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Combination of Trace Metal to Improve Solventogenesis of Clostridium carboxidivorans P7 in Syngas Fermentation.

ABSTRACT: Higher alcohols such as butanol (C4 alcohol) and hexanol (C6 alcohol) are superior biofuels compared to ethanol. Clostridium carboxidivorans P7 is a typical acetogen capable of producing C4 and C6 alcohols natively. In this study, the composition of trace metals in culture medium was adjusted, and the effects of these adjustments on artificial syngas fermentation by C. carboxidivorans P7 were investigated. Nickel and ferrous ions were essential for growth and metabolite synthesis during syngas fermentation by P7. However, a decreased dose of molybdate improved alcohol fermentation performance by stimulating carbon fixation and solventogenesis. In response to the modified trace metal composition, cells grew to a maximum OD600 nm of 1.6 and accumulated ethanol and butanol to maximum concentrations of 2.0 and 1.0 g/L, respectively, in serum bottles. These yields were ten-fold higher than the yields generated using the original composition of trace metals. Furthermore, 0.5 g/L of hexanol was detected at the end of fermentation. The results from gene expression experiments examining genes related to carbon fixation and organic acid and solvent synthesis pathways revealed a dramatic up-regulation of the Wood-Ljungdahl pathway (WLP) gene cluster, the bcs gene cluster, and a putative CoA transferase and butanol dehydrogenase, thereby indicating that both de novo synthesis and acid re-assimilation contributed to the significantly elevated accumulation of higher alcohols. The bdh35 gene was speculated to be the key target for butanol synthesis during solventogenesis.

SUBMITTER: Han YF

PROVIDER: S-EPMC7546793 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Combination of Trace Metal to Improve Solventogenesis of Clostridium carboxidivorans P7 in Syngas Fermentation.

Han Yi-Fan YF Xie Bin-Tao BT Wu Guang-Xun GX Guo Ya-Qiong YQ Li De-Mao DM Huang Zhi-Yong ZY

Frontiers in microbiology 20200925

Higher alcohols such as butanol (C4 alcohol) and hexanol (C6 alcohol) are superior biofuels compared to ethanol. Clostridium carboxidivorans P7 is a typical acetogen capable of producing C4 and C6 alcohols natively. In this study, the composition of trace metals in culture medium was adjusted, and the effects of these adjustments on artificial syngas fermentation by C. carboxidivorans P7 were investigated. Nickel and ferrous ions were essential for growth and metabolite synthesis d ...[more]

PMID: 33101253

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Project description:Syngas fermentation processes with acetogenic bacteria like Clostridium carboxidivorans have been proven to be a promising approach for the conversion of CO-rich waste gases into short- and medium-chain alcohols. The challenge of synthesis gas impurities, on the other hand, has always been a major concern for establishing an industrial-scale process, since some of the trace components in waste gases, such as NH3, H2S, and NO x , can have inhibiting or even toxic effects on microbial growth and product formation. Thus, this study aims to identify the effects of the main trace impurities in syngas from gasification of biogenic residues by the supply of defined concentrations of trace impurities to the cultivation medium. Autotrophic gas fermentation studies were performed with C. carboxidivorans in batch-operated fully-controlled stirred-tank bioreactors with continuous gas supply (80% CO and 20% CO2). The syngas components NH3 and H2S had a positive effect on both growth and alcohol formation (ethanol, 1-butanol, and 1-hexanol). The maximum biomass concentration was increased by more than 50%, and the maximum ethanol concentration was more than doubled with 5.0 g L-1 NH4Cl or 1.0 g L-1 H2S provided by the addition of 2.2 g L-1 thioacetamide. The addition of the nitrogen oxide species nitrate and nitrite, on the other hand, reduced biomass growth as well as alcohol concentrations. Already, the supply of 0.1 g L-1 NaNO3 resulted in reduced growth and 25% reduction of the maximum ethanol concentration. The production of the longer chain alcohols 1-butanol and 1-hexanol was reduced as well. All NaNO2 concentrations tested showed a strong toxic effect on the metabolism of C. carboxidivorans, and neither CO consumption nor product formation was observed after addition. As a consequence, NO x components in syngas from the gasification of biogenic residues should be reduced by the gasification process and/or selectively removed from the syngas after gasification.

Project description:BackgroundThe product of current syngas fermentation systems is an ethanol/acetic acid mixture and the goal is to maximize ethanol recovery. However, ethanol currently has a relatively low market value and its separation from the fermentation broth is energy intensive. We can circumvent these disadvantages of ethanol production by converting the dilute ethanol/acetic acid mixture into products with longer carbon backbones, which are of higher value and are more easily extracted than ethanol. Chain elongation, which is the bioprocess in which ethanol is used to elongate short-chain carboxylic acids to medium-chain carboxylic acids (MCCAs), has been studied with pure cultures and open cultures of microbial consortia (microbiomes) with several different substrates. While upgrading syngas fermentation effluent has been studied with open cultures, to our knowledge, no study exists that has performed this with pure cultures.ResultsHere, pure cultures of Clostridium kluyveri were used in continuous bioreactors to convert ethanol/acetic acid mixtures into MCCAs. Besides changing the operating conditions in regards to substrate loading rates and composition, the effect of in-line product extraction, pH, and the use of real syngas fermentation effluent on production rates were tested. Increasing the organic loading rates resulted in proportionally higher production rates of n-caproic acid, which were up to 40 mM day-1 (4.64 g L-1 day-1) at carbon conversion efficiencies of 90% or higher. The production rates were similar for bioreactors with and without in-line product extraction. Furthermore, a lower ethanol/acetic acid ratio (3:1 instead of 10:1) enabled faster and more efficient n-caproic acid production. In addition, n-caprylic acid production was observed for the first time with C. kluyveri (up to 2.19 ± 0.34 mM in batch). Finally, the use of real effluent from syngas fermentation, without added yeast extract, but with added defined growth factors, did maintain similar production rates. Throughout the operating period, we observed that the metabolism of C. kluyveri was inhibited at a mildly acidic pH value of 5.5 compared to a pH value of 7.0, while reactor microbiomes perform successfully at mildly acidic conditions.ConclusionsClostridium kluyveri can be used as a biocatalyst to upgrade syngas fermentation effluent into MCCAs at pH values above 5.5.

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Combination of Trace Metal to Improve Solventogenesis of Clostridium carboxidivorans P7 in Syngas Fermentation.

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Combination of Trace Metal to Improve Solventogenesis of <i>Clostridium carboxidivorans</i> P7 in Syngas Fermentation.

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