Exponential Growth and Solvents-Production of Clostridium acetobutylicum ATCC 824 on TYA Media Containing Sucrose and Glucose as Different Sole Carbon Sources
American Journal of BioScience
Volume 5, Issue 4, July 2017, Pages: 64-69
Received: Mar. 26, 2017; Accepted: Apr. 19, 2017; Published: Jun. 19, 2017
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Authors
Elizabeth Omolola Oladapo, Department of Biological Sciences, Nigerian Defence Academy, Kaduna, Nigeria
Enimie Endurance Oaikhena, Department of Biological Sciences, Nigerian Defence Academy, Kaduna, Nigeria
Mohammed Sani Abdulsalami, Department of Biological Sciences, Nigerian Defence Academy, Kaduna, Nigeria
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Abstract
Clostridium acetobutylicum ATCC 824 is a solventogenic, obligate anaerobic bacterium that can grow on various types of carbohydrates and are capable of producing spores. In the present study, Clostridium acetobutylicum was successfully grown on TYA medium (tryptone, yeast, acetate medium) containing two different carbon sources, glucose and sucrose coupled with the production of acids (butyric and acetic acid) and solvents, ABE (acetone, butanol and ethanol). An investigation was undertaken to determine the impact of the two types of carbon sources on the solvent production and growth of Clostridium acetobutylicum. HPLC and GC analysis revealed the amount of acids and solvents produced respectively, as well as the amount of unutilized sugars. The amount of combined ABE produced on glucose (0.19g/l A, 0.39g/l B, 0.06g/l E) was higher than on sucrose as carbon source (0.15g/l A, 0.30g/l B, 0.03g/l E). The colony forming units of Clostridium acetobutylicum grown on glucose (4.70 x 105 units/ml) was higher than on sucrose (0.1 x 105) as judged by dilution spread plating on agar. Hence, Glucose was confirmed as the carbon source characterized by the best performance for solvents production and growth of the bacterium. The whole production process on both glucose and sucrose was observed to mainly influence the production of butanol with the concentration of 0.39g/l and 0.30g/l respectively, over the production of other solvents. Higher amount of solvents was produced at lower pH in both cultures with the different carbon sources. Wet-mounts, gram stain and endospore stain were used to determine the motility, type and sporulation of Clostridium acetobutylicum respectively. Acidogenic phase which seems to couple with the growth of vegetative cells, results into production of acetic and butyric acids. Solventogenic phase commences with a drop in pH and is accompanied by the onset of sporulation.
Keywords
Clostridium acetobutylicum ATCC 824, Sucrose, Glucose, ABE
To cite this article
Elizabeth Omolola Oladapo, Enimie Endurance Oaikhena, Mohammed Sani Abdulsalami, Exponential Growth and Solvents-Production of Clostridium acetobutylicum ATCC 824 on TYA Media Containing Sucrose and Glucose as Different Sole Carbon Sources, American Journal of BioScience. Vol. 5, No. 4, 2017, pp. 64-69. doi: 10.11648/j.ajbio.20170504.12
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Copyright © 2017 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
References
[1]
Bankar, S. B., Survase, S. A., Singhal, R. S. and Granstrom, T. (2011). Continuous two stage acetone-butanol-ethanol fermentation with integrated solvent removal using Clostridium acetobutylicum B 5313. Bioresource Technology. 106: 110-116.
[2]
Lynd, L. R., Weimer, P. J. and Zyl, W. H. (2002). Pretorius IS: Microbial cellulose utilization: fundamentals and biotechnology. Microbiology and Molecular Biology Reviews. 66(3): 506-577.
[3]
Barer, M. R. (2003). Physiological and molecular aspects of growth, non-growth, culturability and viability in bacteria. Cambridge University Press, Cambridge.
[4]
Li, J., Chen, J., Vidal, J. E. and McClane, B. A. (2011). The Agar-like quorum-sensing system regulates sporulation and production of enterotoxin and beta2 toxin by Clostridium perfringens type A non-food-borne human gastrointestinal disease strain F5603. Infection and Immunity. 79(6): 2451-2459.
