Abstract
Seven organophosphorus pesticide-degrading bacteria harboring the methyl parathion degrading (mpd) gene were isolated from a methyl parathion contaminated site. In this study, the 4.7 kb mpd gene cluster, conserved in all seven bacteria capable of degrading methyl parathion, was cloned and further analysis revealed that this cluster contained five ORFs and the mpd gene was associated with a mobile element, IS6100. In addition to mpd gene ORF and tnpA ORF, three other ORFs showed high homology to the permease component of ABC-type transport system, the general secretion pathway protein B, and the RNA polymerase sigma 70 factor, respectively. The mpd genes of these 7 strains were subcloned and expressed in E. coli, SDS-PAGE and zymogram analysis showed that two expression products of mpd genes in E. coli were found, but the one without signal peptide showed the hydrolytic activities. Our evidences collectively suggest that mpd gene cluster may be disseminated through horizontal gene transfer based on phylogenetic analysis of the cluster and their host bacterial strains, and comparisons of GC content of the cluster and respective host’s chromosome.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ, 1997 Gapped BLAST and PSI-BLAST: a new generation of protein database search programs Nucleic Acids Res. 25: 3389–3402
Chaudhry GR, Ali AN, Wheeler WB, 1988 Isolation of a methyl parathion-degrading Pseudomonas sp. that possesses DNA homologous to the opd gene from a Flavobacterium sp Appl. Environ. Microbiol. 54 288–293
Cho CM-H, Mulchandani A, Chen W, 2004 Altering the substrate specificity of organophosphorus hydrolase for enhanced hydrolysis of chlorpyrifos Appl. Environ. Microbiol. 70 4681–4685
Coenye T, Vancanneyt M, Falsen E, Swings J, Vandamme P, 2003, Achromobacter insolitus sp. nov. and Achromobacter spanius sp. nov., from human clinical samples Int. J. Syst. Bacteriol. 53 1819–1894
Cui ZL, Li SP, Fu GP, 2001 Isolation of methyl parathion-degrading strain M6 and cloning of the methyl parathion hydrolase gene Appl. Environ. Microbiol. 67 4922–4925
Fu GP, Cui ZL, Huang TT, Li SP, 2004 Expression, purification, and characterization of a novel methyl parathion hydrolase Protein Expres. Purif. 36 170–176
Granelli-Piperno A, Reich E, 1978 A study of proteases and protease-inhibitor complexes in biological fluids J. Exp. Med. 148 223–234
Hall TA, 1999 BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT Nucleic Acids Symp. Ser. 41 95–98
Halling SM, Peterson-Burch BD, Bricker BJ, Zuerner RL, Qing Z, Li LL, Kapur V, Alt DP, Olsen SC, 2005 Completion of the genome sequence of Brucella abortus and comparison to the highly similar genomes of Brucella melitensis and Brucella suis J. Bacteriol. 187 2715–2726
Horne I, Sutherland TD, Harcourt RL, Russell RJ, Oakeshott JG, 2002 Identification of an opd (organophosphate degradation) gene in an Agrobacterium isolate Appl. Environ. Microbiol. 68 3371–3376
Kampfer P, Muller C, Mau M, Neef A, Auling G, Busse HJ, Osborn AM, Stolz A, 1999, Description of Pseudaminobacter gen. nov. with two new species, Pseudaminobacter salicylatoxidans sp. nov. and Pseudaminobacter defluvii sp. nov Int. J. Syst. Bacteriol. 49 887–897
Li CZ, Zhang J, Jiang YP, Gurewich V, Chen Y, Liu JN, 2001 Urokinase-type plasminogen activator up-regulates. Its own expression by endothelial cells and monocytes via the u-PAR pathway Thrombosis Res. 103 221–232
Malone AS, Shellhammer TH, Courtney PD, 2002 Effects of high pressure on the viability, morphology, lysis, and cell wall hydrolase activity of Lactococcus lactis subsp. cremoris Appl. Environ. Microbiol. 68 4357–4363
McDaniel CS, Harper LL, Wild JR, 1988 Cloning and sequencing of a plasmid-borne gene (opd) encoding a phosphotriesterase J. Bacteriol. 170 2306–2311
