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

Weightman, Andrew John ORCID: https://orcid.org/0000-0002-6671-2209, Weightman, Alison Lesley ORCID: https://orcid.org/0000-0001-5210-3798 and Slater, J.Howard 1982. Stereospecificity of 2-monochloropropionate dehalogenation by the two dehalogenases of pseudomonas putida PP3: evidence for two different dehalogenation mechanisms. Journal of General Microbiology 128 (8) , pp. 1755-1762. 10.1099/00221287-128-8-1755

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Abstract

SUMMARY: 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.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences Academic & Student Support Service
Publisher:	Society for General Microbiology
ISSN:	0022-1287
Last Modified:	17 Oct 2022 10:06
URI:	https://orca.cardiff.ac.uk/id/eprint/6800

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