• Energy Research

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.

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.

End-product accumulation during dark fermentation leads to process instability and hydrogen production inhibition. To overcome this constraint, microbial community adaptation to butyric acid can induce acid tolerance and thus enhance the hydrogen yields; however, adaptation and selection of appropriate microbial communities remains uncertain when dealing with complex substrates in a continuous fermentation mode. To address this question, a reactor fed in continuous mode with food waste (organic loading rate of 60 gVS·L·d−1; 12 h hydraulic retention time) was first stressed for 48 h with increasing concentrations of butyric acid (up to 8.7 g·L−1). Performances were compared with a control reactor (unstressed) for 13 days. During 6 days in a steady-state, the pre-stressed reactor produced 2.2 ± 0.2 LH2·L·d−1, which was 48% higher than in the control reactor (1.5 ± 0.2 LH2·L·d−1). The pretreatment also affected the metabolites’ distribution. The pre-stressed reactor presented a higher production of butyric acid (+44%) achieving up to 3.8 ± 0.3 g·L−1, a lower production of lactic acid (−56%), and an enhancement of substrate conversion (+9%). The performance improvement was attributed to the promotion of Clostridium guangxiense, a hydrogen -producer, with a relative abundance increasing from 22% in the unstressed reactor to 52% in the stressed reactor.

Biogas generation is the least complex technology to transform microalgae biomass into bioenergy. Since hydrolysis has been pointed out as the rate limiting stage of anaerobic digestion, the main challenge for an efficient biogas production is the optimization of cell wall disruption/hydrolysis. Among all tested pretreatments, enzymatic treatments were demonstrated not only very effective in disruption/hydrolysis but they also revealed the impact of microalgae macromolecular composition in the anaerobic process. Although carbohydrates have been traditionally recognized as the polymers responsible for the low microalgae digestibility, protease addition resulted in the highest organic matter solubilization and the highest methane production. However, protein solubilization during the pretreatment can result in anaerobic digestion inhibition due to the release of large amounts of ammonium nitrogen. The possible solutions to overcome these negative effects include the reduction of protein biomass levels by culturing the microalgae in low nitrogen media and the use of ammonia tolerant anaerobic inocula. Overall, this review is intended to evidence the relevance of microalgae proteins in different stages of anaerobic digestion, namely hydrolysis and methanogenesis.

Volatile fatty acids (VFAs) are chemical building blocks for industries, and are mainly produced via the petrochemical pathway. However, the anaerobic fermentation (AF) process gives a potential alternative to produce these organic acids using renewable resources. For this purpose, waste streams, such as microalgae biomass, might constitute a cost-effective feedstock to obtain VFAs. The present review is intended to summarize the inherent potential of microalgae biomass for VFA production. Different strategies, such as the use of pretreatments to the inoculum and the manipulation of operational conditions (pH, temperature, organic loading rate or hydraulic retention time) to promote VFA production from different microalgae strains, are discussed. Microbial structure analysis using microalgae biomass as a substrate is pointed out in order to further comprehend the roles of bacteria and archaea in the AF process. Finally, VFA applications in different industry fields are reviewed.