• Energy Research
• 2021-2025close

Short-chain fatty acids (SCFAs) are considered building blocks for bioproducts in the so-called carboxylate platform. These compounds can be sustainably produced via anaerobic fermentation (AF) of organic substrates, such as microalgae. However, SCFAs bioconversion efficiency is hampered by the hard cell wall of some microalgae. In this study, one thermal and two enzymatic pretreatments (carbohydrases and proteases) were employed to enhance Chlorella vulgaris biomass solubilization prior to AF. Pretreated and non-pretreated microalgae were assessed in continuous stirred tank reactors (CSTRs) for SCFAs production. Aiming to understand microorganisms' roles in AF depending on the employed substrate, not only bioconversion yields into SCFAs were evaluated but microbial communities were thoroughly characterized. Proteins were responsible for the inherent limitation of raw biomass conversion into SCFAs. Indeed, the proteolytic pretreatment resulted in the highest bioconversion (33.4% SCFAs-COD/CODin), displaying a 4-fold enhancement compared with raw biomass. Population dynamics revealed a microbial biodiversity loss along the AF regardless of the applied pretreatment, evidencing that the imposed operational conditions specialized the microbial community. In fact, a reduced abundance in Euryarchaeota phylum explained the low methanogenic activity, implying SCFAs accumulation. The bacterial community developed in the reactors fed with pretreated microalgae exhibited high acidogenic activities, being dominated by Firmicutes and Bacteroidetes. Firmicutes was by far the dominant phylum when using protease (65% relative abundance) while Bacteroidetes was prevailing in the reactor fed with carbohydrase-pretreated microalgae biomass (40% relative abundance). This fact indicated that the applied pretreatment and macromolecule solubilization have a strong effect on microbial distribution and therefore in SCFAs bioconversion yields.

AbstractIncreasing yeast robustness against lignocellulosic-derived inhibitors and insoluble solids in bioethanol production is essential for the transition to a bio-based economy. This work evaluates the effect exerted by insoluble solids on yeast tolerance to inhibitory compounds, which is crucial in high gravity processes. Adaptive laboratory evolution (ALE) was applied on a xylose-fermentingSaccharomyces cerevisiaestrain to simultaneously increase the tolerance to lignocellulosic inhibitors and insoluble solids. The evolved strain gave rise to a fivefold increase in bioethanol yield in fermentation experiments with high concentration of inhibitors and 10% (w/v) of water insoluble solids. This strain also produced 5% (P > 0.01) more ethanol than the parental in simultaneous saccharification and fermentation of steam-exploded wheat straw, mainly due to an increased xylose consumption. In response to the stress conditions (solids and inhibitors) imposed in ALE, cells induced the expression of genes related to cell wall integrity (SRL1,CWP2,WSC2andWSC4) and general stress response (e.g.,CDC5,DUN1,CTT1,GRE1), simultaneously repressing genes related to protein synthesis and iron transport and homeostasis (e.g.,FTR1,ARN1,FRE1), ultimately leading to the improved phenotype. These results contribute towards understanding molecular mechanisms that cells might use to convert lignocellulosic substrates effectively.

Abstract The organic fraction of municipal waste (OFMW), source-sorted (SS-OFMW) and non-sorted (NS-OFMW), was used as raw material for the sequential production of bioethanol and biogas. Non-isothermal and simultaneous saccharification and fermentation (NSSF) resulted in maximum ethanol concentrations of 51 g/L and 26 g/L for SS-OFMW and NS-OFMW samples, showing overall process yields of up to 80 % and 59 %, respectively, even without subjecting substrate to hydrothermal pretreatment. Subsequently, the solid residues resulting from the fermentation were further subjected to anaerobic digestion (AD), showing a methanogenic potential of 384 ± 6 mL CH4/g of volatile solids (VSin) and 322 ± 3 mL CH4/g VSin, respectively. These methane yields were similar or even higher to those obtained when using non-fermented OFMW substrates (SS-OFMW: 380 ± 18 mL CH4/g VSin and NS-OFMW: 239 ± 4 mL CH4/g VSin), highlighting NSSF as a beneficial step to enhance methane yields during AD. Overall, bioconversion of OFMW would benefit from coupling bioethanol and biogas production since the biogas produced might be further employed as bioenergy source to compensate operational costs.

