Scientific Reports of the National University of Life and Environmental Sciences of Ukraine

Suitability of crude glycerol as a substrate for biobutanol production

Olena Tigunova
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Abstract

Glycerol is a natural polyol formed as a major by-product during biodiesel production. The use of glycerol, which is uniquely utilised by Clostridium pasteurianum, for butanol production is highly promising but requires a thorough understanding and optimisation of the process. This study aimed to determine the composition of crude glycerol and evaluate its suitability as a substrate for butanol accumulation by Clostridium sp. UCM B-7570. During the research, a chromatographic method was used to determine the composition of crude glycerol and the solvent content in the culture liquid. An experimental approach was employed according to a developed scheme, incorporating microbiological methods (microorganism cultivation), biotechnological methods (strain cultivation under conditions resembling industrial settings, investigation of the solvent accumulation), and statistical methods for the mathematical processing of research results. A detailed study of the composition of different crude glycerol fractions showed that the initial glycerol layer contained a relatively low proportion of glycerol itself. The identified components accounted for more than half of the mass of the glycerol layer (51.6%). It was shown that the glycerol layer was found to contain only up to 20% glycerol and approximately 17% methanol, which is an inhibitor of microbial growth and development. It was determined that the highest butanol accumulation (9 g/L) occurred at a crude glycerol concentration of 35 g/L, while culture development was inhibited at 45 g/L. During the initial phase of fed‑batch cultivation of Clostridium sp. UCM B-7570, butanol accumulation remained unchanged. However, the subsequent fermentation of crude glycerol led to a twofold reduction in solvent by accumulation, ultimately resulting in complete inhibition of production by the eighth period, possibly due to the presence of methyl esters in the medium. To enhance butanol production technology, the use of sorbents such as activated carbon during fermentation is recommended. This study provides practical insights for biotechnologists and the demonstrated ability of Clostridium sp. UCM B-7570 to ferment crude glycerol for butanol production presents numerous research opportunities. These findings contribute to improving the feasibility of biobutanol production and advancing biomass-based industrial processes as viable alternatives to petroleum-derived products

Keywords

biofuel, waste, producer strains, fermentation, Clostridium

Suggested citation
Tigunova, O. (2024). Suitability of crude glycerol as a substrate for biobutanol production. Scientific Reports of the National University of Life and Environmental Sciences of Ukraine, 20(6),91-104. https://doi.org/10.31548/dopovidi/6.2024.91
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