1887

Abstract

A PCR method suitable for the isolation of lipase genes directly from environmental DNA is described. The problems associated with the low levels of similarity between lipase genes were overcome by extensive analysis of conserved regions and careful primer design. Using this method, a lipase gene (-lipase) was isolated directly from environmental DNA. This lipase showed less than 20% similarity with other known lipases at the amino acid level. The study also revealed that distantly related members of the α/β hydrolase superfamily share similar conserved motifs with the lipases, thus making these genes targets for gene prospecting by PCR.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-148-8-2283
2002-08-01
2024-05-02
Loading full text...

Full text loading...

/deliver/fulltext/micro/148/8/1482283a.html?itemId=/content/journal/micro/10.1099/00221287-148-8-2283&mimeType=html&fmt=ahah

References

  1. Bell P. J. L., Nevalainen H., Morgan H. W., Bergquist P. L. 1999; Rapid cloning of thermoalkalaophilic lipases from Bacillus spp. Biotechnol Lett 21:1003–1006 [CrossRef]
    [Google Scholar]
  2. Britton H. T. S., Robinson R. A. 1931; Universal buffer solutions and the dissociation constant of veronal. J Chem Soc 19311456–1462
    [Google Scholar]
  3. Chung G. H., Lee Y. P., Jeohn G. H., Yoo O. J., Rhee J. S. 1991; Cloning and nucleotide sequence of thermostable lipase gene from Pseudomonas fluorescens . Agric Biol Chem 55:2359–2365 [CrossRef]
    [Google Scholar]
  4. Cousin X., Hotelier T., Giles K., Lievin P., Toutant J. P., Chatonnet A. 1997; The alpha/beta fold family of proteins database and the cholinesterase gene server. Nucleic Acids Res 25:143–146 [CrossRef]
    [Google Scholar]
  5. Dartois V. A., Baulard K., Schanck K., Colson C. 1992; Cloning, nucleotide sequence and expression in Escherichia coli of a lipase gene from Bacillus subtilis 168. Biochim Biophys Acta 1131:253–260 [CrossRef]
    [Google Scholar]
  6. Ferrato F., Carrière F., Sarda L., Verger R. 1997; A critical re-evaluation of the phenomenom of ‘interfacial activation’. Methods Enzymol 286:327–347
    [Google Scholar]
  7. Haas M. J., Allen J., Berka T. R. 1991; Cloning, expression and characterization of a cDNA encoding a lipase from Rhizopus delemar . Gene 109:107–113 [CrossRef]
    [Google Scholar]
  8. Ishida M., Oshima T. 1996; A leader open reading frame is essential for the expression in Escherichia coli of GC-rich leuB gene of an extreme thermophile, Thermus thermophilus. FEMS. Microbiol Rev 135:137–142 [CrossRef]
    [Google Scholar]
  9. Jaeger K.-E., Ransac S., Dijkstra B. W., Colson C., van Heuvel M., Misset O. 1994; Bacterial lipases. FEMS Microbiol Rev 15:29–63 [CrossRef]
    [Google Scholar]
  10. Jaeger K.-E., Dijkstra B. W., Reetz M. T. 1999; Bacterial biocatalysts: Molecular biology, three dimensional structures, and biotechnological applications of lipases. Annu Rev Microbiol 53:315–351 [CrossRef]
    [Google Scholar]
  11. Kim H.-Y., Park S.-Y., Lee J.-K., Ho T.-K. 1998; Gene cloning and characterization of thermostable lipase from Bacillus stearothermophilus L1. Biosci Biotechnol Biochem 65:66–71
    [Google Scholar]
  12. Kouker G., Jaeger K.-E. 1987; Specific and sensitive plate assay for bacterial lipases. Appl Environ Microbiol 53:211–213
    [Google Scholar]
  13. Lawrence R. C., Fryer T. F., Reiter B. 1967; Rapid method for the quantitative estimation of microbial lipases. Nature 213:589–597 [CrossRef]
    [Google Scholar]
  14. Morris D. D., Reeves R. A., Gibbs M. D., Saul D. J., Bergquist P. L. 1995; Correction of the β-mannanase domain of the celC pseudogene from Caldicellulosiruptor saccharolyticus and activity of the gene product on kraft pulp. Appl Environ Microbiol 61:2262–2269
    [Google Scholar]
  15. Morris D. D., Gibbs M. D., Chin C. W., Koh M. H., Wong R. W., Allison R. W., Nelson P. J., Bergquist P. L. 1998; Cloning of the xynB gene from Dictyoglomus thermophilum Rt46B.1 and action of the gene product on kraft pulp. Appl Environ Microbiol 64:1759–1765
    [Google Scholar]
  16. Oh B.-C., Kim H.-K., Lee J.-K., Kang S.-C., Oh T.-K. 1999; Staphylococcus haemolyticus lipase: biochemical properties, substrate specificity and gene cloning. FEMS Microbiol Lett 179:385–392 [CrossRef]
    [Google Scholar]
  17. Rose T. M., Schultz E. R., Henikoff J. G., Pietrokovski S., Henikoff S. 1998; Consensus-degenerate hybrid oligonucleotide primers for amplification of distantly related sequences. Nucleic Acids Res 26:1628–1635 [CrossRef]
    [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467 [CrossRef]
    [Google Scholar]
  19. Simons J. W., van Kampen M. D., Riel S., Gotz F., Egmond M. R., Verheij H. M. 1998; Cloning, purification and characterization of the lipase from Staphylococcus epidermidis – comparison of the substrate selectivity with those of other microbial lipases. Eur J Biochem 253:675–683 [CrossRef]
    [Google Scholar]
  20. Sullivan E. R., Leahy J. G., Colwell R. R. 1999; Cloning and sequence analysis of the lipase and lipase chaperone-encoding genes from Acinetobacter calcoaceticus RAG-1, and redefinition of a proteobacterial lipase family and an analogous lipase chaperone family. Gene 230:277–285 [CrossRef]
    [Google Scholar]
  21. van Pouderoyen G., Eggert T., Jaeger K. E., Dijkstra B. W. 2001; The crystal structure of Bacillus subtilis lipase: a minimal alpha/beta hydrolase fold enzyme. J Mol Biol 309:215–226 [CrossRef]
    [Google Scholar]
  22. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-148-8-2283
Loading
/content/journal/micro/10.1099/00221287-148-8-2283
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