Biomaterial Database

Purification and characterization of 4-chlorobenzoyl CoA dehalogenase from Arthrobacter sp. strain 4-CB1. 1994

G P Crooks, and S D Copley

Department of Chemistry and Biochemistry, University of Colorado at Boulder 80309.

4-Chlorobenzoyl coenzyme A dehalogenase was purified to homogeneity from Arthrobacter sp. strain 4-CB1 (previously known as Acinetobacter sp. strain 4-CB1), a bacterium isolated from PCB-contaminated soil. Purification was accomplished by four chromatographic steps, including a novel affinity chromatography step. 4-Chlorobenzoyl CoA dehalogenase is a homotetramer of 33-kDa subunits with an isoelectric point of 6.1. The enzyme is stable between pH 6.5 and 10. The optimum pH for kcat is pH 8. The enzyme is able to dehalogenate substrates bearing fluorine, chlorine, bromine and iodine in the 4-position, although the rate of dehalogenation of 4-fluorobenzoyl CoA is quite slow. The enzyme is specific for dehalogenation at the 4-position, as 3-chloro- and 2-chlorobenzoyl CoA are not dehalogenated. The N-terminal sequence of the Arthrobacter sp. strain 4-CB1 dehalogenase is almost identical to that of the 4-chlorobenzoyl CoA dehalogenase from Arthrobacter sp. strain SU and shows 30% identity to that from Pseudomonas sp. strain CBS-3.

UI	MeSH Term	Description	Entries
D007526	Isoelectric Point	The pH in solutions of proteins and related compounds at which the dipolar ions are at a maximum.	Isoelectric Points,Point, Isoelectric,Points, Isoelectric
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D008969	Molecular Sequence Data	Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.	Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D008970	Molecular Weight	The sum of the weight of all the atoms in a molecule.	Molecular Weights,Weight, Molecular,Weights, Molecular
D006863	Hydrogen-Ion Concentration	The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH	pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D006867	Hydrolases	Any member of the class of enzymes that catalyze the cleavage of the substrate and the addition of water to the resulting molecules, e.g., ESTERASES, glycosidases (GLYCOSIDE HYDROLASES), lipases, NUCLEOTIDASES, peptidases (PEPTIDE HYDROLASES), and phosphatases (PHOSPHORIC MONOESTER HYDROLASES). EC 3.	Hydrolase
D000595	Amino Acid Sequence	The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.	Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D001173	Arthrobacter	A genus of asporogenous bacteria isolated from soil that displays a distinctive rod-coccus growth cycle.
D013379	Substrate Specificity	A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.	Specificities, Substrate,Specificity, Substrate,Substrate Specificities
D017386	Sequence Homology, Amino Acid	The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.	Homologous Sequences, Amino Acid,Amino Acid Sequence Homology,Homologs, Amino Acid Sequence,Homologs, Protein Sequence,Homology, Protein Sequence,Protein Sequence Homologs,Protein Sequence Homology,Sequence Homology, Protein,Homolog, Protein Sequence,Homologies, Protein Sequence,Protein Sequence Homolog,Protein Sequence Homologies,Sequence Homolog, Protein,Sequence Homologies, Protein,Sequence Homologs, Protein