Journal of Microbiology and Biotechnology

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

Effects of Dissolved Oxygen Level on Avermectin $B_{1a}$ Production by Streptomyces avermitilis in Computer-Controlled Bioreactor Cultures

  • Song, Sung-Ki (Department of Molecular Bioscience, Kangwon National University) ;
  • Jeong, Yong-Seob (Faculty of Biotechnology, Chonbuk National University) ;
  • Kim, Pyeung-Hyeun (Department of Molecular Bioscience, Kangwon National University) ;
  • Chun, Gie-Taek (Department of Molecular Bioscience, Kangwon National University)
  • Published : 2006.11.30
Download PDF
⟨ Previous Next ⟩

Abstract

In order to investigate the effect of dissolved oxygen (DO) level on AVM $B_{1a}$ production by a high yielding mutant of Streptomyces avermitilis, five sets of bioreactor cultures were performed under variously controlled DO levels. Using an online computer control system, the agitation speed and aeration rate were automatically controlled in an adaptive manner, responding timely to the oxygen requirement of the producer microorganism. In the two cultures of DO limitation, the onset of AVM $B_{1a}$ biosynthesis was observed to casually coincide with the fermentation time when oxygen-limited conditions were overcome by the producing microorganism. In contrast, this phenomenon did not occur in the parallel fermentations with DO levels controlled at around 30% and 40% throughout the entire fermentation period, showing an almost growth-associated mode of AVM $B_{1a}$ production: AVM $B_{1a}$ biosynthesis under the environments of high DO levels started much earlier than the corresponding oxygen-limited cultures, leading to a significant enhancement of AVM $B_{1a}$ production during the exponential stage. Consequently, approximately 6-fold and 9-fold increases in the final AVM $B_{1a}$ production were obtained in 30% and 40% DO-controlled fermentations, respectively, especially when compared with the culture of severe DO limitation (the culture with 0% DO level during the exponential phase). The production yield ($Y_{p/x}$), volumetric production rate (Qp), and specific production rate (${\bar{q}}_p$) of the 40% DO-controlled culture were observed to be 14%, 15%, and 15% higher, respectively, than those of the parallel cultures that were performed under an excessive agitation speed (350 rpm) and aeration rate (1 vvm) to maintain sufficiently high DO levels throughout the entire fermentation period. These results suggest that high shear damage of the high-yielding strain due to an excessive agitation speed is the primary reason for the reduction of the AVM $B_{1a}$ biosynthetic capability of the producer. As for the cell growth, exponential growth patterns during the initial 3 days were observed in the fermentations of sufficient DO levels, whereas almost linear patterns of cell growth were observed in the other two cultures of DO limitation during the identical period, resulting in apparently lower amounts of DCW. These results led us to conclude that maintenance of optimum DO levels, but not too high to cause potential shear damage on the producer, was crucial not only for the cell growth, but also for the enhanced production of AVM $B_{1a}$ by the filamentous mycelial cells of Streptomyces avermitilis.

