• Energy Research

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.

[EN] The potential of microbial communities for efficient anaerobic conversion of raw microalgae was evaluated in this work. A long-term operated thermophilic digester was fed with three different Organic Loading Rates (OLR) (0.2, 0.3 and 0.4¿g·L¿1·d¿1) reaching 32¿41% biodegradability values. The microbial community analysis revealed a remarkable presence of microorganisms that exhibit high hydrolytic capabilities such as Thermotogae (~44.5%), Firmicutes (~17.6%) and Dictyoglomi, Aminicenantes, Atribacteria and Planctomycetes (below ~5.5%) phyla. The suggested metabolic role of these phyla highlights the importance of protein hydrolysis and fermentation when only degrading microalgae. The ecological analysis of the reactor suggests the implication of the novel group EM3 in fermentation and beta-oxidation pathways during microalgae conversion into methane. Scenedesmus spp. substrate and free ammonia concentration strongly shaped thermophilic reactor microbial structure. Partial Least Square Discriminant Analysis (PLS-DA) remarked the resilient role of minor groups related to Thermogutta, Armatimonadetes and Ruminococcaceae against a potential inhibitor like free ammonia. Towards low-cost biogas production from microalgae, this study reveals valuable information about thermophilic microorganisms that can strongly disrupt microalgae and remain in high solids retention anaerobic digesters. This research work has been supported by the Spanish Ministry of Economy and Competitiveness (MINECO, Project CTM2011-28595-C02-02) jointly with the European Regional Development Fund (ERDF), which are gratefully acknowledged. The authors are thankful to Fernando Fdz-Polanco research team (University of Valladolid, Spain) for providing the thermophilic sludge from their pilot plant to inoculate the bioreactor and Llúcia Martínez and Giusseppe D'Aria from FISABIO sequencing service (Valencia, Spain) for their technical support during the Illumina sequencing design.

Biogas production from lignocellulosic biomass is generally considered to be challenging due to the recalcitrant nature of this biomass. In this study, the recalcitrance of birch was reduced by applying steam-explosion (SE) pretreatment (210 °C and 10 min). Moreover, bioaugmentation with the cellulolytic bacterium Caldicellulosiruptor bescii was applied to possibly enhance the methane production from steam-exploded birch in an anaerobic digestion (AD) process under thermophilic conditions (62 °C).Overall, the combined SE and bioaugmentation enhanced the methane yield up to 140% compared to untreated birch, while SE alone contributed to the major share of methane enhancement by 118%. The best methane improvement of 140% on day 50 was observed in bottles fed with pretreated birch and bioaugmentation with lower dosages of C. bescii (2 and 5% of inoculum volume). The maximum methane production rate also increased from 4-mL CH4/g VS (volatile solids)/day for untreated birch to 9-14-mL CH4/g VS/day for steam-exploded birch with applied bioaugmentation. Bioaugmentation was particularly effective for increasing the initial methane production rate of the pretreated birch yielding 21-44% more methane than the pretreated birch without applied bioaugmentation. The extent of solubilization of the organic matter was increased by more than twofold when combined SE pretreatment and bioaugmentation was used in comparison with the methane production from untreated birch. The beneficial effects of SE and bioaugmentation on methane yield indicated that biomass recalcitrance and hydrolysis step are the limiting factors for efficient AD of lignocellulosic biomass. Microbial community analysis by 16S rRNA amplicon sequencing showed that the microbial community composition was altered by the pretreatment and bioaugmentation processes. Notably, the enhanced methane production by pretreatment and bioaugmentation was well correlated with the increase in abundance of key bacterial and archaeal communities, particularly the hydrolytic bacterium Caldicoprobacter, several members of syntrophic acetate oxidizing bacteria and the hydrogenotrophic Methanothermobacter.Our findings demonstrate the potential of combined SE and bioaugmentation for enhancing methane production from lignocellulosic biomass.

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.

Anaerobic degradation of enzymatically pretreated Chlorella vulgaris was aimed in an upflow anaerobic sludge blanket reactor (UASB) to evaluate the organic loading rate (OLR) effect on biomass valorization. Low OLRs resulted in high methane yields (171 mL CH4/g CODin) at low hydraulic retention time (HRT of 6 days). Firmicutes (35-43%), Bacteroidetes (17-18%) and Euryarchaeota (11%) dominated at low OLRs, promoting methanogenic activity. On the contrary, the highest OLRs resulted in low methane yield (86 mL CH4/gCODin) with a concomitant short-chain fatty acids (SCFAs) accumulation of 37% SCFAs-COD/CODin. The highest OLR decreased UASB reactor biodiversity, hampering Euryarchaeota population development (2.5%) and boosting Firmicutes (55%) and Proteobacteria (14%). These results demonstrated the suitability of UASB reactor configuration to reach high bioprocess efficiency for both, biogas and SCFAs production, with lower energetic and area requirements than those normally needed in continuous stirred tank reactors.

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.

AbstractVolatile fatty acids (VFAs) are regarded as building blocks with a wide range of applications, including biofuel production. The traditional anaerobic digestion used for biogas production can be alternatively employed for VFAs production. The present study aimed at maximizing VFAs productions from Chlorella vulgaris through anaerobic digestion by assessing the effect of stepwise organic loading rates (OLR) increases (3, 6, 9, 12 and 15 g COD L−1 d−1). The biological system was proven to be robust as organic matter conversion efficiency into VFAs increased from 0.30 ± 0.02 COD-VFAs/CODin at 3 g COD L−1 d−1 to 0.37 ± 0.02 COD-VFAs/CODin at 12 g COD L−1d−1. Even though, the hydrolytic step was similar for all studied scenario sCOD/tCOD = 0.52–0.58), the highest OLR (15 g COD L−1 d−1) did not show any further increase in VFAs conversion (0.29 ± 0.01 COD-VFAs/CODin). This fact suggested acidogenesis inhibition at 15 g COD L−1d−1. Butyric (23–32%), acetic (19–26%) and propionic acids (11–17%) were the most abundant bioproducts. Population dynamics analysis revealed microbial specialization, with a high presence of Firmicutes followed by Bacteroidetes. In addition, this investigation showed the microbial adaptation of Euryarchaeota species at the highest OLR (15 g COD L−1d−1), evidencing one of the main challenges in VFAs production (out-competition of archaea community to avoid product consumption). Stepwise OLR increase can be regarded as a tool to promote VFAs productions. However, acidogenic inhibition was reported at the highest OLR instead of the traditional hydrolytic barriers. The operational conditions imposed together with the high VFAs and ammonium concentrations might have affected the system yields. The relative abundance of Firmicutes (74%) and Bacteroidetes (20%), as main phyla, together with the reduction of Euryarchaeota phylum (0.5%) were found the best combination to promote organic matter conversion into VFAs.