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Applied Microbiology and Biotechnology

, Volume 79, Issue 2, pp 263–272 | Cite as

Development of chemically defined medium for Mannheimia succiniciproducens based on its genome sequence

  • Hyohak Song
  • Tae Yong Kim
  • Bo-Kyeong Choi
  • Seong Jun Choi
  • Lars K. Nielsen
  • Ho Nam Chang
  • Sang Yup LeeEmail author
Applied Microbial and Cell Physiology

Abstract

This study presents a novel methodology for the development of a chemically defined medium (CDM) using genome-scale metabolic network and flux balance analysis. The genome-based in silico analysis identified two amino acids and four vitamins as non-substitutable essential compounds to be supplemented to a minimal medium for the sustainable growth of Mannheimia succiniciproducens, while no substitutable essential compounds were identified. The in silico predictions were verified by cultivating the cells on a CDM containing the six non-substitutable essential compounds, and it was further demonstrated by observing no cell growth on the CDM lacking any one of the non-substitutable essentials. An optimal CDM for the enhancement of cell growth and succinic acid production, as a target product, was formulated with a single-addition technique. The fermentation on the optimal CDM increased the succinic acid productivity by 36%, the final succinic acid concentration by 17%, and the succinic acid yield on glucose by 15% compared to the cultivation using a complex medium. The optimal CDM also lowered the sum of the amounts of by-products (acetic, formic, and lactic acids) by 30%. The strategy reported in this paper should be generally applicable to the development of CDMs for other organisms, whose genome sequences are available.

Keywords

Chemically defined medium Mannheimia succiniciproducens Metabolic network Flux balance analysis Succinic acid 

Notes

Acknowledgments

This work was supported by the Genome-based Integrated Bioprocess Project of the Ministry of Science and Technology (No. 2005-01294) through the Korea Science and Engineering Foundation (KOSEF). Further supports by the LG Chem Chair Professorship and by the KOSEF through the Center for Ultramicrochemical Process Systems are appreciated.

