Summary
Syngas fermentation with acetogens is known to produce mainly acetate and ethanol efficiently. Co‐cultures with chain elongating bacteria making use of these products are a promising approach to produce longer‐chain alcohols. Synthetic co‐cultures with identical initial cell concentrations of
Clostridium carboxidivorans
and
Clostridium kluyveri
were studied in batch‐operated stirred‐tank bioreactors with continuous CO/CO
2
‐gassing and monitoring of the cell counts of both clostridia by flow cytometry after fluorescence
in situ
hybridization (FISH‐FC). At 800 mbar CO, chain elongation activity was observed at pH 6.0, although growth of
C. kluyveri
was restricted. Organic acids produced by
C. kluyveri
were reduced by
C. carboxidivorans
to the corresponding alcohols butanol and hexanol. This resulted in a threefold increase in final butanol concentration and enabled hexanol production compared with a mono‐culture of
C. carboxidivorans
. At 100 mbar CO, growth of
C. kluyveri
was improved; however, the capacity of
C. carboxidivorans
to form alcohols was reduced. Because of the accumulation of organic acids, a constant decay of
C. carboxidivorans
was observed. The measurement of individual cell concentrations in co‐culture established in this study may serve as an effective tool for knowledge‐based identification of optimum process conditions for enhanced formation of longer‐chain alcohols by clostridial co‐cultures.
The application of artificial microbial consortia for biotechnological production processes is an emerging field in research as it offers great potential for the improvement of established as well as the development of novel processes. In this review, we summarize recent highlights in the usage of various microbial consortia for the production of, for example, platform chemicals, biofuels, or pharmaceutical compounds. It aims to demonstrate the great potential of co-cultures by employing different organisms and interaction mechanisms and exploiting their respective advantages. Bacteria and yeasts often offer a broad spectrum of possible products, fungi enable the utilization of complex lignocellulosic substrates via enzyme secretion and hydrolysis, and microalgae can feature their abilities to fixate CO 2 through photosynthesis for other organisms as well as to form lipids as potential fuelstocks. However, the complexity of interactions between microbes require methods for observing population dynamics within the process and modern approaches such as modeling or automation for process development. After shortly discussing these interaction mechanisms, we aim to present a broad variety of successfully established co-culture processes to display the potential of artificial microbial consortia for the production of biotechnological products.
Syngas fermentation with clostridial co-cultures is promising for the conversion of CO to alcohols. A CO sensitivity study with Clostridium kluyveri monocultures in batch operated stirred-tank bioreactors revealed total growth inhibition of C. kluyveri already at 100 mbar CO, but stable biomass concentrations and ongoing chain elongation at 800 mbar CO. On/off-gassing with CO indicated a reversible inhibition of C. kluyveri. A continuous supply of sulfide led to increased autotrophic growth and ethanol formation by Clostridium carboxidivorans even at unfavorable low CO concentrations. Based on these results, a continuously operated cascade of two stirred-tank reactors was established with a synthetic co-culture of both Clostridia. An amount of 100 mbar CO and additional sulfide supply enabled growth and chain elongation in the first bioreactor, whereas 800 mbar CO resulted in an efficient reduction of organic acids and de-novo synthesis of C2-C6 alcohols in the second reactor. High alcohol/acid ratios of 4.5–9.1 (w/w) were achieved in the steady state of the cascade process, and the space-time yields of the alcohols produced were improved by factors of 1.9–5.3 compared to a batch process. Further improvement of continuous production of medium chain alcohols from CO may be possible by applying less CO-sensitive chain-elongating bacteria in co-cultures.
Acetobacterium woodii is known to produce mainly acetate from CO 2 and H 2 , but the production of higher value chemicals is desired for the bioeconomy.Using chain-elongating bacteria, synthetic co-cultures have the potential to produce longer-chained products such as caproic acid. In this study, we present first results for a successful autotrophic co-cultivation of A. woodii mutants and a Clostridium drakei wild-type strain in a stirred-tank bioreactor for the production of caproic acid from CO 2 and H 2 via the intermediate lactic acid. For autotrophic lactate production, a recombinant A. woodii strain with a deleted Lctdehydrogenase complex, which is encoded by the lctBCD genes, and an inserted D-lactate dehydrogenase (LdhD) originating from Leuconostoc mesenteroides, was used. Hydrogen for the process was supplied using an All-in-One electrode for in situ water electrolysis. Lactate concentrations as high as 0.5 g L -1 were achieved with the AiO-electrode, whereas 8.1 g L -1 lactate were produced with direct H 2 sparging in a stirred-tank bioreactor. Hydrogen limitation was identified in the AiO process. However, with cathode surface area enlargement or numbering-up of the electrode and on-demand hydrogen generation, this process has great potential for a true carbon-negative production of value chemicals from CO 2 .
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