1887

Abstract

SUMMARY: Using a DNA probe from the gene encoding squalene-hopene cyclase (SHC, EC 5.4.99.−) from the Gram-positive bacterium Alicyclobacillus acidocaldarius, we have cloned a 4·3 kb Hindll fragment of chromosomal DNA from Zymomonas mobilis. An open reading frame of 1977 bp was detected that could encode a protein of 658 amino acids with a calculated molecular mass of 74077 Da. Under the control of lac or tac promoters, this gene, shc, was expressed in Escherichia coli K12 strains and its product had squalene-hopene cyclase activity. Sequence alignments with the A. acidocaldarius SHC, the lanosterol cyclase of the yeast Candida albicans, and the cycloartenol synthase of the plant Arabidopsis thaliana revealed six highly conserved regions (mainly in the C-terminal part) of the proteins. These regions contained the core motif Gln-X-X-X-Gly-X-Trp.

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1995-01-01
2021-05-17
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References

  1. Abe I., Rohmer M., Prestwich G. D. 1993; Enzymatic cyclization of squalene and oxidosqualene to sterols and triterpenes. Chem Rev 93:2189–2206
    [Google Scholar]
  2. Armstrong G. A., Hundle B. S., Hearst J. E. 1993; Evolutionary conservation and structural similarities of carotenoid biosynthesis gene products from photosynthetic and non-photosynthetic organisms. Methods Enzymol 214:297–311
    [Google Scholar]
  3. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
    [Google Scholar]
  4. Bringer S., Härtner T., Poralla K., Sahm H. 1985; Influence of ethanol on the hopanoid content and the fatty acid pattern in batch and continuous cultures of Zymomonas mobilis . Arch Microbiol 140:312–316
    [Google Scholar]
  5. Buntel C. J., Griffin J. H. 1992; Nucleotide and deduced amino acid sequences of the oxidosqualene cyclase from Candida albicans . J Am Chem Soc 114:9711–9713
    [Google Scholar]
  6. Corey E. J., Matsuda S. P. T., Bartel B. 1993; Isolation of an Arabidopsis thaliana gene encoding cycloartenol synthase by functional expression in a yeast mutant lacking lanosterol synthase by the use of a chromatographic screen . Proc Natl Acad Set USA 90:11628–11632
    [Google Scholar]
  7. Eddy C. K., Smith O. H., Noel K. D. 1988; Cosmid cloning of five Zymomonas trp genes by complementation of Escherichia coli and Pseudomonas putida trp mutants . J Bacteriol 170:3158–3163
    [Google Scholar]
  8. Gold L., Pribnow D., Schneider T., Shinedling S., Singer B. S., Stormo G. 1981; Translation initiation in prokaryotes. Annu Rev Microbiol 45:747–803
    [Google Scholar]
  9. Goncalves de Lima O., De Araujo J. M., Schumacher I. F., Cavalcanti da Silva E. 1970; Estudos de microorganismos antagonistas presentes nas bebidas fermentadas usadas pelo povo do Recife. I Sobre uma variedade deZymomonas mobilis (Lindner) 1928 . Rev Instit Antibiot Univ Recife 10:3–15
    [Google Scholar]
  10. Gough J. A., Murray N. E. 1983; Sequence diversity among related genes for recognition of specific targets in DNA molecules. J Mol Biol 166:1–19
    [Google Scholar]
  11. Henikoff S. 1984; Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359
    [Google Scholar]
  12. Hermans M. A. F., Neuss B., Sahm H. 1991; Content and composition of hopanoids in Zymomonas mobilis under various growth conditions . J Bacteriol 173:5592–5595
    [Google Scholar]
  13. Horbach S., Neuss B., Sahm H. 1991; Effect of azasqualene on hopanoid biosynthesis and ethanol tolerance of Zymomonas mobilis . FEMS Microbiol Lett 79:347–350
    [Google Scholar]
  14. Ingram L. O. 1986; Microbial tolerance to alcohols: role of the cell membrane. Trends Biotechnol 4:40–44
    [Google Scholar]
