Division of labor for substrate utilization in natural and synthetic microbial communities.

In natural ecosystems, microorganisms live in communities where each member interacts with the others and with the environment to efficiently utilise available resources. Division of Labor (DOL) is an evolutionary strategy that evolved by microbial communities to accomplish complex tasks. Current bio-based technologies could also benefit from DOL in microbial communities to construct robust microbial cell factories with expanded metabolic capabilities. Here, we review some remarkable examples of how DOL is used by natural microbial consortia to utilize a range of substrates. Also, we review the most recent studies towards engineering DOL to design synthetic consortia for efficient substrate utilization for bioproduction.

Integration of microbial electrochemical systems and photocatalysis for sustainable treatment of organic recalcitrant wastewaters: Main mechanisms, recent advances, and present prospects.

In recent years, microbial electrochemical systems (MESs) have demonstrated to be an environmentally friendly technology for wastewater treatment and simultaneous production of value-added products or energy. However, practical applications of MESs for the treatment of recalcitrant wastewater are limited by their low power output and slow rates of pollutant biodegradation. As a novel technology, hybrid MESs integrating biodegradation and photocatalysis have shown great potential to accelerate the degradation of bio-recalcitrant pollutants and increase the system output. In this review, we summarize recent advances of photo-assisted MESs for enhanced removal of recalcitrant pollutants, and present further discussion about the synergistic effect of biodegradation and photocatalysis. In addition, we analyse in detail different set-up configurations, discuss mechanisms of photo-enhanced extracellular electron transfer, and briefly present ongoing research cases. Finally, we highlight the current limitations and corresponding research gaps, and propose insights for future research.

Dark fermentation metabolic models to study strategies for hydrogen consumers inhibition.

A Flux Balance Analysis (FBA) metabolic model of dark fermentation was developed for anaerobic mixed cultures. In particular, the model was applied to evaluate the effect of a specific inoculum pre-treatment strategy, addition of waste frying oil (WFO) on H2-producing and H2-consuming metabolic pathways. Productions of volatile fatty acid (VFAs), CO2, H2 and CH4 measured through triplicate batch experiments, were used as constraints for the FBA model, to compute fluxes trough different metabolic pathways. FBA model could estimate the effect of pre-treatment with WFO on major microbial populations present in the mixed community (H2 producing bacteria, homoacetogen and methanogens). Results revealed that low concentrations of WFO did not completely inhibited hydrogenotrophic methanogens. FBA showed that acetoclastic methanogens were more sensitive to WFO, in comparison to hydrogenotrophic methanogens. The proposed model can be used to study H2 production by any other mixed microbial culture with similar substrates.

Pre-treating anaerobic mixed microflora with waste frying oil: A novel method to inhibit hydrogen consumption.

An innovative method was introduced to inhibit methanogenic H2 consumption during dark fermentative hydrogen production by anaerobic mixed cultures. Waste frying oil was used as an inhibitor for hydrogenotrophic methanogens. Simultaneous effect of waste frying oil concentrations (0-20 g/L) and initial pH (5.5, 6.5 and 7.5) on inhibition of methanogenic H2 consumption and enhancement of H2 accumulation were investigated using glucose as substrate. Enhanced hydrogen yields with decreased methane productions were observed with increasing the waste frying oil concentrations. On average, CH4 productions from glucose in the cultures received 10 g/L WFO were reduced by 88%. Increased WFO concentration up to 20 g/L led to negligible CH4 productions and in turn enhanced H2 yields. Hydrogen yields of 209.26, 195.35 and 185.60 mL/g glucoseadded were obtained for the cultures pre-treated with 20 g/L waste frying oil with initial pH of 5.5, 6.5 and 7.5 respectively. H2 production by pre-treated cultures was also studied using a synthetic food waste. Anaerobic mixed cultures were pre-treated with 10 g/L WFO and varying durations (0, 24 and 48 h). A H2 yield of 71.46 mL/g VS was obtained for cultures pre-treated with 10 g/L WFO for 48 h that was 475% higher than untreated control. This study suggests a novel and inexpensive approach for suppressing hydrogenotrophic methanogens during dark fermentative H2 production.

Pre-treatment technologies for dark fermentative hydrogen production: Current advances and future directions.

Hydrogen is regarded as a clean and non-carbon fuel and it has a higher energy content compared to carbon fuels. Dark fermentative hydrogen production from organic wastes is the most promising technology for commercialization among chemical and biological methods. Using mixed microflora is favored in terms of easier process control and substrate conversion efficiencies instead of pure cultures. However, mixed cultures should be first pre-treated in order to select sporulating hydrogen producing bacteria and suppress non-spore forming hydrogen consumers. Various inoculum pre-treatments have been used to enhance hydrogen production by dark fermentation including heat shock, acid or alkaline treatment, chemical inhibition, aeration, irradiation and inhibition by long chain fatty acids. Regarding substrate pre-treatment, that is performed with the aim of enhanced substrate biodegradability, thermal pre-treatment, pH adjustment using acid or base, microwave irradiation, sonication and biological treatment are the most commonly studied technologies. This article reviews the most investigated pre-treatment technologies applied for either inoculum or substrate prior to dark fermentation, the long-term effects of varying pre-treatment methods and the subsequently feasibility of each method for commercialization.

Effect of aerobic pre-treatment on hydrogen and methane production in a two-stage anaerobic digestion process using food waste with different compositions.

Aerobic pre-treatment was applied prior to two-stage anaerobic digestion process. Three different food wastes samples, namely carbohydrate rich, protein rich and lipid rich, were prepared as substrates. Effect of aerobic pre-treatment on hydrogen and methane production was studied. Pre-aeration of substrates showed no positive impact on hydrogen production in the first stage. All three categories of pre-aerated food wastes produced less hydrogen compared to samples without pre-aeration. In the second stage, methane production increased for aerated protein rich and carbohydrate rich samples. In addition, the lag phase for carbohydrate rich substrate was shorter for aerated samples. Aerated protein rich substrate yielded the best results among substrates for methane production, with a cumulative production of approximately 351ml/gVS. With regard to non-aerated substrates, lipid rich was the best substrate for CH4 production (263ml/gVS). Pre-aerated P substrate was the best in terms of total energy generation which amounted to 9.64kJ/gVS. This study revealed aerobic pre-treatment to be a promising option for use in achieving enhanced substrate conversion efficiencies and CH4 production in a two-stage AD process, particularly when the substrate contains high amounts of proteins.

Flux balance analysis of different carbon source fermentation with hydrogen producing Clostridium butyricum using Cell Net Analyzer.

A metabolic network model for Clostridium butyricum was developed using six different carbon sources (sucrose, fructose, galactose, mannose, trehalose and ribose) to study the fermentative H2 production. The model was used for investigation of H2 production and the ability of growth on different substrates to predict the maximum abilities for fermentative H2 production that each substrate can support. NADH fluxes were calculated by the model as an important cofactor affecting on H2 production. Butyrate and acetate production were used as model assumptions and biomass formation was chosen as the objective function for flux analysis calculations. Among the substrates selected, sucrose and trehalose supported the maximum growth and H2 yields. The Cell Net Analyzer metabolic network model developed for H2 estimation revealed good correlation with experimental data and could be further used to study the effect of environmental conditions and substrates concentration on H2 yield.