Publications

• Enzymatic dehalogenation of certain brominated and chlorinated compounds.

  HEPPEL L.A., PORTERFIELD V.T.

  J Biol Chem 176:763-769(1948) [PubMed] [EuropePMC]

  This publication is cited by 2 other entries.

• Purification and properties of a new enzyme, DL-2-haloacid dehalogenase, from Pseudomonas sp.

  Motosugi K., Esaki N., Soda K.

  A new enzyme, DL-2-haloacid dehalogenase, was isolated and purified to homogeneity from the cells of Pseudomonas sp. strain 113. This enzyme catalyzed non-stereospecific dehalogenation of both of the optical isomers of 2-chloropropionate through an SN2 type of reaction; L- and D-lactates were form ... >> More

  A new enzyme, DL-2-haloacid dehalogenase, was isolated and purified to homogeneity from the cells of Pseudomonas sp. strain 113. This enzyme catalyzed non-stereospecific dehalogenation of both of the optical isomers of 2-chloropropionate through an SN2 type of reaction; L- and D-lactates were formed from D- and L-2-chloropropionates, respectively. The enzyme acted on 2-halogenated aliphatic carboxylic acids whose carbon chain lengths were less than five. It also dehalogenated trichloroacetate to form oxalate and showed maximum activity at pH 9.5. The Michaelis constants for substrates were as follows: 5.0 mM for monochloroacetate, 1.1 mM for L-2-chloropropionate, and 4.8 mM for D-2-chloropropionate. DL-2-Haloacid dehalogenase was inhibited by HgCl2, ZnSO4, and MnSO4, but was not affected by thiol reagents, such as p-chloromercuribenzoate and iodoacetamide. This enzyme had a molecular weight of about 68,000 and appeared to be composed of two subunits identical in molecular weight. << Less

  J Bacteriol 150:522-527(1982) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Purification and characterization of D-2-haloacid dehalogenase from Pseudomonas putida strain AJ1/23.

  Smith J.M., Harrison K., Colby J.

  A D-2-haloacid dehalogenase was isolated and purified to homogeneity from Pseudomonas putida strain AJ1/23. The enzyme catalysed the stereospecific dehalogenation of the D-isomer of 2-chloropropionate. Using a new ion-chromatograph assay, the enzyme was found to catalyse the dehalogenation of shor ... >> More

  A D-2-haloacid dehalogenase was isolated and purified to homogeneity from Pseudomonas putida strain AJ1/23. The enzyme catalysed the stereospecific dehalogenation of the D-isomer of 2-chloropropionate. Using a new ion-chromatograph assay, the enzyme was found to catalyse the dehalogenation of short-chain 2-halocarboxylic acids. Maximum enzyme activity occurred at pH 9.5 and 50 degrees C and the enzyme was insensitive to most -SH reagents. The enzyme has an Mr of about 135,000 and appears to be composed of four subunits of identical Mr. << Less

  J Gen Microbiol 136:881-886(1990) [PubMed] [EuropePMC]

• Stereospecificity of 2-monochloropropionate dehalogenation by the two dehalogenases of Pseudomonas putida PP3: evidence for two different dehalogenation mechanisms.

  Weightman A.J., Weightman A.L., Slater J.H.

  Pseudomonas putida PP3 grew on DL-2-monochloropropionate (2MCPA) with a release of chloride ions consistent with the dechlorination of both isomers. The organism grew on either D- or L-2MCPA. Dehalogenase activity in cell-free extracts showed that both D- and L-2MCPA were dehalogenated. Pseudomona ... >> More

  Pseudomonas putida PP3 grew on DL-2-monochloropropionate (2MCPA) with a release of chloride ions consistent with the dechlorination of both isomers. The organism grew on either D- or L-2MCPA. Dehalogenase activity in cell-free extracts showed that both D- and L-2MCPA were dehalogenated. Pseudomonas putida PP3 contains two dehalogenases, and studies with the separated enzymes revealed that the fraction I enzyme used both D- and L-2MCPA, the rate of dechlorination of L-2MCPA being 80% of the rate of D-2MCPA dechlorination. The product of the reaction, lactate, retained the same optical configuration as the substrate provided. The fraction II dehalogenase also dechlorinated D- and L-2MCPA, with the same difference in rates as for the fraction I dehalogenase, but the lactates produced were of the opposite configuration to their precursors. The two dehalogenases showed further differences with respect to inhibition by two sulphydryl-blocking agents, N-ethylmaleimide and p-chloromercuribenzoate. Fraction I dehalogenase was considerably more sensitive to these two reagents compared with the fraction II dehalogenase. Dithiothreitol partially protected the fraction I dehalogenase from N-ethylmaleimide inhibition. The results are discussed in terms of the possible evolutionary relationships of the two dehalogenases. << Less

  J Gen Microbiol 128:1755-1762(1982) [PubMed] [EuropePMC]

  This publication is cited by 3 other entries.

• Cloning, sequencing and expression in Escherichia coli of two Rhizobium sp. genes encoding haloalkanoate dehalogenases of opposite stereospecificity.

  Cairns S.S., Cornish A., Cooper R.A.