  15. Kessler C., Höltke H. J., Seibl R., Burg J., Mühlegger K. 1990; Non-radioactive labeling and detection of nucleic acids. Biol Chem Hoppe-Seyler 371:917–927
    [Google Scholar]
  16. Low B. 1968; Formation of merodiploids with a class of rec- recipient strains of Escherichia coli K12 . Proc Natl Acad Sci USA 60:160–167
    [Google Scholar]
  17. Mullis K. B., Faloona F. A. 1987; Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol 155:335–350
    [Google Scholar]
  18. Neumann S., Simon H. 1986; Purification, partial charac terization and substrate specificity of a squalene cyclase from Bacillus acidocaldarius . Biol Chem Hoppe-Seyler 367:723–729
    [Google Scholar]
  19. Ochs D., Tappe C. H., Gärtner P., Kellner R., Poralla K. 1990; Properties of purified squalene-hopene cyclase from Bacillus acidocaldarius . Eur J Biochem 194:75–80
    [Google Scholar]
  20. Ochs D., Kaletta C., Entian K.-D., Beck-Sickinger A., Poralla K. 1992; Cloning, expression, and sequencing of squalene-hopene cyclase, a key enzyme in triterpenoid metabolism. J Bacteriol 174:298–302
    [Google Scholar]
  21. Ourisson G., Albrecht P., Rohmer M. 1984; The microbial origin of fossil fuels. Sci Am 245:34–41
    [Google Scholar]
  22. Poralla K., Kannenberg E. 1987; Hopanoids: sterol equivalents in bacteria. Am Chem Soc Symp Ser 325:239–251
    [Google Scholar]
  23. Poralla K., Hewelt A., Prestwich G. D., Abe I., Reipen I., Sprenger G. A. 1994; A specific amino acid repeat in squalene and oxidosqualene cyclases. Trends Biochem Sci 19:157–158
    [Google Scholar]
  24. Queen C., Korn L. J. 1984; A comprehensive sequence analysis program for the IBM personal computer. Nucleic Acids Res 12:581–599
    [Google Scholar]
  25. Roessner C. A., Min C., Hardin S. H., Harris-Haller L. W., McCollum J. C., Scott A. I. 1993; Sequence of the Candida albicans erg 7 gene . Gene 127:149–150
    [Google Scholar]
  26. Rohmer M., Anding C., Ourisson G. 1980; Non-specific biosynthesis of hopane triterpenes by a cell-free system from Acetobacter pasteurianum . Eur J Biochem 112:541–547
    [Google Scholar]
  27. Sahm H., Rohmer M., Bringer-Meyer S., Sprenger G. A., Welle R. 1993; Biochemistry and physiology of hopanoids in bacteria. Adv Microb Physiol 35:247–273
    [Google Scholar]
  28. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
    [Google Scholar]
  30. Schulenberg-Schell H., Neuss B., Sahm H. 1989; Quantitative determination of various hopanoids in microorganisms. Anal Biochem 181:120–124
    [Google Scholar]
  31. Seckler B., Poralla K. 1986; Characterization and partial purification of squalene-hopene-cyclase from Bacillus acidocaldarius . Biochim Biophys Acta 881:356–363
    [Google Scholar]
  32. Southern E. M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mot Biol 98:503–517
    [Google Scholar]
  33. Tabor S., Richardson C. C. 1987; DNA sequence analysis with a modified bacteriophage T7 polymerase. Proc Natl Acad Sci USA 84:4767–4771
    [Google Scholar]
  34. Tappe C. H. 1993; Squalen-Hopen-Cyclasen: Reinigung, Charakterisierung und Inhibitor-Experimente. PhD thesis University of Tubingen; Germany:
    [Google Scholar]
  35. Taylor R. F. 1984; Bacterial triterpenoids. Microbiol Rev 48:181–198
    [Google Scholar]
  36. Tinoco I., Borer P. N., Dengeler B., Levine M. J., Uhlenbeck C. O., Crothers D. M., Gralla J. 1973; Improved estimation of secondary structure in ribonucleic acids. Nature New Biol 246:40–41
    [Google Scholar]
  37. Uhlenbusch I., Sahm H., Sprenger G. A. 1991; Expression of an L-alanine dehydrogenase gene in Zymomonas mobilis and excretion of L-alanine . Appl Environ Microbiol 57:1360–1366
    [Google Scholar]
  38. Vieira J., Messing J. 1982; The pUC plasmids, an Ml3 mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
    [Google Scholar]
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