1887

Abstract

PP3 grew on -2-monochloropropionate (2MCPA) with a release of chloride ions consistent with the dechlorination of both isomers. The organism grew on either -or -2MCPA. Dehalogenase activity in cell-free extracts showed that both -and -2MCPA were dehalogenated. PP3 contains two dehalogenases, and studies with the separated enzymes revealed that the fraction I enzyme used both -and -2MCPA, the rate of dechlorination of -2MCPA being 80% of the rate of -2MCPA dechlorination. The product of the reaction, lactate, retained the same optical configuration as the substrate provided. The fraction II dehalogenase also dechlorinated -and -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, -ethylmaleimide and -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 -ethylmaleimide inhibition. The results are discussed in terms of the possible evolutionary relationships of the two dehalogenases.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-128-8-1755
1982-08-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/128/8/mic-128-8-1755.html?itemId=/content/journal/micro/10.1099/00221287-128-8-1755&mimeType=html&fmt=ahah

References

  1. Berry E. I. M., Allison N., Skinner A. J., Cooper R. A. 1979; Degradation of the selective herbicide 2,2-dichloropropionate (Dalapon) by a soil bacterium. Journal of General Microbiology 110:39–45
    [Google Scholar]
  2. Davies J. L., Evans W. C. 1962; The elimination of halide ions from aliphatic halogen-substituted organic acids by an enzyme preparation from Pseudomonas dehalogens. Biochemical Journal 82:50P
    [Google Scholar]
  3. Fu S.-C.J., Birnbaum S. M., Greenstein J. P. 1954; Influence of optically active acyl compounds on the enzymatic hydrolysis of N-acylated-l-amino acids. Journal of the American Chemical Society 76:6054–6058
    [Google Scholar]
  4. Goldman P. 1965; The enzymatic cleavage of the C-F bond in fluoroacetate. Journal of Biological Chemistry 240:3434–3438
    [Google Scholar]
  5. Goldman P., Milne G. W. A., Keister D. B. 1968; Carbon-halogen bond cleavage. III. Studies on bacterial halidohydrolases. Journal of Biological Chemistry 243:428–434
    [Google Scholar]
  6. Hardman D. J., Slater J. H. 1981; Dehalogenases in soil bacteria. Journal of General Microbiology 123:117–128
    [Google Scholar]
  7. Hohorst H. J. 1965; l-(+)-Lactate.Determination with lactic dehydrogenase and DPN. In Methods of Enzymatic Analysis pp. 266–270 Bergmeyer H. U. Edited by New York:: Academic Press.;
    [Google Scholar]
  8. Kawasaki H., Tone N., Tonomura K. 1981a; Plasmid determined dehalogenation of haloacetates in Moraxella species. Agricultural and Biological Chemistry 45:29–34
    [Google Scholar]
  9. Kawasaki H., Tone N., Tonomura K. 1981b; Purification and properties of haloacetatehalidohydrolase specified by plasmid from Moraxella sp. strain B. Agricultural and Biological Chemistry 45:35–42
    [Google Scholar]
  10. Little M., Williams P. A. 1971; A bacterial halidohydrolase.Its purification, some properties and its modification by specific amino acid reagents. European Journal of Biochemistry 21:99–109
    [Google Scholar]
  11. Senior E., Bull A. T., Slater J. H. 1976; Enzyme evolution in a microbial community growing on the herbicide Dalapon. Nature; London: 263476–479
    [Google Scholar]
  12. Slater J. H., Lovatt D., Weightman A. J., Senior E., Bull A. T. 1979; The growth of Pseudomonas putida on chlorinated aliphatic acids and its dehalogenase activity. Journal of General Microbiology 114:125–136
    [Google Scholar]
  13. Weightman A. J., Slater J. H. 1980; Selection of Pseudomonas putida strains with elevated dehalogenase activities by continuous culture growth on chlorinated alkanoic acids. Journal of General Microbiology 121:187–193
    [Google Scholar]
  14. Weightman A. J., Slater J. H., Bull A. T. 1979; The partial purification of two dehalogenases from Pseudomonas putida PP3. FEMS Microbiology Letters 6:231–234
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-128-8-1755
Loading
/content/journal/micro/10.1099/00221287-128-8-1755
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error