1887

Abstract

Mini-Tn-induced mutants with defects in utilization of linear terpenes such as citronellol, geraniol, citronellate and/or geranylate were isolated from . One mutant was unable to utilize geraniol but showed wild-type growth with the three other acyclic terpenes tested. The Tn insertion site of the mutant was determined by DNA sequencing. Comparison with the genome sequence revealed that PA3028, an ORF with high similarity on the amino acid level to molybdenum cofactor biosynthesis protein A2 (encoded by ), was the target of mini-Tn in the mutant. Disruption of in PAO1 wild-type by insertion mutagenesis resulted in the same geraniol-minus phenotype. The ability to utilize geraniol was restored to the mutant by conjugative transfer of PCR-cloned wild-type on a broad-host-range plasmid. Growth of PAO1 on geraniol and geranial, but not on citronellol, citronellate or geranylate, was inhibited by the presence of 10 mM tungstate, a molybdenum-specific inhibitor. Inhibition by tungstate was prevented by addition of molybdate. The results indicate that at least one step in the oxidation of geraniol to geranic acid (geranial oxidation) is a molybdenum-dependent reaction in and is different from the molybdenum-independent oxidation of citronellol to citronellate.

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2005-07-01
2020-04-04
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References

  1. Cantwell S. G., Lau E. P., Watt D. S., Fall R. R. 1978; Biodegradation of acyclic isoprenoids by Pseudomonas species. J Bacteriol135:324–333
    [Google Scholar]
  2. de Lorenzo V., Herrero M., Jakubzik U., Timmis K. N. 1990; Mini-Tn 5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in Gram-negative eubacteria. J Bacteriol172:6568–6572
    [Google Scholar]
  3. Diaz-Perez A. L., Zavala-Hernandez A. N., Cervantes C., Campos-Garcia J. 2004; The gnyRDBHAL cluster is involved in acyclic isoprenoid degradation in Pseudomonas aeruginosa . Appl Environ Microbiol70:5102–5110[CrossRef]
    [Google Scholar]
  4. Fall R. R., Hector M. L. 1977; Acyl-coenzyme A carboxylases. Homologous 3-methylcrotonyl-CoA and geranyl-CoA carboxylases from Pseudomonas citronellolis . Biochemistry16:4000–4005[CrossRef]
    [Google Scholar]
  5. Hector M. L., Fall R. R. 1976; Multiple acyl-coenzymeA carboxylases in Pseudomonas citronellolis . Biochemistry15:3465–3472[CrossRef]
    [Google Scholar]
  6. Hille R. 1999; Molybdenum enzymes. Essays Biochem34:125–137
    [Google Scholar]
  7. Kovach M. E., Elzer P. H., Hill D. S., Robertson G. T., Farris M. A., Roop R. M., Peterson K. M. 1995; Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene166:175–176[CrossRef]
    [Google Scholar]
  8. Leimkühler S., Angermüller S., Schwarz G., Mendel R. R., Klipp W. 1999; Activity of the molybdopterin-containing xanthin dehydrogenase of Rhodobacter capsulatus can be restored by high molybdenum concentrations in a moeA mutant defective in molybdenum cofactor biosynthesis. J Bacteriol181:5930–5939
    [Google Scholar]
  9. Mergeay M., Nies D., Schlegel H. G., Gerits J., Charles P., Van Gijsegem F. 1985; Alcaligenes eutrophus CH34 is a facultative chemolithotroph with plasmid-bound resistance to heavy metals. J Bacteriol162:328–334
    [Google Scholar]
  10. Moura J. J., Brondino C. D., Trincao J., Romao M. J. 2004; Mo and W bis-MGD enzymes: nitrate reductases and formate dehydrogenases. J Biol Inorg Chem9:791–799[CrossRef]
    [Google Scholar]
  11. Nichols J., Rajagopalan K. V. 2002; Escherichia coli MoeA and MogA. Function in metal incorporation step of molybdenum cofactor biosynthesis. J Biol Chem277:24995–25000[CrossRef]
    [Google Scholar]
  12. Nichols J. D., Rajagopalan K. V. 2005; In vitro molybdenum ligation to molybdopterin using purified components. J Biol Chem280:7817–7822[CrossRef]
    [Google Scholar]
  13. Schlegel H. G., Kaltwasser H., Gottschalk G. 1961; A submersion method for culture of hydrogen-oxidizing bacteria: growth physiological studies. Arch Mikrobiol38:209–222[CrossRef]
    [Google Scholar]
  14. Seubert W. 1960; Degradation of isoprenoid compounds by microorganisms: I. Isolation and characterization of an isoprenoid-degrading bacterium, Pseudomonas citronellolis n.sp. J Bacteriol79:426–434
    [Google Scholar]
  15. Seubert W., Fass E. 1964a; Untersuchungen über den bakteriellen Abbau von Isoprenoiden: IV. Reinigung und Eigenschaften der β -Isohexenylglutaconyl-CoA-hydratase und β -Hydroxy- β -isohexenylglutaryl-CoA-lyase. Biochem Z341:23–34
    [Google Scholar]
  16. Seubert W., Fass E. 1964b; Untersuchungen über den bakteriellen Abbau von Isoprenoiden: V. Der Mechanismus des Isoprenoidabbaues. Biochem Z341:35–44
    [Google Scholar]
  17. Seubert W., Fass E., Remberger U. 1963; Untersuchungen über den bakteriellen Abbau von Isoprenoiden: III. Reinigung und Eigenschaften der Geranylcarboxylase. Biochem Z338:265–275
    [Google Scholar]
  18. Simon R., Priefer U., Pühler A. 1983; A broad host-range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Biotechnology1:784–791[CrossRef]
    [Google Scholar]
  19. Windgassen M., Urban A., Jaeger K. E. 2000; Rapid gene inactivation in Pseudomonas aeruginosa . FEMS Microbiol Lett193:201–205[CrossRef]
    [Google Scholar]
  20. Wuebbens M. M., Rajagopalan K. V. 2003; Mechanistic and mutational studies of Escherichia coli molybdopterin synthase clarify the final step of molybdopterin biosynthesis. J Biol Chem278:14523–14532[CrossRef]
    [Google Scholar]
  21. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene33:103–119[CrossRef]
    [Google Scholar]
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