Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Improvements of GC and HPLC Analyses in Solvent (Acetone-Butanol-Ethanol) Fermentation by Clostridium saccharobutylicum Using a Mixture of Starch and Glycerol as Carbon Source

  • Tsuey, Liew Shiau (Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia) ;
  • Ariff, Arbakariya Bin (Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia) ;
  • Mohamad, Rosfarizan (Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia) ;
  • Rahim, Raha Abdul (Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia)
  • Published : 2006.08.30
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Abstract

A study on the feasibility of using improved computer-controlled HPLC and GC systems was carried out to shorten the time needed for measuring levels of the substrates (glucose, maltose, and glycerol) and products (acetone, butanol ethanol, acetic acid, and butyric acid) produced by Clostridium saccharobutylicum DSM 13864 during direct fermentation of sago starch to solvent. The use of HPLC system with a single injection to analyse the composition of culture broth (substrates and products) during solvent fermentation was achieved by raising the column temperature to $80^{\circ}C$. Although good separation of the components in the mixture was achieved, a slight overlap was observed in the peaks for butyric acid and acetone. The shape of the peak obtained and the analysis time of 26.66 min were satisfactory at a fixed flow rate of 0.8mL/min. An improved GC system was developed, that was able to measure the products of solvent fermentation (acetone, butanol, ethanol, acetic acid, and butyric acid) within 19.28 min. Excellent resolution for each peak was achieved by adjusting the oven temperature to $65^{\circ}C$.

Keywords

References

  1. Ennis, B. M., N. A. Gutierrez, and I. S. Maddox (1986) The acetone-butanol-ethanol fermentation: A current assessment. Process Biochem. 21: 131-146
  2. Madihah, M. S., A. B. Ariff, M. S. Khalil, A. A. Suraini, and M. I. A. Karim (2001) Anaerobic fermentation of gelatinized sago starch-derived sugars to acetone-1-butanolethanol solvent by Clostridium acetobutylicum. Folia Microbiol. 46: 197-204 https://doi.org/10.1007/BF02818533
  3. Jones, D. T. and D. R. Woods (1986) Acetone-butanol fermentation revisited. Microbiol. Rev. 50: 484-524
  4. Mitchell, W. J. (2001) Biology and physiology. pp. 49-81. In: H. Bahl and P. Durre (eds.). Clostridia: Biotechnology and Medical Applications. Wiley-VCH Verlag GmbH, Weinhem, Germany
  5. Ezeji, T. C., N. Qureshi, and H. P. Blaschek (2004) Acetone butanol ethanol (ABE) production from concentrated substrate: reduction in substrate inhibition by fed-batch technique and product inhibition by gas stripping. Appl. Microbiol. Biotechnol. 63: 653-658 https://doi.org/10.1007/s00253-003-1400-x
  6. Ishizaki, A., S. Michiwaki, E. Crabbe, G. Kobayashi, K. Sonomoto, and S. Yoshino (1999) Extractive acetonebutanol- ethanol fermentation using methylated crude palm oil as extractant in batch culture of Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564). J. Biosci. Bioeng. 87: 352-356 https://doi.org/10.1016/S1389-1723(99)80001-2
  7. Qureshi, N., M. M. Meagher, J. Huang, and R. W. Hutkins (2001) Acetone butanol ethanol (ABE) recovery by pervaporation using silicate-silicone composite membrane from fed-batch reactor of Clostridium acetobutylicum. J. Membrane Sci. 187: 93-102 https://doi.org/10.1016/S0376-7388(00)00667-0
  8. Buday, Z., J. C. Linden, and M. N. Karim (1990) Improved acetone-butanol fermentation analysis using subambient HPLC column temperature. Enzyme Microb. Technol. 12: 24-27 https://doi.org/10.1016/0141-0229(90)90175-P
  9. Datta, R. and J. G. Zeikus (1985) Modulation of acetonebutanol- ethanol fermentation by carbon monoxide and organic acids. Appl. Environ. Microbiol. 49: 522-529
  10. Lopez-Contreras, A. M., P. A. M. Claassen, H. Mooibroek, and W. M. De Vos (2000) Utilisation of saccharides in extruded domestic organic waste by Clostridium acetobutylicum ATCC 824 for production of acetone, butanol and ethanol. Appl. Microbiol. Biotechnol. 54: 162-167 https://doi.org/10.1007/s002530000374
  11. Meyer, C. L. and E. T. Papoutsakis (1989) Increased levels of ATP and NADH are associated with increased solvent production in continuous cultures of Clostridium acetobutylicum. Appl. Microbiol. Biotechnol. 30: 450-459
  12. Saint-Amans, S., L. Girbal, J. Andrade, K. Ahrens, and P. Soucaille (2001) Regulation of carbon and electron flow in Clostridium butyricum VPI 3266 grown on glucose-glycerol mixtures. J. Bacteriol. 183: 1748-1754 https://doi.org/10.1128/JB.183.5.1748-1754.2001
  13. Pecina, R. and G. Bonn (1984) High-performance liquid chromatographic elution behaviour of alcohols, aldehydes, ketones, organic acids and carbohydrates on a strong cation-exchange stationary phase. J. Chromatogr. 287: 245-258 https://doi.org/10.1016/S0021-9673(01)87701-3
  14. Fowlis, I. A. (1995) Gas Chromatograph; Analytical Chemistry by Open Learning. 2nd ed., pp. 158-166. John Wiley and Sons, New York, NY, USA