Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Cloning of Four Genes Involved in Limonene Hydroxylation from Enterobacter cowanii 6L

  • Published : 2007.07.31
Download PDF
⟨ Previous Next ⟩

Abstract

Genes encoding proteins responsible for limonene catabolism were cloned from a limonene-degrading microorganism, Enterobacter cowanii 6L, which was isolated from citron (Citrus junos) peel. The 8.6, 4.7, and 7.7 kb fragments (CD3, CD4, and CD6) of E. cowanii 6L chromosomal DNA that confer to E. coli the ability to grow on limonene have been cloned and their corresponding DNA sequences were determined. Nine open reading frames (ORFs) were identified, and the four ORFs (921 bp of CD3-2; 1,515 bp of CD4-1; 1,776 bp of CD6-1; and 1,356 bp of CD6-2) that encode limonene hydroxylase were confirmed by independently expressing these genes in E. coli. FAD and NADH were found to stimulate the hydroxylation reaction if added to cell extracts from E. coli recombinants, and multiple compounds (linalool, dihydrolinalool, perillyl alcohol, (${\alpha}-terpineol$, and ${\gamma}-terpineol$) were the principal products observed. Our results suggest that the isolate E. cowanii 6L has a broad metabolic capability including utilization of limonene. This broad metabolic ability was confirmed by identifying four novel limonene hydroxylase functional ORFs in E. cowanii 6L.

