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

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Standard

Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene. / van der Ploeg, J; Van Hall, Gerrit; Janssen, D B.

I: Journal of Bacteriology, Bind 173, Nr. 24, 1991, s. 7925-33.

Publikation: Bidrag til tidsskriftTidsskriftartikelfagfællebedømt

Harvard

van der Ploeg, J, Van Hall, G & Janssen, DB 1991, 'Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene', Journal of Bacteriology, bind 173, nr. 24, s. 7925-33.

APA

van der Ploeg, J., Van Hall, G., & Janssen, D. B. (1991). Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene. Journal of Bacteriology, 173(24), 7925-33.

Vancouver

van der Ploeg J, Van Hall G, Janssen DB. Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene. Journal of Bacteriology. 1991;173(24):7925-33.

Author

van der Ploeg, J ; Van Hall, Gerrit ; Janssen, D B. / Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene. I: Journal of Bacteriology. 1991 ; Bind 173, Nr. 24. s. 7925-33.

Bibtex

@article{d74138b04f7411de87b8000ea68e967b,
title = "Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene",
abstract = "The haloacid dehalogenase of the 1,2-dichloroethane-utilizing bacterium Xanthobacter autotrophicus GJ10 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)2SO4 fractionation, DEAE-cellulose chromatography, and hydroxylapatite chromatography. The molecular mass of the protein was 28 kDa as determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 36 kDa as determined with gel filtration on Superose 12 fast protein liquid chromatography. The enzyme was active with 2-halogenated carboxylic acids and converted only the L-isomer of 2-chloropropionic acid with inversion of configuration to produce D-lactate. The activity of the enzyme was not readily influenced by thiol reagents. The gene encoding the haloacid dehalogenase (dhlB) was cloned and could be allocated to a 6.5-kb EcoRI-BglII fragment. Part of this fragment was sequenced, and the dhlB open reading frame was identified by comparison with the N-terminal amino acid sequence of the protein. The gene was found to encode a protein of 27,433 Da that showed considerable homology (60.5 and 61.0% similarity) with the two other haloacid dehalogenases sequenced to date but not with the haloalkane dehalogenase from X. autotrophicus GJ10.",
author = "{van der Ploeg}, J and {Van Hall}, Gerrit and Janssen, {D B}",
note = "Keywords: Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA, Bacterial; Electrophoresis, Polyacrylamide Gel; Genes, Bacterial; Gram-Negative Aerobic Bacteria; Hydrogen-Ion Concentration; Hydrolases; Molecular Sequence Data; Mutation; Restriction Mapping; Sequence Alignment; Substrate Specificity",
year = "1991",
language = "English",
volume = "173",
pages = "7925--33",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "24",

}

RIS

TY - JOUR

T1 - Characterization of the haloacid dehalogenase from Xanthobacter autotrophicus GJ10 and sequencing of the dhlB gene

AU - van der Ploeg, J

AU - Van Hall, Gerrit

AU - Janssen, D B

N1 - Keywords: Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA, Bacterial; Electrophoresis, Polyacrylamide Gel; Genes, Bacterial; Gram-Negative Aerobic Bacteria; Hydrogen-Ion Concentration; Hydrolases; Molecular Sequence Data; Mutation; Restriction Mapping; Sequence Alignment; Substrate Specificity

PY - 1991

Y1 - 1991

N2 - The haloacid dehalogenase of the 1,2-dichloroethane-utilizing bacterium Xanthobacter autotrophicus GJ10 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)2SO4 fractionation, DEAE-cellulose chromatography, and hydroxylapatite chromatography. The molecular mass of the protein was 28 kDa as determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 36 kDa as determined with gel filtration on Superose 12 fast protein liquid chromatography. The enzyme was active with 2-halogenated carboxylic acids and converted only the L-isomer of 2-chloropropionic acid with inversion of configuration to produce D-lactate. The activity of the enzyme was not readily influenced by thiol reagents. The gene encoding the haloacid dehalogenase (dhlB) was cloned and could be allocated to a 6.5-kb EcoRI-BglII fragment. Part of this fragment was sequenced, and the dhlB open reading frame was identified by comparison with the N-terminal amino acid sequence of the protein. The gene was found to encode a protein of 27,433 Da that showed considerable homology (60.5 and 61.0% similarity) with the two other haloacid dehalogenases sequenced to date but not with the haloalkane dehalogenase from X. autotrophicus GJ10.

AB - The haloacid dehalogenase of the 1,2-dichloroethane-utilizing bacterium Xanthobacter autotrophicus GJ10 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)2SO4 fractionation, DEAE-cellulose chromatography, and hydroxylapatite chromatography. The molecular mass of the protein was 28 kDa as determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 36 kDa as determined with gel filtration on Superose 12 fast protein liquid chromatography. The enzyme was active with 2-halogenated carboxylic acids and converted only the L-isomer of 2-chloropropionic acid with inversion of configuration to produce D-lactate. The activity of the enzyme was not readily influenced by thiol reagents. The gene encoding the haloacid dehalogenase (dhlB) was cloned and could be allocated to a 6.5-kb EcoRI-BglII fragment. Part of this fragment was sequenced, and the dhlB open reading frame was identified by comparison with the N-terminal amino acid sequence of the protein. The gene was found to encode a protein of 27,433 Da that showed considerable homology (60.5 and 61.0% similarity) with the two other haloacid dehalogenases sequenced to date but not with the haloalkane dehalogenase from X. autotrophicus GJ10.

M3 - Journal article

C2 - 1744048

VL - 173

SP - 7925

EP - 7933

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 24

ER -

ID: 12485061