Keywords

References

  1. Brunker, P., W. Minas, P. T. Kallio, and J. E. Bailey. 1998. Genetic engineering of an industrial strain of Saccaropolyspora erythraea for stable expression of the Vitreoscilla haemoglobin gene (VHb). Microbiology 144: 2441-2448 https://doi.org/10.1099/00221287-144-9-2441
  2. Campbell, W. C., M. H. Fisher, E. O. Stapley, G. Albers-Schonberg, and T. A. Jacob. 1983. Ivermectin: A potent new antiparasitic agent. Science 221: 823-828 https://doi.org/10.1126/science.6308762
  3. Chen, S. T., O. D. Hensens, and M. D. Schulman. 1989. Biosynthesis, pp. 55-72. In W. C. Campbell (ed.), Ivermectin and Abamectin. Springer-Verlag, New York
  4. DeTilly, G., D. G. Mou, and C. L. Cooney. 1983. Optimization and economics of antibiotic production, pp. 190-209. In J. E. Smith, D. R. Berry, and B. Kristiansen (eds.), The Filamentous Fungi, vol. 4. Edward Arnold, London
  5. Dick, O., U. Onken, I. Sattler, and A. Zeeck. 1994. Influence of increased dissolved oxygen concentration on productivity and selectivity in cultures of a colabomycin-producing strain of Streptomyces griseoflavus. Appl. Microbiol. Biotechnol. 41: 373-377
  6. Doran, P. M. 1997. Bioprocess Engineering Principles, pp. 190-217. Academic Press, London
  7. Enfors, S. O. and B. Mattiasson. 1983. Oxygenation of processes involving immobilized cells, pp. 41-60. In B. Mattiasson (ed.), Immobilized Cells and Organelles, vol. 2. CRC Press, Boca Raton, FL
  8. Gbewonyo, K., D. Dimasi, and B. C. Buckland. 1987. Characterization of oxygen transfer and power absorption of hydrofoil impellers in viscous mycelial fermentations, pp. 128-234. In C. S. Ho and J. Y. Oldshue (eds.), Biotechnology Processes Scale-Up and Mixing. American Institute of Chemical Engineers, New York
  9. Hilgendorf, P., V. Heiser, H. Diekmann, and M. Thoma. 1987. Constant dissolved oxygen concentrations in cephalosporin C fermentation: Applicability of different controllers and effect on fermentation parameters. Appl. Microbiol. Biotechnol. 27: 247-251
  10. Ikeda, H., T. Nonomiya, and S. Omura. 2001. Organization of biosynthetic gene cluster for avermectin in Streptomyces avermitilis: Analysis of enzymatic domains in four polyketide synthases. J. Ind. Microbiol. Biotechnol. 27: 170-176 https://doi.org/10.1038/sj.jim.7000092
  11. Justen, P., G. C. Paul, A. W. Nienow, and C. R. Thomas. 1996. Dependence of mycelial morphology on impeller type and agitation intensity. Biotechnol. Bioeng. 52: 672-684 https://doi.org/10.1002/(SICI)1097-0290(19961220)52:6<672::AID-BIT5>3.0.CO;2-L
  12. Kaiser, D., U. Onken, I. Sattler, and A. Zeeck. 1994. Influence of increased dissolved oxygen concentration on the formation of secondary metabolites by manumycin-producing Streptomyces parvulus. Appl. Microbiol. Biotechnol. 41: 309-312 https://doi.org/10.1007/BF00221224
  13. Kim, C. Y., H. J. Park, Y. J. Yoon, H. Y. Kang, and E. S. Kim. 2004. Stimulation of actinorhodin production by Streptomyces lividans with a chromosomally-integrated antibiotic regulatory gene afsR2. J. Microbiol. Biotechnol. 14: 1089-1092
  14. Kohler, P. 2001. The biochemical basis of anthelmintic action and resistance. Int. J. Parasitol. 31: 336-345 https://doi.org/10.1016/S0020-7519(01)00131-X
  15. Magnolo, S. K., D. L. Leenutaphong, J. A. DeModena, J. E. Curtis, J. E. Bailey, J. L. Galazzo, and D. E. Hughes. 1991. Actinorhodin production by Streptomyces coelicolor and growth of Streptomyces lividans are improved by the expression of a bacterial hemoglobin. Biotechnology (NY) 9: 173-176 https://doi.org/10.1038/nbt0291-173
  16. Malik, V. S. 1980. Microbial secondary metabolism. Trends. Biochem. Sci. 5: 68-72 https://doi.org/10.1016/0968-0004(80)90071-7
  17. Park, H. S., S. H. Kang, H. J. Park, and E. S. Kim. 2005. Doxorubicin productivity improvement by the recombinant Streptomyces peucetius with high-copy regulatory genes cultured in the optimized media composition. J. Microbiol. Biotechnol. 15: 66-71
  18. Pfefferle, C., U. Theobald, H. Gurtler, and H. P. Fiedler. 2000. Improved secondary metabolite production in the genus Streptosporangium by optimization of the fermentation conditions. J. Biotechnol. 80: 135-142 https://doi.org/10.1016/S0168-1656(00)00249-2
  19. Robin, J., S. Bonneau, D. Schipper, H. Noorman, and J. Nielsen. 2003. Influence of the adipate and dissolved oxygen concentrations on the ${\beta}$-lactam production during continuous cultivations of a Penicillium chrysogenum strain expressing the expandase gene from Streptomyces clavuligerus. Metab. Eng. 5: 42-48 https://doi.org/10.1016/S1096-7176(03)00006-5
  20. Rollins, M. J., S. E. Jensen, and D. W. S. Westlake. 1988. Effect of aeration on antibiotic production by Streptomyces clavuligerus. J. Ind. Microbiol. 3: 357-364 https://doi.org/10.1007/BF01569557
  21. Rollins, M. J., S. E. Jensen, and D. W. S. Westlake. 1989. Regulation of antibiotic production by iron and oxygen during defined medium fermentations of Streptomyces clavuligerus. Appl. Microbiol. Biotechnol. 31: 390-396
  22. Roubos, J. A., P. Krabben, R. G. M. Luiten, H. B. Verbruggen, and J. J. Heijnen. 2001. A quantitative approach to characterizing cell lysis caused by mechanical agitation of Streptomyces clavuligerus. Biotechnol. Prog. 17: 336-347 https://doi.org/10.1021/bp0001617
  23. Song, S. K., Y. S. Jeong, and G. T. Chun. 2005. Development of avermectin $B_{1a}$ high-yielding mutants through rational screening strategy based on understanding of biosynthetic pathway of avermectin $B_{1a}$. Korean J. Biotechnol. Bioeng. 20: 471-477
  24. Steel, M. R. and W. E. Maxon. 1966. Dissolved oxygen measurements in pilot- and production-scale novobiocin fermentations. Biotechnol. Bioeng. 8: 97-108 https://doi.org/10.1002/bit.260080109
  25. Taguchi, H., T. Yoshida, Y. Tomita, and S. Teramoto. 1968. The effects of agitation on disruption of the mycelial pellets in stirred fermentors. J. Ferment. Technol. 10: 814-822
  26. van Sujidam, J. C. and B. Metz. 1981. Influence of engineering variables upon the morphology of filamentous molds. Biotechnol. Bioeng. 23: 111-148 https://doi.org/10.1002/bit.260230109
  27. Yegneswaran, P. K. and M. R. Gray. 1988. Effect of reduced oxygen on growth and antibiotic production in Streptomyces clavuligerus. Biotechnol. Lett. 10: 479-484 https://doi.org/10.1007/BF01027060
  28. Yoon, Y. J., E. S. Kim, Y. S. Hwang, and C. Y. Choi. 2004. Avermectin: Biochemical and molecular basis of its biosynthesis and regulation. Appl. Microbiol. Biotechnol. 63: 626-634 https://doi.org/10.1007/s00253-003-1491-4