Keywords

References

  1. Anandaraman, S. and G. A. Reineccius. 1986. Stability of encapsulate orange peel oil. Food Technol. 40: 88-93
  2. Baines, D. 2001. Analysis of purity, pp. 29. In Roe, S. (ed.), Protein Purification Techniques, 2nd Ed. Oxford University Press, New York, NY
  3. Barbirato, F., J. C. Verdoes, J. A. de Bont, and M. J. van der Werf. 1998. The Rhodococcus erythropolis DCL 14 limoneneI ,2-epoxide hydrolase gene encodes an enzyme belonging to a novel class of epoxide hydrolases. FEES Lett. 438: 293-296 https://doi.org/10.1016/S0014-5793(98)01322-2
  4. Bimboim, H. C. and J. Doly. 1979. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 7: 1513-1523 https://doi.org/10.1093/nar/7.6.1513
  5. Bowen, E. R. 1975. Potential by-products from the microbial transformation of D-limonene. Proc. Fla. State Hort. Soc. 88: 305-308
  6. Cadwallader, K. R., R. J. Braddock, M. E. Parish, and D. P. Higgins. 1989. Bioconversion of D-limonene by Pseudomonas gladioli. J Food Sci. 54: 1241-1245 https://doi.org/10.1111/j.1365-2621.1989.tb05964.x
  7. Chang, H. C. and P. J. Oriel. 1994. Bioproduction of perillyl alcohol and related monoterpenes by isolates of Bacillus stearothermophilus. J. Food Sci. 59: 660-662 https://doi.org/10.1111/j.1365-2621.1994.tb05588.x
  8. Chang, H. C., D. A. Gage, and P. J. Oriel. 1995. Cloning and expression of a Iimonene degradation pathway from Bacillus stearothermophilus in Escherichia coli. J. Food Sci. 60: 551-553 https://doi.org/10.1111/j.1365-2621.1995.tb09824.x
  9. Chatterjee, T. and D. K. Bhattacharyya. 2001. Biotransformation of limonene by Pseudomonas putida. Appl. Microbiol. Biotechnol. 55: 541-546 https://doi.org/10.1007/s002530000538
  10. Cheong, T. K. and P. J. Oriel. 2000. Cloning and expression of the Iimonene hydroxylase of Bacillus stearothermophilus BR388 and utilization in two-phase Iimonene conversions. Appl. Biochem. Biotechnol. 84: 903-915 https://doi.org/10.1385/ABAB:84-86:1-9:903
  11. Chin, H. S., F. Breidt, H. P. Fleming, W. Shin, and S. Yoon. 2006. Identifications of predominant bacterial isolates from the fermenting kimchi using ITS-PCR and partial 16S rDNA sequence analyses. J. Microbiol. Biotechnol. 16: 68-76
  12. Dhavalikar, R. S. and P. K. Bhattacharyya. 1966. Microbiological transformations of terpenes. Part VIII. Fermentation of Iimonene by a soil pseudomonad. Indian J. Biochem. 3: 144-157
  13. Dhavalikar, R. S., P. N. Rangachari, and P. K. Bhattacharyya. 1966. Microbiological transformations of terpenes. Part IX. Pathways of degradation of limonene in a soil pseudomonad. Indian J Biochem. 3: 158-164
  14. Duetz, W. A., A. H. M. FjaIlman, S. Ren, C. Jourdat, and B. Witholt. 2001. Biotransformation of D-limonene to (+) trans-carveol by toluene-grown Rhodococcus opacus PWD4 ceIls. Appl. Environ. Microbiol. 67: 2829-2832 https://doi.org/10.1128/AEM.67.6.2829-2832.2001
  15. Kim, G. Y. and M. H. Lee. 2006. Cloning and characterization of the urease gene cluster of Streptococcus vestibularis ATCC49124. J. Microbiol. Biotechnol. 16: 286-290
  16. Kim, T., H. Song, and H. Kim. 2005. Distribution of dominant Bifidobacteria in the intestinal microtlora of Korean adults and seniors, identified by SDS-PAGE of whole cell proteins and 16S rDNA sequence analysis. J. Microbiol. Biotechnol. 15: 388-394
  17. Kransnobajew, V. 1984. Terpenoids, Ch 4, pp. 98-125. In Lieslich, K. (ed.), Biotechnology-Biotransformations. Vol. 6a. Verlag Chemie, Weinheim
  18. Mars, A. E., J. P. L. Gorissen, I. van den Beld, and G. Eggink. 2001. Bioconversion of limonene to increased concentrations of perillic acid by Pseudomonas putida GS1in a fed-batch reactor. Appl. Microbiol. Biotechnol. 56: 101-107 https://doi.org/10.1007/s002530100625
  19. Park, Y. J., I. C. Kim, and H. C. Chang. 2003. Microbial conversion of (+)-limonene by an Enterobacter agglomerans isolate. J. Microbiol. Biotechnol. 13: 636-639
  20. Park, Y. J., I. C. Kim, H. H. Baek, O.-K. Bang, and H. C. Chang. 2004. Conversion of citron (Citrusjunos) peel oil by Enterobacter agglomerans. J. Microbiol. Biotechnol. 14: 1275-1279
  21. Quan, Z., S. Rhee, J. Bae, J. Baek, Y. Park, and S. Lee. 2006. Bacterial community structure in activated sludge reactors treating free or metal-complexed cyanides. J. Microbiol. Biotechnol. 16: 232-239
  22. Sadtler Research Laboratories. 1986. The Sadtler Standard Gas Chromatography Retention Index Library. Sadtler Laboratories, Philadelphia, U.S.A
  23. Saito, H. and K. I. Miura. 1963. Preparation of transforming deoxyribonucleic acid by phenol treatment. Biochim. Biophys. Acta 72: 619-629 https://doi.org/10.1016/0926-6550(63)90386-4
  24. Sambrook, J., E. F. Fritch, and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
  25. Uribe, S. and A. Pena. 1990. Toxicity of allelopathic monoterpene suspensions on yeast: Dependence on droplet size. J. Chem. Ecol. 16: 1399-1408 https://doi.org/10.1007/BF01021035
  26. van der Werf, M. J., P. M. Keijer, and P. H. van der Schaft. 2000. Xanthobacter sp. C20 contains a novel bioconversion pathway for limonene. J. Biotechnol. 84: 133-143 https://doi.org/10.1016/S0168-1656(00)00348-5
  27. Welsh, F. W., W. D. Murray, and R. E. Williams. 1989. Microbiological and enzymatic production of flavor and fragrance chemicals. Crit. Rev. Biotechnol. 9: 105-169 https://doi.org/10.3109/07388558909040617