[5]
Sandoval, N., Venkataramanan, K., Groth, T. and Papoutsakis, E. (2015). Whole-genome sequence of an evolved Clostridium pasteurianum strain reveals Spo0A deficiency responsible for increased butanol production and superior growth. Biotechnology for Biofuels. 8: 1.
[6]
Lutke-Eversloh, T. and Bahl, H. (2011). Metabolic engineering of Clostridium acetobutylicum: recent advances to improve butanol production. Current Opinion in Biotechnology. 22: 1-14.
[7]
Monot, F., Martin, J. R., Petitdemange, H. and Gay, R. (1982). Acetone and butanol production by Clostridium acetobutylicum in a synthetic medium. Applied and Environmental Microbiology. 44: 1318-1324.
[8]
Gehin, A., Gelhaye, E., Raval, G. and Petitdemange, H. (1995). Clostridium cellulolyticum Viability and Sporulation under Cellobiose Starvation Conditions. Applied and Environmental Microbiology. 61(3): 868-871.
[9]
Sarchami, E., Johnson, L. and Rehmann (2016). Optimization of fermentation condition favoring butanol production from glycerol by Clostridium pasteurianum DSM 525. Bioresource Technology 208: 73-80.
[10]
Servinsky, M. D., Kiel, J. T., Dupuy, N. F. and Sund, C. J. (2010). Transcriptomal analysis of differential carbohydrate utilization by Clostridium acetobutylicum. Microbiology. 156: 3478-3491.
[11]
Sreekumar, S., Baer, Z. C., Gross E. and Padmanaban (2014). S. chemo catalytic upgrading of tailored fermentation product toward biodiesel. Chem. Sus. Chem. 7: 2445 - 2448
[12]
Lowenthai, R. M. and Marsden, K. A. (1986). A rapid, simple method for Leukemia immunophenotyping using air-dried blood & bone marrow sensors. Journal of Immunological Methods. 93: 19-27.
[13]
Hucker, G. J. and Conn, H. J. (1923). Method of Gram Staining. New York State Agricultural Experiment Station Technical Bulletin. 93: 1-20.
[14]
Harley, J. P. and Prescott, L. M. (2002). Laboratory Exercises in Microbiology. 5th Edition, McGraw Hill, New York.
[15]
Gottschal, J. and Morris, J. C. (1981). Non-production of acetone and butanol by Clostridium acetobutylicum during glucose and ammonium-limitation in continuous culture. Biotechnology Letters. 3: 525-530.
[16]
Terracciano, J. S. and Kashket, E. R. (1986). Intracellular conditions required for initiation of solvent production by Clostridium acetobutylicum. Applied Environmental Microbiology. 52: 86–91.
[17]
Borman, S. (2014). Engineering Clostridium acetobutylicum for production of kerosene and biodiesel blendstock precursors. Metabolic Engineering. 25: 124-130.
[18]
Jones, D. T. and Woods, D. R. (1986). Acetone-butanol fermentation revisited. Microbiology Reviews. 50: 484-524.
[19]
Ehsaan, M., Kuit, W., Zhang, Y., Cartman, S., Heap, J., Winzer, K. and Minton, P. (2016). Mutant generation by allelic exchange and genome resequencing of the biobutanol organism Clostridium acetobutylicum ATCC 824. Biotechnology of Biofuels. 9: 4.
[20]
Mitchell, W. J. (1998). Physiology of carbohydrate to solvent conversion by clostridia. Advanced Microbial Physiology. 39: 31-130.
[21]
Kharkwal, S., Karimi, I. A., Chang, M. W. and Lee, D-Y (2009). Strain improvement and process development for biobutanol production. Recent Patents on Biotechnology. 3: 202-210.
[22]
Paredes, C. J., Alsaker, K. V. and Papoutsakis, E. T. (2005). A comparative genomic view of clostridial sporulation and physiology. Nature Reviews Microbiology. 3: 969-978.
[23]
Millat, T., Holger, J., Hubert, B., Ralf-Jorg, F. and Olaf, W. (2013). The pH-induced metabolic shift from acidogenesis to solventogenesis in Clostridium acetobutylicum. Experimental Standard Conditions of Enzyme Characterization. 1: 33-55.
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