No abstract

Chain elongation reactor microbiomes produce valuable medium-chain carboxylates (MCC) from non-sterile residual substrates where lactate is a relevant intermediate. Gas supply has been shown to impact chain elongation performance. In the present study, the effect of nitrogen gas (N2) supply on lactate metabolism, conversion rates, biomass growth, and microbiome composition was evaluated in a lactate-fed upflow anaerobic reactor with continuous or intermittent N2 gas supply. Successful MCC production was achieved with continuous N2 gas supply at low superficial gas velocities (SGV) of 0.22 m∙h−1. Supplying N2 at high SGV (>2 m∙h−1) either continuously (2.2 m∙h−1) or intermittently (3.6 m∙h−1) disrupted chain elongation, resulting in production of short-chain carboxylates (SCC), i.e., acetate, propionate, and n-butyrate. Caproiciproducens-dominated chain-elongating microbiomes enriched at low SGV were washed out at high SGV where Clostridium tyrobutyricum-dominated microbiomes thrived, by displaying higher lactate consumption rates. Suspended growth seemed to be dominant regardless of SGV and gas supply regime applied with no measurable sludge bed formed. The highest MCC production from lactate of 10 g COD∙L−1∙d−1 with electron selectivities of 72 ± 5%was obtained without N2 gas supply at a hydraulic retention time (HRT) of 1 day. The addition of 5 g∙L−1 of propionate did not inhibit chain elongation, but rather boosted lactate conversion rates towards MCC with n-heptylate reaching 1.8 g COD∙L−1∙d−1. N2 gas supply can be used for mixing purposes and to steer lactate metabolism to MCC or SCC production.

No abstract

Medium-chain carboxylates (MCC) derived from biomass biorefining are attractive biochemicals to uncouple the production of a wide array of products from the use of non-renewable sources. Biological conversion of biomass-derived lactate during secondary fermentation can be steered to produce a variety of MCC through chain elongation. We explored the effects of zero-valent iron nanoparticles (nZVI) and lactate enantiomers on substrate consumption, product formation and microbiome composition in batch lactate-based chain elongation. In abiotic tests, nZVI supported chemical hydrolysis of lactate oligomers present in concentrated lactic acid. In fermentation experiments, nZVI created favorable conditions for either chain-elongating or propionate-producing microbiomes in a dose-dependent manner. Improved lactate conversion rates and n-caproate production were promoted at 0.5–2 g nZVI⋅L–1 while propionate formation became relevant at ≥ 3.5 g nZVI⋅L–1. Even-chain carboxylates (n-butyrate) were produced when using enantiopure and racemic lactate with lactate conversion rates increased in nZVI presence (1 g⋅L–1). Consumption of hydrogen and carbon dioxide was observed late in the incubations and correlated with acetate formation or substrate conversion to elongated products in the presence of nZVI. Lactate racemization was observed during chain elongation while isomerization to D-lactate was detected during propionate formation. Clostridium luticellarii, Caproiciproducens, and Ruminococcaceae related species were associated with n-valerate and n-caproate production while propionate was likely produced through the acrylate pathway by Clostridium novyi. The enrichment of different potential n-butyrate producers (Clostridium tyrobutyricum, Lachnospiraceae, Oscillibacter, Sedimentibacter) was affected by nZVI presence and concentrations. Possible theories and mechanisms underlying the effects of nZVI on substrate conversion and microbiome composition are discussed. An outlook is provided to integrate (bio)electrochemical systems to recycle (n)ZVI and provide an alternative reducing power agent as durable control method.

Background Biotechnological processes for efficient resource recovery from residual materials rely on complex conversions carried out by reactor microbiomes. Chain elongation microbiomes produce valuable medium-chain carboxylates (MCC) that can be used as biobased starting materials in the chemical, agriculture and food industry. In this study, sunflower oil is used as an application-compatible solvent to accumulate microbially produced MCC during extractive lactate-based chain elongation. The MCC-enriched solvent is harvested as a potential novel product for direct application without further MCC purification, e.g., direct use for animal nutrition. Sunflower oil biocompatibility, in situ extraction performance and effects on chain elongation were evaluated in batch and continuous experiments. Microbial community composition and dynamics of continuous experiments were analyzed based on 16S rRNA gene sequencing data. Potential applications of MCC-enriched solvents along with future research directions are discussed. Results Sunflower oil showed high MCC extraction specificity and similar biocompatibility to oleyl alcohol in batch extractive fermentation of lactate and food waste. Continuous chain elongation microbiomes produced the MCC n-caproate (nC6) and n-caprylate (nC8) from l-lactate and acetate at pH 5.0 standing high undissociated n-caproic acid concentrations (3 g L−1). Extractive chain elongation with sunflower oil relieved apparent toxicity of MCC and production rates and selectivities reached maximum values of 5.16 ± 0.41 g nC6 L−1 d−1 (MCC: 11.5 g COD L−1 d−1) and 84 ± 5% (e− eq MCC per e− eq products), respectively. MCC were selectively enriched in sunflower oil to concentrations up to 72 g nC6 L−1 and 3 g nC8 L−1, equivalent to 8.3 wt% in MCC-enriched sunflower oil. Fermentation at pH 7.0 produced propionate and n-butyrate instead of MCC. Sunflower oil showed stable linoleic and oleic acids composition during extractive chain elongation regardless of pH conditions. Reactor microbiomes showed reduced diversity at pH 5.0 with MCC production linked to Caproiciproducens co-occurring with Clostridiumtyrobutyricum, Clostridiumluticellarii and Lactobacillus species. Abundant taxa at pH 7.0 were Anaerotignum, Lachnospiraceae and Sporoanaerobacter. Conclusions Sunflower oil is a suitable biobased solvent to selectively concentrate MCC. Extractive reactor microbiomes produced MCC with improved selectivity and production rate, while downstream processing complexity was reduced. Potential applications of MCC-enriched solvents may include feed, food and biofuels purposes.