References

  1. Hong SH, Kim JS, Lee SY, In YH, Choi SS, Rih J–K, Kim CH, Jeong H, Hur CG, Kim JJ (2004) The genome sequence of the capnophilic rumen bacterium Mannheimia succiniciproducens. Nature Biotechnol 22:1275–1281CrossRefGoogle Scholar
  2. Hungate RE (1966) The rumen and its microbes. Academic, New York, NYGoogle Scholar
  3. Jablonski PE, Jaworski M, Hovde CJ (1996) A minimal medium for growth of Pasteurella multocida. FEMS Microbiol Lett 140:165–169CrossRefGoogle Scholar
  4. Jang Y-S, Jung YR, Lee SY, Kim JM, Lee JW, Oh D-B, Kang HA, Kwon O, Jang SH, Song H, Lee SJ, Kang KY (2007) Construction and characterization of shuttle vectors for succinic acid producing rumen bacteria. Appl Environ Microbiol 73:5411–5420CrossRefGoogle Scholar
  5. Kim TY, Kim HU, Park JM, Song H, Kim JS, Lee SY (2007) Genome-scale analysis of Mannheimia succiniciproducens metabolism. Biotechnol Bioeng 97:657–671CrossRefGoogle Scholar
  6. Kim JM, Lee KH, Lee SY (2008) Development of a markerless gene knock-out system for Mannheimia succiniciproducens using a temperature-sensitive plasmid. FEMS Microbiol Lett 278:78–85CrossRefGoogle Scholar
  7. Lee Y, Lee SY (1996) Enhanced production of poly (3-hydroxybutyrate) by filamentation-suppressed recombinant Escherichia coli in a defined medium. J Environ Polymer Degrad 4:131–134CrossRefGoogle Scholar
  8. Lee PC, Lee SY, Hong SH, Chang HN (2002) Isolation and characterization of a new succinic acid-producing bacterium, Mannheimia succiniciproducens MBEL 55E, from bovine rumen. Appl Microbiol Biotechnol 58:663–668CrossRefGoogle Scholar
  9. Lee PC, Lee SY, Hong SH, Chang HN (2003a) Batch and continuous cultures of Mannheimia succiniciproducens MBEL55E for the production of succinic acid from whey and corn steep liquor. Bioproc Biosyst Eng 26:63–67CrossRefGoogle Scholar
  10. Lee DY, Yun HS, Lee SY, Park SW (2003b) MetaFluxNet: the management of metabolic reaction information and quantitative metabolic flux analysis. Bioinformatics 19:2144–2146CrossRefGoogle Scholar
  11. Lee JW, Lee SY, Song H, Yoo J-S (2006a) The proteome of Mannheimia succiniciproducens, a capnophilic rumen bacterium. Proteomics 6:3550–3566CrossRefGoogle Scholar
  12. Lee SJ, Song H, Lee SY (2006b) Genome-based metabolic engineering of Mannheimia succiniciproducens for succinic acid production. Appl Environ Microbiol 72:1939–1948CrossRefGoogle Scholar
  13. Lim H-K, Kim S-G, Jung K-H, Soo J-H (2004) Statistical selection of amino acids fortifying a minimal defined medium for a high-level production of the kringle fragments of human apolipoprotein. J Microbiol Biotechnol 14:90–96Google Scholar
  14. Mantha D, Basha ZA, Pnada T (1998) Optimization of medium composition by response surface methodology for the production of tartaric acid by Gluconobacter suboxydans. Bioproc Eng 19:285–288Google Scholar
  15. Marquis H, Bouwer HGA, Hinrichs DJ, Portnoy DA (1993) Intracytoplasmic growth and virulence of Listeria monocytogenes auxotrophic mutants. Infect Immun 61:3756–3760CrossRefGoogle Scholar
  16. McKinlay JB, Vieille C, Zeikus JG (2007) Prospects for bio-based succinate industry. Appl Microbiol Biotechnol 76:727–740CrossRefGoogle Scholar
  17. Nohata Y, Kurane R (1997) Complete defined medium for large-scale production of polysaccharide bioadsorbent from Alcaligenes latus B-16. J Ferment Bioeng 83:116–117CrossRefGoogle Scholar
  18. Preetha R, Jayaprakash NS, Philip R, Bright Singh IS (2007) Optimization of medium for the production of a novel aquaculture probiotic, Micrococcus MCCB 104 using central composite design. Biotechnol Bioprocess Eng 12:548–555CrossRefGoogle Scholar
  19. Premaratne RJ, Lin W-J, Johnson EA (1991) Development of an improved chemically defined minimal medium for Listeria monocytogenes. Appl Environ Microbiol 57:3046–3048CrossRefGoogle Scholar
  20. Saito K (2000) Regulation of sulfate transport and synthesis of sulphur-containing amino acids. Curr Opin Plant Biol 3:188–195CrossRefGoogle Scholar
  21. Scully LR, Bidochka MJ (2006) A cysteine/methionine auxotroph of the opportunistic fungus Aspergillus flavus is associated with host-range restriction: a model for emerging diseases. Microbiology 152:223–232CrossRefGoogle Scholar
  22. Shi F, Xu Z, Cen P (2006) Optimization of γ-polyglutamic acid production by Bacillus subtilis ZJU-7 using a surface-response methodology. Biotechnol Bioprocess Eng 11:251–257CrossRefGoogle Scholar
  23. Song H, Lee SY (2006) Production of succinic acid by bacterial fermentation. Enzyme Microb Technol 39:352–361CrossRefGoogle Scholar
  24. Song H, Lee JW, Choi S, You JK, Hong WH, Lee SY (2007) Effects of dissolved CO2 levels on the growth of Mannheimia succiniciproducens and succinic acid production. Biotechnol Bioeng 98:1296–1304CrossRefGoogle Scholar
  25. Song H, Jang SH, Park JM, Lee SY (2008) Modeling of batch fermentation kinetics for succinic acid production by Mannheimia succiniciproducens. Biochemical Eng J (in press). DOI  https://doi.org/10.1016/j.bej.2007.11.021 CrossRefGoogle Scholar
  26. Spaargaren DH (1996) The design of culture media based on the elemental composition of biological material. J Biotechnol 45:97–102CrossRefGoogle Scholar
  27. van der Werf MJ, Guettler MV, Jain MK, Zeikus JG (1997) Environmental and physiological factors affecting the succinate product ratio during carbohydrate fermentation by Actinobacillus sp. 130Z. Arch Microbiol 167:332–342CrossRefGoogle Scholar
  28. van Niel EWJ, Hahn-Hägerdal H (1999) Nutrient requirements of Lactococci in defined growth medium. Appl Microbiol Biotechnol 52:617–627CrossRefGoogle Scholar
  29. Varma A, Palsson BO (1994) Stoichiometric flux balance models quantitatively predict growth and metabolic by-product secretion in wild-type Escherichia coli W3110. Appl Environ Microbiol 60:3724–3731CrossRefGoogle Scholar
  30. Zang J, Greasham R (1999) Chemically defined medium for commercial fermentations. Appl Microbiol Biotechnol 51:407–421CrossRefGoogle Scholar
  31. Zeikus JG, Jain MK, Elankovan P (1999) Biotechnology of succinic acid production and markets for derived industrial products. Appl Microbiol Biotechnol 51:545–552CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Hyohak Song
    • 1
    • 2
    • 3
  • Tae Yong Kim
    • 1
    • 2
    • 3
  • Bo-Kyeong Choi
    • 1
    • 2
  • Seong Jun Choi
    • 1
    • 2
    • 3
  • Lars K. Nielsen
    • 5
  • Ho Nam Chang
    • 1
    • 2
  • Sang Yup Lee
    • 1
    • 2
    • 3
    • 4
    Email author
  1. 1.Department of Chemical and Biomolecular Engineering (BK21 Program)KAISTDaejeonRepublic of Korea
  2. 2.BioProcess Engineering Research CenterKAISTDaejeonRepublic of Korea
  3. 3.Metabolic and Biomolecular Engineering National Research Laboratory, Center for Systems and Synthetic Biotechnology, Institute for the BioCenturyKAISTDaejeonRepublic of Korea
  4. 4.Department of Bio and Brain Engineering and Bioinformatics Research CenterKAISTDaejeonRepublic of Korea
  5. 5.Australian Institute for Bioengineering and NanotechnologyUniversity of QueenslandBrisbaneAustralia

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