  A 6.5-kp EcoRI fragment of genomic DNA from a Rhizobium sp. cloned into pUC19 was able to endow Escherichia coli K-12 with the novel ability to grow at the expense of 2-chloropropionic acid. Subcloning showed that this property was a consequence of two dehalogenases encoded on a 2.2-kb PstI fragme ... >> More

  A 6.5-kp EcoRI fragment of genomic DNA from a Rhizobium sp. cloned into pUC19 was able to endow Escherichia coli K-12 with the novel ability to grow at the expense of 2-chloropropionic acid. Subcloning showed that this property was a consequence of two dehalogenases encoded on a 2.2-kb PstI fragment. Further subcloning of the PstI fragment led to two constructs that encoded, separately, dehalogenase activity that acted stereospecifically on D-2-chloropropionic acid and L-2-cloropropionic acid, respectively. The genes encoding these two stereospecific dehalogenases have been sequenced and shown to be separated by 177 bp of non-coding DNA. Expression of the dehalogenase genes involved the vector promoter, suggesting that the anticipated Rhizobium sp. regulatory sequences were not functional in E. coli. Comparison of the deduced amino acid sequences of the two dehalogenases (18% identity) indicated with any other 2-chloropropionic acid dehalogenase studied so far. << Less

  Eur. J. Biochem. 235:744-749(1996) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Bacterial assimilation of D- and L-2-chloropropionates and occurrence of a new dehalogenase.

  Motosugi K., Esaki N., Soda K.

  The occurrence of a new bacterial dehalogenase acting on both the optical isomers of 2-halogenated alkanoic acids was demonstrated. When the haloalkanoic acid-utilizing bacteria were screened in a medium containing DL-2-chloropropionate as a sole carbon source, two types of bacteria were isolated: ... >> More

  The occurrence of a new bacterial dehalogenase acting on both the optical isomers of 2-halogenated alkanoic acids was demonstrated. When the haloalkanoic acid-utilizing bacteria were screened in a medium containing DL-2-chloropropionate as a sole carbon source, two types of bacteria were isolated: (1) a few strains utilizing both D- and L-isomers of 2-chloropropionate and (2) strains utilizing only the L-isomer. A dehalogenating enzyme was obtained from the cells of Pseudomonas sp. which is able to utilize both isomers. The crude enzyme catalyzed the dehalogenation of D- and L-2-chloropropionates to yield L- and D-isomers of lactate, respectively. The enzyme showed the same pH optimum and heat inactivation rate for the D- and L-isomers. Apparent Km values for D- and L-2-chloropropionates were 4.5 and 1.0mM, respectively. The enzyme acted specifically on 2-haloalkanoic acids. Activity staining of disc-gels electrophoresed with the crude enzyme preparation showed that the dehalogenation of D- and L-2-chloropropionates, monochloroacetate, dichloroacetate, 2,2-dichloropropionate, and DL-2-chlorobutyrate is due to a single protein. << Less

  Arch Microbiol 131:179-183(1982) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Purification and characterization of thermostable and nonthermostable 2-haloacid dehalogenases with different stereospecificities from Pseudomonas sp. strain YL.

  Liu J.Q., Kurihara T., Hasan A.K., Nardi-Dei V., Koshikawa H., Esaki N., Soda K.

  Two novel hydrolytic dehalogenases, thermostable L-2-haloacid dehalogenase (L-DEX) inducibly synthesized by 2-chloropropionate (2-CPA) and nonthermostable DL-2-haloacid dehalogenase (DL-DEX) induced by 2-chloroacrylate, were purified to homogeneity from Pseudomonas sp. strain YL. DL-DEX consisted ... >> More

  Two novel hydrolytic dehalogenases, thermostable L-2-haloacid dehalogenase (L-DEX) inducibly synthesized by 2-chloropropionate (2-CPA) and nonthermostable DL-2-haloacid dehalogenase (DL-DEX) induced by 2-chloroacrylate, were purified to homogeneity from Pseudomonas sp. strain YL. DL-DEX consisted of a monomer with a molecular weight of about 36,000 and catalyzed the dehalogenation of L and D isomers of 2-CPA to produce D- and L-lactates, respectively. It acted on 2-haloalkanoic acids with a carbon chain length of 2 to 4. The maximum activity on DL-2-CPA was found at pH 10.5 and 45 degrees C. L-DEX, composed of two subunits with identical molecular weights of 27,000, catalyzes the dehalogenation of L-2-haloalkanoic acids to produce the corresponding D-2-hydroxyalkanoic acids. The enzyme acts not only on short-carbon-chain 2-haloacids such as monochloroacetate and monoiodoacetate in aqueous solution but also on long-carbon-chain 2-haloacids such as 2-bromohexadecanoate in n-heptane. L-DEX is thermostable: it retained its full activity upon heating at 60 degrees C for 30 min. The pH and temperature optima for dehalogenation of L-2-CPA were 9.5 and 65 degrees C, respectively. L-DEX was strongly inhibited by modification of carboxyl groups with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and Woodward reagent K, but DL-DEX was not. << Less

  Appl Environ Microbiol 60:2389-2393(1994) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.