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Physiological and fermentation properties of Bacillus coagulans and a mutant lacking fermentative lactate dehydrogenase activity

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Journal of Industrial Microbiology & Biotechnology

Abstract

Bacillus coagulans, a sporogenic lactic acid bacterium, grows optimally at 50–55°C and produces lactic acid as the primary fermentation product from both hexoses and pentoses. The amount of fungal cellulases required for simultaneous saccharification and fermentation (SSF) at 55°C was previously reported to be three to four times lower than for SSF at the optimum growth temperature for Saccharomyces cerevisiae of 35°C. An ethanologenic B. coagulans is expected to lower the cellulase loading and production cost of cellulosic ethanol due to SSF at 55°C. As a first step towards developing B. coagulans as an ethanologenic microbial biocatalyst, activity of the primary fermentation enzyme L-lactate dehydrogenase was removed by mutation (strain Suy27). Strain Suy27 produced ethanol as the main fermentation product from glucose during growth at pH 7.0 (0.33 g ethanol per g glucose fermented). Pyruvate dehydrogenase (PDH) and alcohol dehydrogenase (ADH) acting in series contributed to about 55% of the ethanol produced by this mutant while pyruvate formate lyase and ADH were responsible for the remainder. Due to the absence of PDH activity in B. coagulans during fermentative growth at pH 5.0, the l-ldh mutant failed to grow anaerobically at pH 5.0. Strain Suy27-13, a derivative of the l-ldh mutant strain Suy27, that produced PDH activity during anaerobic growth at pH 5.0 grew at this pH and also produced ethanol as the fermentation product (0.39 g per g glucose). These results show that construction of an ethanologenic B. coagulans requires optimal expression of PDH activity in addition to the removal of the LDH activity to support growth and ethanol production.

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Acknowledgments

We thank Dr. Julie A. Maupin-Furlow for the use of BioFlo fermenter. This study was supported in part by a grant from the Department of Energy (DE-FG36-04GO14019), the State of Florida, University of Florida Agricultural Experiment Station, and the Florida Energy Systems Consortium.

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Authors and Affiliations

  1. Department of Microbiology and Cell Science, University of Florida, Box 110700, Gainesville, FL, 32611, USA

    Yue Su, Mun Su Rhee, Lonnie O. Ingram & K. T. Shanmugam

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  1. Yue Su
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  2. Mun Su Rhee
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  3. Lonnie O. Ingram
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  4. K. T. Shanmugam
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Corresponding author

Correspondence to K. T. Shanmugam.

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Y. Su and M. S. Rhee contributed equally to this work.

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Su, Y., Rhee, M.S., Ingram, L.O. et al. Physiological and fermentation properties of Bacillus coagulans and a mutant lacking fermentative lactate dehydrogenase activity. J Ind Microbiol Biotechnol 38, 441–450 (2011). https://doi.org/10.1007/s10295-010-0788-4

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  • DOI: https://doi.org/10.1007/s10295-010-0788-4

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