Unspecific microorganisms consortia are normally used in anaerobic biodegradation of solid wastes. However, these consortia can be tuned to optimally obtain determined bioproducts. In this study, high value-added products and biogas were obtained via an innovative two-stage anaerobic bioprocess from microalgae biomass. The anaerobic fermentation (AF) entailed the production of short-chain fatty acids (SCFAs) and subsequently, only the solid spent of AF effluent was valorized for methane production via conventional anaerobic digestion (AD). Applied conditions in AF (25 °C, HRT 8 days) favored Firmicutes predominance (64%) enabling a conversion efficiency of 32.1% g SCFAs-COD/g CODin. Opposite, a wider microbial biodiversity was determined in the AD reactor (35 °C, HRT 20 days), being mainly composed by Firmicutes (28.6%), Euryarchaeota (17.7%) and Proteobacteria (15.3%). AD of the AF-solid spent reached 168.9 mL CH4 /g CODin. Strikingly, operational conditions imposed mediated a microbial specialization that maximized product output.

Lignocellulosic biomass utilization is challenging due to the presence of several carbon sources. Thus, microorganisms with different sugar preferences can be used in co-cultures to overcome this hurdle. This work addressed the simultaneous production of lactic acid (LA) from C5 sugars (i.e., xylose) and bioethanol from C6 (i. e., glucose/fructose) with Bacillus coagulans and Kluyveromyces marxianus, respectively. Sequential inoculation and co-inoculation of microorganisms were also compared. At pH 6, co-inoculation in synthetic media resulted in higher bioethanol (0.51 g/g) and LA (0.98 g/g) yields than sequential inoculation. Furthermore, when using lignocellulosic hydrolysates obtained after enzymatic hydrolysis of 20 % w/w pomegranate peels (PP), 92 % and 98 % of the theoretical maximum bioethanol and LA, respectively, were obtained. This study demonstrated the efficient bioethanol/LA co-generation despite the different optimum fermentation conditions of microorganisms and will pave the way to consider co-cultures for improving process efficiency in lignocellulosic biorefineries.

Anaerobic digestion (AD) of microalgae is an intriguing approach for bioenergy production. The scaling-up of AD presents a significant challenge due to the systematic efficiency losses related to process instabilities. To gain a comprehensive understanding of AD behavior, this study assessed a modified version of the anaerobic digestion model No1 (ADM1) + Contois kinetics to represent microalgae AD impacted by overloading. To this end, two new inhibition functions were implemented: inhibition by acetate for acidogenesis/acetogenesis and total volatile fatty acids for hydrolysis. This proposed ADM1 modification (including Contois kinetics) simulated AD behavior during the stable, disturbed and recovery periods, showing that the inhibition functions described in the original ADM1 cannot explain the AD performance under one of the most common perturbations at industrial scale (overloading). The findings underscore the importance of refining the inhibitions present in original ADM1 to better capture and predict the complexities of microalgae AD against overloading.

Dry anaerobic digestion (D-AD) generates nitrogen-rich effluents that are normally neglected in the circular bioeconomy. The high turbidity and ammonium content hamper nitrogen recovery from these effluents via biological processes, such as microalgae culture. The goal of this study was to demonstrate microalgae growth viability in high-strength D-AD effluents in order to recover nitrogen (N) as microalgae biomass. According to the experimental factorial design conducted in batch reactors, ammonium was identified as the critical inhibitory compound for microalgae growth while turbidity did not exhibit a significantly negative effect. Instead, turbidity resulted advantageous since it promoted high nitrogen uptake rates and biomass production. The presence of organic turbidity resulted in a positive effect that boosted Chlorella growth in a stream with higher ammonium (350 mg NH4+-N L-1) and turbidity (175 NTU) than the inhibition thresholds reported in the literature, reaching 98.7% of N recovery as microalgae biomass. When microalgae culture was scaled up in a photobioreactor operated in continuous mode, microalgae biomass was effectively produced while recovering 100% of N at a hydraulic retention time of 10 days. By imposing long exposure times and high turbidity, Chlorella adaptation to high-strength D-AD effluent resulted in high N uptake and biomass production. This study demonstrated not only the most influencing factor and the optimal NH4+-N and turbidity combination, but also the viability of using D-AD effluents as culture media for microalgae biomass production.