Abstract
Batch and semi-continuous thermophilic l-lactate fermentation experiments were performed using Bacillus coagulans and glucose as a substrate. Reactor performance and biomass concentrations were assessed using two methods: turbidity as a traditional biomass index and real-time polymerase chain reaction (PCR) quantification of 16S rRNA genes. In the batch experiment, although the relationship between turbidity and real-time PCR assay differed depending on the growth phase, a correlation was observed between both assay methods. In the semi-continuous experiment, real-time PCR measurement was well suited for use as an index for evaluating bacterial mass under different organic loading conditions. A mathematical model was applied to evaluate the real-time PCR quantification to long-term, semi-continuous lactate fermentation. Lactate fermentation was well suited since only B. coagulans was involved in the reactions. The results obtained revealed a fundamental relationship between real-time PCR and traditional biomass analyses.
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Batstone, D. J., J. Keller, I. Angelidaki, S. V. Kalyuzhnyi, S. G. Pavlostathis, A. Rozzi, W. T. M. Sanders, H. Siegrist, and V. A. Vavilin (2002) Anaerobic Digestion Model No.1. International Water Association (IWA) Publishing, London, UK.
Hidaka, T., T. Horie, S. Akao, and H. Tsuno (2010) Kinetic model of thermophilic L-lactate fermentation by Bacillus coagulans combined with real-time PCR quantification. Water Res. 44: 2554–2562.
Michelson, T., K. Kask, E. Jõgi, E. Talpsep, I. Suitso, and A. Nurk (2006) L(+)-Lactic acid producer Bacillus coagulans SIM-7 DSM 14043 and its comparison with Lactobacillus delbrueckii ssp. lactis DSM 20073. Enz. Microbial. Technol. 39: 861–867.
Payot, T., Z. Chemaly, and M. Fick (1999) Lactic acid production by Bacillus coagulans-kinetic studies and optimization of culture medium for batch and continuous fermentations. Enz. Microbial. Technol. 24: 191–199.
Kobayashi, T., D. Yasuda, Y. Li, K. Kubota, H. Harada, and H. Yu (2009) Characterization of start-up performance and archaeal community shifts during anaerobic self-degradation of waste-activated sludge. Bioresour. Technol. 100: 4981–4988.
Furet, J., P. Quénée, and P. Tailliez (2004) Molecular quantification of lactic acid bacteria in fermented milk products using real-time quantitative PCR. Int. J. Food Microbiol. 97: 197–207.
Wang, M. Y., B. H. Olson, and J. S. Chang (2008) Relationship among growth parameters for Clostridium butyricum, hydA gene expression, and biohydrogen production in a sucrose-supplemented batch reactor. Appl. Microbiol. Biotechnol. 78: 525–532.
Wang, M., Y. Tsai, B. H. Olson, and J. Chang (2008) Monitoring dark hydrogen fermentation performance of indigenous Clostridium butyricum by hydrogenase gene expression using RT-PCR and qPCR. Int. J. Hydrogen Energy 33: 4730–4738.
Akao, S., H. Tsuno, T. Horie, and S. Mori (2007) Effects of pH and temperature on products and bacterial community in L-lactate batch fermentation of garbage under unsterile condition. Water Res. 41: 2636–2642.
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.
APHA, AWWA, WEF (1995) Standard methods for the examination of water and wastewater. 19th ed., Washington D. C., American Public Health Association Publications.
Lin, J. Q., S. M. Lee, and Y. M. Koo (2004) Modeling and simulation of lactic acid fermentation with inhibition effects of lactic acid and glucose. Biotechnol. Bioproc. Eng. 9: 52–58.
Haldane, J. B. S. (1930) Enzymes. Longmans, London
Burgos-Rubio, C. N., M. R. Okos, and P. C. Wankat (2000) Kinetic study of the conversion of different substrates to lactic acid using Lactobacillus bulgaricus. Biotechnol. Prog. 16: 305–314.
Gray, N. F. (2004) Biology of Wastewater Treatment. 2nd ed., Imperial College Press, London, UK.
Cicek, N., J. Macomber, J. Davel, M. T. Suidan, J. Audic, and P. Genestet (2001) Effect of solids retention time on the performance and biological characteristics of a membrane bioreactor. Water Sci. Technol. 43: 43–50.
Tsuno, H., T. Hidaka, and F. Nishimura (2002) A simple biofilm model of bacterial competition for attached surface. Water Res. 36: 996–1006.
Hidaka, T., H. Tsuno, and N. Kishimoto (2003) Advanced treatment of sewage by pre-coagulation and biological filtration process. Water Res. 37: 4259–4269.
Angelidaki, I., M. Alves, D. Bolzonella, L. Borzacconi, J. L. Campos, A. J. Guwy, S. Kalyuzhnyi, P. Jenicek, and J. B. van Lier (2009) Defining the biomethane potential (BMP) of solid organic wastes and energy crops: A proposed protocol for batch assays. Water Sci. Technol. 59: 927–934.
Harms, G., A. C. Layton, H. M. Dionisi, I. R. Gregory, V. M. Garrett, S. A. Hawkins, K. G. Robinson, and G. S. Sayler (2003) Real-time pcr quantification of nitrifying bacteria in a municipal wastewater treatment plant. Environ. Sci. Technol. 37: 343–351.
Ha, J. H., T. Hidaka, and H. Tsuno (2011) Analysis of factors affecting the ratio of microcystin to chlorophyll-a in cyanobacterial blooms using real-time polymerase chain reaction. Environ. Toxicol. 26: 21–28.
Hidaka, T., T. Asahira, H. Koshikawa, J. Cheon, Y. Park, and H. Tsuno (2010) Effect of microbial composition on thermophilic acid fermentation. Enz. Microb. Technol. 47: 127–133.
Ramirez, I., E. I. P. Volcke, R. Rajinikanth, and J. Steyer (2009) Modeling microbial diversity in anaerobic digestion through an extended ADM1 model. Water Res. 43: 2787–2800.
Cheon, J., T. Hidaka, S. Mori, H. Koshikawa, and H. Tsuno (2008) Applicability of random cloning method to analyze microbial community in full-scale anaerobic digesters. J. Biosci. Bioeng. 106: 134–140.
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Hidaka, T., Tsuno, H., Yagi, H. et al. Fundamental analysis of real-time PCR quantification and modeling for thermophilic L-lactate fermentation by Bacillus coagulans from glucose. Biotechnol Bioproc E 17, 290–297 (2012). https://doi.org/10.1007/s12257-011-0370-6
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DOI: https://doi.org/10.1007/s12257-011-0370-6