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Acetone production in solventogenic Clostridium species: new insights from non-enzymatic decarboxylation of acetoacetate

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Abstract

Development of a butanologenic strain with high selectivity for butanol production is often proposed as a possible route for improving the economics of biobutanol production by solventogenic Clostridium species. The acetoacetate decarboxylase (aadc) gene encoding acetoacetate decarboxylase (AADC), which catalyzes the decarboxylation of acetoacetate into acetone and CO2, was successfully disrupted by homologous recombination in solventogenic Clostridium beijerinckii NCIMB 8052 to generate an aadc − mutant. Our fermentation studies revealed that this mutant produces a maximum acetone concentration of 3 g/L (in P2 medium), a value comparable to that produced by wild-type C. beijerinckii 8052. Therefore, we postulated that AADC-catalyzed decarboxylation of acetoacetate is not the sole means for acetone generation. Our subsequent finding that non-enzymatic decarboxylation of acetoacetate in vitro, under conditions similar to in vivo acetone–butanol–ethanol (ABE) fermentation, produces 1.3 to 5.2 g/L acetone between pH 6.5 and 4 helps rationalize why various knock-out and knock-down strategies designed to disrupt aadc in solventogenic Clostridium species did not eliminate acetone production during ABE fermentation. Based on these results, we discuss alternatives to enhance selectivity for butanol production.

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Acknowledgments

This research was supported in part by the Ohio BioProducts Innovation Center (to TE), OARDC Seed Grant (to TE), Northeast Sun grant Initiative Award/Contract number GRT00012344 (to TE), and NSF grant MCB 0843543 (to VG).

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

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

    Bei Han & Thaddeus Chukwuemeka Ezeji

  2. Department of Biochemistry, The Ohio State University, 484 West 12th Avenue, Columbus, OH, 43210, USA

    Venkat Gopalan

  3. Department of Public Health, School of Medicine, Xi’an Jiaotong University, Xi’an, 710061, China

    Bei Han

Authors
  1. Bei Han
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  2. Venkat Gopalan
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  3. Thaddeus Chukwuemeka Ezeji
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Correspondence to Thaddeus Chukwuemeka Ezeji.

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Figure S1

Acetic and butyric acid production by C. beijerinckii NCIMB 8052 and C. beijerinckii 8052aadc− in P2 medium (a, c) and P2 medium containing 4g/L CaCO3 (b, d). Acid variations during growth of C. beijerinckii NCIMB 8052 and C. beijerinckii 8052aadc− indicate continuous production and re-assimilation of acetic and butyric acid during ABE fermentation (DOC 580 kb)

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Han, B., Gopalan, V. & Ezeji, T.C. Acetone production in solventogenic Clostridium species: new insights from non-enzymatic decarboxylation of acetoacetate. Appl Microbiol Biotechnol 91, 565–576 (2011). https://doi.org/10.1007/s00253-011-3276-5

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  • DOI: https://doi.org/10.1007/s00253-011-3276-5

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