Showing papers by "David P.B.T.B. Strik published in 2021"

Lactate Metabolism and Microbiome Composition Are Affected by Nitrogen Gas Supply in Continuous Lactate-Based Chain Elongation

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.

Cyclic Voltammetry is Invasive on Microbial Electrosynthesis

Abstract: Cyclic voltammetry (CV) is expected to cause changes in the biocathode composition, especially when using low scan rates. A recent finding stated that CV triggered further biocatalytic activity in microbial electrosynthesis systems (MES), leading to the aim of our study: to investigate the invasiveness of CV on MES. The present study confirms that a CO2 elongation MES biocathode composition changes during and right after the CV. Oxidation peaks differ over repeated CV-cycles while metal compounds and biomass were released in the biocatholyte. After CV, the current increased temporarily for up to 20 days and the metal compounds decreased from the biocatholyte solution. Further, the sole short application of open cell voltage was shown to shortly increase the current. Evidently CV affects the studied biocathode, which complicates the use of CV as an analysis technique in MESs. However, the positive effect CV has on biocathode current density may provide methods to boost reactor performance and maintain productivity.

Catalytic Cooperation between a Copper Oxide Electrocatalyst and a Microbial Community for Microbial Electrosynthesis.

Abstract: Electrocatalytic metals and microorganisms can be combined for CO2 conversion in microbial electrosynthesis (MES). However, a systematic investigation on the nature of interactions between metals and MES is still lacking. To investigate this nature, we integrated a copper electrocatalyst, converting CO2 to formate, with microorganisms, converting CO2 to acetate. A co-catalytic (i. e. metabolic) relationship was evident, as up to 140 mg L-1 of formate was produced solely by copper oxide, while formate was also evidently produced by copper and consumed by microorganisms producing acetate. Due to non-metabolic interactions, current density decreased by over 4 times, though acetate yield increased by 3.3 times. Despite the antimicrobial role of copper, biofilm formation was possible on a pure copper surface. Overall, we show for the first time that a CO2 -reducing copper electrocatalyst can be combined with MES under biological conditions, resulting in metabolic and non-metabolic interactions.

Open Culture Ethanol-Based Chain Elongation to Form Medium Chain Branched Carboxylates and Alcohols.

Abstract: Chain elongation fermentation allows for the synthesis of biobased chemicals from complex organic residue streams. To expand the product spectrum of chain elongation technology and its application range we investigated 1) how to increase selectivity towards branched chain elongation and 2) whether alternative branched carboxylates such as branched valerates can be used as electron acceptors. Elongation of isobutyrate elongation towards 4-methyl-pentanoate was achieved with a selectivity of 27% (of total products, based on carbon atoms) in a continuous system that operated under CO2 and acetate limited conditions. Increasing the CO2 load led to more in situ acetate formation that increased overall chain elongation rate but decreased the selectivity of branched chain elongation. A part of this acetate formation was related to direct ethanol oxidation that seemed to be thermodynamically coupled to hydrogenotrophic carboxylate reduction to corresponding alcohols. Several alcohols including isobutanol and n-hexanol were formed. The microbiome from the continuous reactor was also able to form small amounts of 5-methyl-hexanoate likely from 3-methyl-butanoate and ethanol as substrate in batch experiments. The highest achieved concentration of isoheptanoate was 6.4 ± 0.9 mM Carbon, or 118 ± 17 mg/L, which contributed for 7% to the total amount of products (based on carbon atoms). The formation of isoheptanoate was dependent on the isoform of branched valerate. With 3-methyl-butanoate as substrate 5-methylhexanoate was formed, whereas a racemic mixture of L/D 2-methyl-butanoate did not lead to an elongated product. When isobutyrate and isovalerate were added simultaneously as substrates there was a large preference for elongation of isobutyrate over isovalerate. Overall, this work showed that chain elongation microbiomes can be further adapted with supplement of branched-electron acceptors towards the formation of iso-caproate and iso-heptanoate as well as that longer chain alcohol formation can be stimulated.

nZVI Impacts Substrate Conversion and Microbiome Composition in Chain Elongation From D- and L-Lactate Substrates.

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.