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A small-scale investigation process for the Clostridium acetobutylicum production of butanol using high-energy carbon heavy ion irradiation.

ABSTRACT: Applied heavy ion irradiation technology and butanol industrial practices as a whole have been used as a strategy for the development of an attractive alternative to petroleum-based fuels. Clostridium acetobutylicum (C. acetobutylicum) strains are well documented as fermentation strains for the production of biobutanol. However, it has been reported that solvent production inhibits the growth of these strains, and the accumulation of acetate also inhibits biomass synthesis, rendering the production of butanol from acetone-butanol-ethanol (ABE) fermentation processes economically challenging. In this manuscript, we propose the use of high-energy carbon heavy ion irradiation from the Heavy Ion Research Facility in Lanzhou (HIRFL) to obtain a culture with an increased butanol yield. Our findings suggest that the use of a high-energy 12C6+ heavy ion irradiation dose of 45 Gy with an energy of 135 AMeV and ion pulses/levels of 106-108 favours ABE solvent production in an irradiated strain compared with the non-irradiated strain. The strategy reported in this manuscript may contribute to the development of a cost-effective butanol fermentation process that is competitive with similar fermentation processes.

SUBMITTER: Jiang TT

PROVIDER: S-EPMC6999245 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature

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A small-scale investigation process for the Clostridium acetobutylicum production of butanol using high-energy carbon heavy ion irradiation.

Jiang Ting-Ting TT Zhou Xiang X Liang Yan Y

Engineering in life sciences 20180920 10

Applied heavy ion irradiation technology and butanol industrial practices as a whole have been used as a strategy for the development of an attractive alternative to petroleum-based fuels. Clostridium acetobutylicum (C. acetobutylicum) strains are well documented as fermentation strains for the production of biobutanol. However, it has been reported that solvent production inhibits the growth of these strains, and the accumulation of acetate also inhibits biomass synthesis, renderi ...[more]

PMID: 32624866

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Project description:Butanol is an important industrial solvent and advanced biofuel that can be produced by biphasic fermentation by Clostridium acetobutylicum. It has been known that acetate and butyrate first formed during the acidogenic phase are reassimilated to form acetone-butanol-ethanol (cold channel). Butanol can also be formed directly from acetyl-coenzyme A (CoA) through butyryl-CoA (hot channel). However, little is known about the relative contributions of the two butanol-forming pathways. Here we report that the direct butanol-forming pathway is a better channel to optimize for butanol production through metabolic flux and mass balance analyses. Butanol production through the hot channel was maximized by simultaneous disruption of the pta and buk genes, encoding phosphotransacetylase and butyrate kinase, while the adhE1(D485G) gene, encoding a mutated aldehyde/alcohol dehydrogenase, was overexpressed. The ratio of butanol produced through the hot channel to that produced through the cold channel increased from 2.0 in the wild type to 18.8 in the engineered BEKW(pPthlAAD(**)) strain. By reinforcing the direct butanol-forming flux in C. acetobutylicum, 18.9 g/liter of butanol was produced, with a yield of 0.71 mol butanol/mol glucose by batch fermentation, levels which are 160% and 245% higher than those obtained with the wild type. By fed-batch culture of this engineered strain with in situ recovery, 585.3 g of butanol was produced from 1,861.9 g of glucose, with the yield of 0.76 mol butanol/mol glucose and productivity of 1.32 g/liter/h. Studies of two butanol-forming routes and their effects on butanol production in C. acetobutylicum described here will serve as a basis for further metabolic engineering of clostridia aimed toward developing a superior butanol producer. IMPORTANCE Renewable biofuel is one of the answers to solving the energy crisis and climate change problems. Butanol produced naturally by clostridia has superior liquid fuel characteristics and thus has the potential to replace gasoline. Due to the lack of efficient genetic manipulation tools, however, strain improvement has been rather slow. Furthermore, complex metabolic characteristics of acidogenesis followed by solventogenesis in this strain have hampered development of engineered clostridia having highly efficient and selective butanol production capability. Here we report for the first time the results of systems metabolic engineering studies of two butanol-forming routes and their relative importances in butanol production. Based on these findings, a metabolically engineered Clostridium acetobutylicum strain capable of producing butanol to a high titer with high yield and selectivity could be developed by reinforcing the direct butanol-forming flux.

Dataset Information

A small-scale investigation process for the Clostridium acetobutylicum production of butanol using high-energy carbon heavy ion irradiation.

Publications

A small-scale investigation process for the <i>Clostridium acetobutylicum</i> production of butanol using high-energy carbon heavy ion irradiation.

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