized through enzymatic dehalogenation reactions by several bacteria (10-12, 25, 28, 31, 32, 36). Many investigations have been focused on the enzymatic cleavage of the carbonhalogen bonds (4,6,9,14,15,17,18,26,27,29,33).…”

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confidence: 99%

Resolution and some properties of enzymes involved in enantioselective transformation of 1,3-dichloro-2-propanol to (R)-3-chloro-1,2-propanediol by Corynebacterium sp. strain N-1074

Nakamura

Nagasawa

et al. 1992

J Bacteriol

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Purification and properties of haloacetate halidohydrolase specified by plasmid from Moraxella sp. strain B.

Cited by 44 publications

References 1 publication

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

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

Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene

Resolution and some properties of enzymes involved in enantioselective transformation of 1,3-dichloro-2-propanol to (R)-3-chloro-1,2-propanediol by Corynebacterium sp. strain N-1074

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