Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1989 Dec;171(12):6740–6746. doi: 10.1128/jb.171.12.6740-6746.1989

Parathion hydrolase specified by the Flavobacterium opd gene: relationship between the gene and protein.

W W Mulbry 1, J S Karns 1
PMCID: PMC210571  PMID: 2556372

Abstract

The sequence of a 1,693-base-pair plasmid DNA fragment from Flavobacterium sp. strain ATCC 27551 containing the parathion hydrolase gene (opd) was determined. Within this sequence, there is only one open reading frame large enough to encode the 35,000-dalton membrane-associated hydrolase protein purified from Flavobacterium extracts. Amino-terminal sequence analysis of the purified Flavobacterium hydrolase demonstrated that serine is the amino-terminal residue of the hydrolase protein. The amino-terminal serine corresponds to a TCG codon located 87 base pairs downstream of the presumptive ATG initiation codon in the nucleotide sequence. The amino acid composition of the purified protein agrees well with that predicted from the nucleotide sequence, using serine as the amino-terminal residue. These data suggest that the parathion hydrolase protein is processed at its amino terminus in Flavobacterium sp. Construction in Escherichia coli of a lacZ-opd gene fusion in which the first 33 amino-terminal residues of opd were replaced by the first 5 residues of lacZ resulted in the production of an active hydrolase identical in molecular mass to the hydrolase isolated from Flavobacterium sp. E. coli cells containing the lacZ-opd fusion showed higher levels of hydrolase activity than did cells containing the parent plasmid.

Full text

PDF
6740

Images in this article

6744Image
on p.6744

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chaudhry G. R., Ali A. N., Wheeler W. B. Isolation of a methyl parathion-degrading Pseudomonas sp. that possesses DNA homologous to the opd gene from a Flavobacterium sp. Appl Environ Microbiol. 1988 Feb;54(2):288–293. doi: 10.1128/aem.54.2.288-293.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Deretic V., Chandrasekharappa S., Gill J. F., Chatterjee D. K., Chakrabarty A. M. A set of cassettes and improved vectors for genetic and biochemical characterization of Pseudomonas genes. Gene. 1987;57(1):61–72. doi: 10.1016/0378-1119(87)90177-6. [DOI] [PubMed] [Google Scholar]
  3. Dixon R. The xylABC promoter from the Pseudomonas putida TOL plasmid is activated by nitrogen regulatory genes in Escherichia coli. Mol Gen Genet. 1986 Apr;203(1):129–136. doi: 10.1007/BF00330393. [DOI] [PubMed] [Google Scholar]
  4. Harper L. L., McDaniel C. S., Miller C. E., Wild J. R. Dissimilar plasmids isolated from Pseudomonas diminuta MG and a Flavobacterium sp. (ATCC 27551) contain identical opd genes. Appl Environ Microbiol. 1988 Oct;54(10):2586–2589. doi: 10.1128/aem.54.10.2586-2589.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. McDaniel C. S., Harper L. L., Wild J. R. Cloning and sequencing of a plasmid-borne gene (opd) encoding a phosphotriesterase. J Bacteriol. 1988 May;170(5):2306–2311. doi: 10.1128/jb.170.5.2306-2311.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mulbry W. W., Karns J. S., Kearney P. C., Nelson J. O., McDaniel C. S., Wild J. R. Identification of a plasmid-borne parathion hydrolase gene from Flavobacterium sp. by southern hybridization with opd from Pseudomonas diminuta. Appl Environ Microbiol. 1986 May;51(5):926–930. doi: 10.1128/aem.51.5.926-930.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mulbry W. W., Karns J. S. Purification and characterization of three parathion hydrolases from gram-negative bacterial strains. Appl Environ Microbiol. 1989 Feb;55(2):289–293. doi: 10.1128/aem.55.2.289-293.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mulbry W. W., Kearney P. C., Nelson J. O., Karns J. S. Physical comparison of parathion hydrolase plasmids from Pseudomonas diminuta and Flavobacterium sp. Plasmid. 1987 Sep;18(2):173–177. doi: 10.1016/0147-619x(87)90046-1. [DOI] [PubMed] [Google Scholar]
  12. Munnecke D. M. Enzymatic hydrolysis of organophosphate insecticides, a possible pesticide disposal method. Appl Environ Microbiol. 1976 Jul;32(1):7–13. doi: 10.1128/aem.32.1.7-13.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sethunathan N., Yoshida T. A Flavobacterium sp. that degrades diazinon and parathion. Can J Microbiol. 1973 Jul;19(7):873–875. doi: 10.1139/m73-138. [DOI] [PubMed] [Google Scholar]
  15. Stormo G. D., Schneider T. D., Gold L. M. Characterization of translational initiation sites in E. coli. Nucleic Acids Res. 1982 May 11;10(9):2971–2996. doi: 10.1093/nar/10.9.2971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  17. von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. doi: 10.1111/j.1432-1033.1983.tb07424.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES