Enhanced direct fermentation of cassava to butanol by Clostridium species strain BOH3 in cofactor-mediated mediumReport as inadecuate

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Biotechnology for Biofuels

, 8:166

First Online: 12 October 2015Received: 03 February 2015Accepted: 30 September 2015


BackgroundThe main challenge of cassava-based biobutanol production is to enhance the simultaneous saccharification and fermentation with high hyperamylolytic activity and butanol yield. Manipulation of cofactor e.g., Ca and NAD-PH levels as a potential tool to modulate carbon flux plays a key role in the cassava hydrolysis capacity and butanol productivity. Here, we aimed to develop a technology for enhancing butanol production with simultaneous hydrolysis of cassava a typical model as a non-cereal starchy material using a cofactor-dependent modulation method to maximize the production efficacy of biobutanol by Clostridium sp. stain BOH3.

ResultsSupplementing CaCO3 to the medium containing cassava significantly promotes activities of α-amylase responsible for cassava hydrolysis and butanol production due to the role of Ca cofactor-dependent pathway in conversion of cassava starch to reducing sugar and its buffering capacity. Also, after applying redox modulation with l-tryptophan a precursor as de novo synthesis of NADH and NADPH, the levels of cofactor NADH and NADPH increased significantly by 67 % in the native cofactor-dependent system of the wild-type Clostridium sp. stain BOH3. Increasing availability of NADH and NADPH improved activities of NADH- and NADPH-dependent butanol dehydrogenases, and thus could selectively open the valve of carbon flux toward the more reduced product, butanol, against the more oxidized acid or acetone products. By combining CaCO3 and l-tryptophan, 17.8 g-L butanol with a yield of 30 % and a productivity of 0.25 g-L h was obtained with a hydrolytic capacity of 88 % towards cassava in a defined medium. The metabolic patterns were shifted towards more reduced metabolites as reflected by higher butanol–acetone ratio 76 % and butanol–bioacid ratio 500 %.

ConclusionsThe strategy of altering enzyme cofactor supply may provide an alternative tool to enhance the stimulation of saccharification and fermentation in a cofactor-dependent production system. While genetic engineering focuses on strain improvement to enhance butanol production, cofactor technology can fully exploit the productivity of a strain and maximize the production efficiency.

KeywordsButanol Clostridium sp. Cofactor α-amylase Simultaneous saccharification and fermentation Cassava AbbreviationsABEacetone–butanol–ethanol

SSFsimultaneous saccharification and fermentation

NADHnicotinamide adenine dinucleotide hydrogen

NADPHnicotinamide adenine dinucleotide phosphate hydrogen

NADnicotinamide adenine dinucleotide

NADPnicotinamide adenine dinucleotide phosphate


BADHbutyraldehyde dehydrogenase

BDHAbutanol dehydrogenase I

BDHBbutanol dehydrogenase II

BDHCbutanol dehydrogenase III

AADalcohol-aldehyde dehydrogenase

AADCacetoacetate decarboxylase

RTreverse transcription

PCRpolymerase chain reaction

DCWdry cell weight

ODoptical density

Tinggang Li and Yu Yan contributed equally to this work

Electronic supplementary materialThe online version of this article doi:10.1186-s13068-015-0351-7 contains supplementary material, which is available to authorized users.

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Author: Tinggang Li - Yu Yan - Jianzhong He

Source: https://link.springer.com/

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