Miller SA, Dykes DD, Polesky HF, 1988 A simple salting out procedure for extracting DNA from human nucleated cells Nucleic Acids Res. 16 1215
Mulbry WW, Karns JS, Kearney PC, Nelson JO, Mcdaniel CS, Wild JR, 1986 Identification of a plasmid-borne parathion hydrolase gene from Flavobacterium sp. by southern hybridization with opd from Pseudomonas diminuta Appl. Environ. Microbiol. 51 926–930
Mulbry WW, Kearney PC, Nelson JO, Karns JS, 1987 Physical comparison of parathion hydrolase plasmids from Pseudomonas diminuta and Flavobacterium sp Plasmid 18 173–177
Mulbry WW, Karns JS, (1989) Parathion hydrolase specified by the Flavobacterium opd gene-relationship between the gene and protein J. Bacteriol. 171 6740–6746
Nielsen H, Engelbrechet J, Brunak S, von Heijne G, 1997 Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites Protein Eng. 10 1–6
Serdar CM, Gibson DT, Munnecke DM, Lancaster JH, 1982 Plasmid involvement in parathion hydrolysis by Pseudomonas diminuta Appl. Environ. Microbiol. 44 246–249
Serdar CM, Murdock DC, Rohde MF, 1989 Parathion hydrolase gene from Pseudomonas diminuta MG: subcloning, complete nucleotide-sequence, and expression of the mature portion of the enzyme in Escherichia coli Bio/Technology 7 1151–1155
Sethunathan N, Yoshida T, 1973. A Flavobacterium sp. that degrades diazinon and parathion Can. J. Microbiol. 19 873–875
Siddavattam D, Khajamohiddin S, Manavathi B, Pakala SB, Merrick M, 2003 Transposon-like organization of the plasmid-borne organophosphate degradation (opd) gene cluster found in Flavobacterium sp Appl. Environ. Microbiol. 69 2533–2539
Somara S, Manavathi B, Tebbe C, Siddavattam D, 2002. Localisation of identical organophosphorus pesticide degrading (opd) genes on genetically dissimilar indigenous plasmids of soil bacteria: PCR amplification, cloning and sequencing of the opd gene from Flavobacterium balustinum Indian J. Exp. Biol. 40 774–779
Stormo GD, Schneider TD, Gold LM, 1982 Characterization of translational initiation sites in E. coli Nucleic Acids Res. 10 2971–2996
Tan HM, 1999 Bacterial catabolic transposons Appl. Microbiol. Biotechnol. 51 1–12
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG, 1997 The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools Nucleic Acids Res. 24 4876–4882
Vassalli JD, Dayer JM, Wohlwen A, 1984 Concomitant secretion of prourokinase and of a plasminogen activator inhibitor by cultured human monocytes macrophages J. Exp. Med. 159 1653–1658
Velasco J, Romero C, Lopez-Goni I, Leiva J, Diaz R, Moriyon I, 1998 Evaluation of the relatedness of Brucella spp. and Ochrobactrum anthropi and description of Ochrobactrum intermedium sp. nov., a new species with a closer relationship to Brucella spp Int. J. Syst. Bacteriol. 48 759–768
Wyndham RC, Cashore AE, Nakatsu CH, Peel MC, 1994 Catabolic transposons Biodegradation 5 323–342
Zhang RF, Dai QH, He J, Dai XZ, Li SP, 2004 Comparison of degrading characteristics of seven organophosphate pesticide-degrading bacteria Chin. Environ. Sci. 24 584–587
Acknowledgements
This work was supported by grants 30400014 and 680600016 from the Chinese National Natural Science Foundation, and by the Chinese National Programs for High Technology Research and Development (2003AA241150).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhang, R., Cui, Z., Zhang, X. et al. Cloning of the organophosphorus pesticide hydrolase gene clusters of seven degradative bacteria isolated from a methyl parathion contaminated site and evidence of their horizontal gene transfer. Biodegradation 17, 465–472 (2006). https://doi.org/10.1007/s10532-005-9018-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10532-005-9018-6