Metabolic Flux Analysis of Beijerinckia indica for PS-7 Production

  • Wu Jian-Rong (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Son Jeong Hwa (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Seo Hyo-Jin (Interdisciplinary Program of Marine Biotechnology, Pukyong National University) ;
  • Kim Ki-Hong (Aquatic Life of Medicine, Pukyong National University) ;
  • Nam Yoon-Kwon (Aquaculture, Pukyong National University) ;
  • Lee Jin-Woo (Division of Biotechnology, Dong-A University) ;
  • Kim Sung-Koo (Department of Biotechnology and Bioengineering, Pukyong National University)
  • 발행 : 2005.02.01

초록

In order to investigate central metabolic changes in Beijerinckia indica, cells were grown on different carbon sources and intracellular flux distributions were studied under varying concentrations of nitrogen. Metabolic fluxes were estimated by combining material balances with extracellular substrate uptake rate, biomass formation rate, and exopolysaccharide (EPS) accumulation rate. Thirty-one metabolic reactions and 30 intracellular metabolites were considered for the flux analysis. The results revealed that most of the carbon source was directed into the Entner-Doudoroff pathway, followed by the recycling of triose-3-phosphate back to Hexose­6-phosphate. The pentose phosphate pathway was operated at a minimal level to supply the precursors for biomass formation. The different metabolic behaviors under varying nitrogen concentrations were observed with flux analysis.

키워드

참고문헌

  1. Kang, K. S. and W. H. McNeely (1976) Polysaccharide and bacterial fermentation process for its preparation. US Patent 3,960,832
  2. Lee, J. W., W. G. Yeomans, A. L. Allen, R. A. Gross, and D. L. Kaplan (1997) Compositional consistency of a heteropolysaccharide- 7 produced by Beijerinckia indica. Biotechnol. Lett. 19: 803-807 https://doi.org/10.1023/A:1018356713795
  3. Thorne, L., M. J. Mikolajczak, R. W. Armentrout, and T. J. Pollock (2000) Increasing the yield and viscosity of exopolysaccharide secreted by Sphingomonas by augmentation of chromosomal genes with multiple copies of cloned biosynthetic genes. J. Ind. Microbiol. Biotechnol. 25: 49-57 https://doi.org/10.1038/sj.jim.7000019
  4. Gulin, S., A. Kussak, P. E. Jansson, and G. Widmalm (2001) Structural studies of S-7, another exocellular polysaccharide containing 2-deoxy-arabino-hexuronic acid. Carbohyd. Res. 311: 285-290 https://doi.org/10.1016/S0008-6215(01)00050-7
  5. Standford, P. A. and J. Baird (1983) Industrial Utilization of Polysaccharide: In Polysaccharide II. Academic Press, London, UK
  6. Lee, S. Y. and E. T. Papoutsakis (1999) Metabolic Engineering. Marcel Dekker, NY, USA
  7. Cha, W. S., D. B. Choi, and S. H. Kang (2004) Optimization of culture meidia for solid-state culture of pleurotus feruae. Biotechnol. Bioprocess Eng. 9: 369-373 https://doi.org/10.1007/BF02933059
  8. Choi, A. S. G., Kim, B. D. Yoon, and H. M. Oh (2003) Growth and amino acid contents of spirulina platensis with different nitrogen sources. Biotechnol. Bioprocess Eng. 8: 368-374 https://doi.org/10.1007/BF02949281
  9. Dubois, M., K. A. Gilles, J. K. Hamilton, P. A. Rebers, and F. Smith (1956) Colorimetric method for determination of sugars and related substances. Anal. Chem. 28: 350-356 https://doi.org/10.1021/ac60111a017
  10. Koroleff, F. (1970) Direct Determination of Ammonia in Natural Waters as Indophenol Blue: In Information on Techniques and Methods for Seawater Analysis. pp. 19-22. Internat. Counc. Exploration of the sea, Charlottenlund, Norway
  11. Cataldo, D. A., M. Haroon, I. E. Scharader, and V. L. Youngs (1975) Rapid colorimetric determination of nitrate in plant tissue by nitration of salicylic acid. Commu. Soil Sci. Plan. 6: 71-80 https://doi.org/10.1080/00103627509366547
  12. Klinke, S., M. Dauner, G. Scott, B. Kessler, and B. Witholt (2000) Inactivation of isocitrate lyase leads to increased production of medium chain length poly (3-hydroxyalkanoates). Appl. Environ. Microbiol. 66: 929-913 https://doi.org/10.1128/AEM.66.3.909-913.2000
  13. Ketusse, F., P. Chevallereau, J. L. Simon, and N. Lindley (2002) The influence of metabolic network structures and energy requirements on xanthan gum yields. J. Biotechnol. 99: 307-317 https://doi.org/10.1016/S0168-1656(02)00221-3
  14. Vartak, B. N., C. C. Lin, J. M. Cleary, M. J. Fagan, and M. H. Jr. Saier (1995) Glucose metabolism in Sphingmonas elodea: pathway engineering via construction of a gucose- 6-phosphate dehydrogenase insertion mutant. Microbiology 141: 2339-2350 https://doi.org/10.1099/13500872-141-9-2339
  15. http://pseudocyc.pseudomonas.com:1555/
  16. http://genome.ornl.gov/microbial/avin/
  17. Neidhardt, F. C., J. L. Ingraham, and M. Schaechter (1990) Physiology of the Bacterial Cell: A Molecular Approach. Sinauer Associates Inc., Sunderland, Mass, UK
  18. Ampe, F., J. L. Uribelarrea, G. M. F. Aragao, and N. D. Lindley (1997) Benzoate degradation via the ortho pathway in Alcaligenes eutrophus is perturbed by succinate. Appl. Environ. Microbiol. 63: 2765-2770
  19. Valino, J. J. and G. Stephanopoulos (1993) Metabolic flux distribution in Corynebacterium glutmincum during growth and lysine overproduction. Biotechnol. Bioeng. 41: 633- 646 https://doi.org/10.1002/bit.260410606
  20. http://mbel.kaist.ac.kr/
  21. Schweizer, H. P. and P. Cecilia (1996) Regulation of glycerol metabolism in Pseudomonas aeruginosa: Characterization of the glpR repressor gene. J. Bacteriol. 178: 5215-5221 https://doi.org/10.1128/jb.178.17.5215-5221.1996
  22. ttp://www.genome.jp/kegg/pathway/
  23. Beale, J. M. Jr. and J. L. Foster (1996) Carbohydrate fluxes into alginate biosynthesis in Azotobacter vinelandii NCIB 8789: NMR investigations of the triose pools. Biochemistry 35: 4492-4501 https://doi.org/10.1021/bi951922v
  24. Portais, J. C., P. Tavernier, I. Gosselin, and J. N. Barbotin (1999) Cyclic organization of the carbohydrate metabolism in Sinohizobium meliloti. Eur. J. Biochem. 265: 473-480 https://doi.org/10.1046/j.1432-1327.1999.00778.x
  25. Gosselin, I., O. Wattraint, D. Riboul, J. N. Barbotin, and J. Charles (2001) A deep investigation on carbohydrate cycling in Sinohizobium meliloti. FEBS Lett. 499: 45-49 https://doi.org/10.1016/S0014-5793(01)02518-2