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Allopurinol-mediated lignocellulose-derived microbial inhibitor tolerance by Clostridium beijerinckii during acetone–butanol–ethanol (ABE) fermentation

  • Bioenergy and biofuels
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Abstract

In addition to glucans, xylans, and arabinans, lignocellulosic biomass hydrolysates contain significant levels of nonsugar components that are toxic to the microbes that are typically used to convert biomass to biofuels and chemicals. To enhance the tolerance of acetone–butanol–ethanol (ABE)-generating Clostridium beijerinckii NCIMB 8052 to these lignocellulose-derived microbial inhibitory compounds (LDMICs; e.g., furfural), we have been examining different metabolic perturbation strategies to increase the cellular reductant pools and thereby facilitate detoxification of LDMICs. As part of these efforts, we evaluated the effect of allopurinol, an inhibitor of NAD(P)H-generating xanthine dehydrogenase (XDH), on C. beijerinckii grown in furfural-supplemented medium and found that it unexpectedly increased the rate of detoxification of furfural by 1.4-fold and promoted growth, butanol, and ABE production by 1.2-, 2.5-, and 2-fold, respectively. Since NAD(P)H/NAD(P)+ levels in C. beijerinckii were largely unchanged upon allopurinol treatment, we postulated and validated a possible basis in DNA repair to account for the solventogenic gains with allopurinol. Following the observation that supplementation of allopurinol in the C. beijerinckii growth media mitigates the toxic effects of nalidixic acid, a DNA-damaging antibiotic, we found that allopurinol elicited 2.4- and 6.7-fold increase in the messenger RNA (mRNA) levels of xanthine and hypoxanthine phosphoribosyltransferases, key purine-salvage enzymes. Consistent with this finding, addition of inosine (a precursor of hypoxanthine) and xanthine led to 1.4- and 1.7-fold increase in butanol production in furfural-challenged cultures of C. beijerinckii. Taken together, our results provide a purine salvage-based rationale for the unanticipated effect of allopurinol in improving furfural tolerance of the ABE-fermenting C. beijerinckii.

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Acknowledgments

Salaries and research support were provided in part by State funds appropriated to the Ohio Plant Biotechnology Consortium by The Ohio State University, Ohio Agricultural Research and Development Center (OARDC), and the Hatch grant (Project No. OHO01333). We are grateful to Dr. Joseph Hogan (Department of Animal Sciences, The Ohio State University & OARDC) for allowing us use his iMark™ multi-well plate reader for NAD+/NADH and NADP+/NADPH assays.

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  1. Department of Animal Sciences, The Ohio State University, and Ohio State Agricultural Research and Development Center (OARDC), 305 Gerlaugh Hall, 1680 Madison Avenue, Wooster, OH, 44691, USA

    Victor Ujor, Chidozie Victor Agu & Thaddeus Chukwuemeka Ezeji

  2. Department of Chemistry and Biochemistry, and Center for RNA Biology, The Ohio State University, 484 West 12th Avenue, Columbus, OH, 43210, USA

    Venkat Gopalan

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  1. Victor Ujor
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Correspondence to Thaddeus Chukwuemeka Ezeji.

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Ujor, V., Agu, C.V., Gopalan, V. et al. Allopurinol-mediated lignocellulose-derived microbial inhibitor tolerance by Clostridium beijerinckii during acetone–butanol–ethanol (ABE) fermentation. Appl Microbiol Biotechnol 99, 3729–3740 (2015). https://doi.org/10.1007/s00253-015-6450-3

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