Biohydrogen production by lactate-driven dark fermentation of real organic wastes derived from solid waste treatment plants.

This study evaluated the hydrogen production potential through lactate-driven dark fermentation (LD-DF) of organic wastes from solid waste treatment plants, including the organic fraction of municipal solid waste (OFMSW), mixed sewage sludge, and two OFMSW leachates. In initial batch fermentations, only OFMSW supported a significant hydrogen yield (70.1 ± 7.7 NmL-H2/g-VS added) among the tested feedstocks. Lactate acted as an important hydrogen precursor, requiring the presence of carbohydrates for sequential two-step lactate-type fermentation. The impact of operational pH (5.5-6.5) and initial total solids (TS) concentration (5-12.5 % w/w) was also evaluated using OFMSW as substrate, obtaining hydrogen yields ranging from 6.6 to 55.9 NmL-H2/g-VSadded. The highest yield occurred at 6.5 pH and 7.5 % TS. The LD-DF pathway was indicated to be present under diverse pH and TS conditions, supported by employing a specialized microbial consortium capable of performing LD-DF, along with the observed changes in lactate levels during fermentation.

Assessment of the performance of a symbiotic microalgal-bacterial granular sludge reactor for the removal of nitrogen and organic carbon from dairy wastewater.

Cheese whey (CW) is a nutrient deficient dairy effluent, which requires external nutrient supplementation for aerobic treatment. CW, supplemented with ammonia, can be treated using aerobic granular sludge (AGS) in a sequencing batch reactor (SBR). AGS are aggregates of microbial origin that do not coagulate under reduced hydrodynamic shear and settle significantly faster than activated sludge flocs. However, granular instability, slow granulation start-up, high energy consumption and CO2 emission have been reported as the main limitations in bacterial AGS-SBR. Algal-bacterial granular systems have shown be an innovative alternative to improve these limitations. Unfortunately, algal-bacterial granular systems for the treatment of wastewaters with higher organic loads such as CW have been poorly studied. In this study, an algal-bacterial granular system implemented in a SBR (SBRAB) for the aerobic treatment of ammonia-supplemented CW wastewaters was investigated and compared with a bacterial granular reactor (SBRB). Mass balances were used to estimate carbon and nitrogen (N) assimilation, nitrification and denitrification in both set-ups. SBRB exhibited COD and ammonia removal of 100% and 94% respectively, high nitrification (89%) and simultaneous nitrification-denitrification (SND) of 23% leading to an inorganic N removal of 30%. The efficient algal-bacterial symbiosis in granular systems completely removed COD and ammonia (100%) present in the dairy wastewater. SBRAB microalgae growth could reduce about 20% of the CO2 emissions produced by bacterial oxidation of organic compounds according to estimates based on synthesis reactions of bacterial and algal biomass, in which the amount of assimilated N determined by mass balance was taken into account. A lower nitrification (75%) and minor loss of N by denitrifying activity (<5% Ng, SND 2%) was also encountered in SBRAB as a result of its higher biomass production, which could be used for the generation of value-added products such as biofertilizers and biostimulants.

Extraction pathways and purification strategies towards carminic acid as natural-based food colorant: A comprehensive review.

As a current trend of fabricating healthier products, food manufacturing companies seek for natural-based food colorant aiming to replace the synthetic ones, which apart from meeting sensorial and organoleptic aspects, they can also act as health promoters offering additional added value. Carminic acid is a natural based food colorant typically found in several insect taxa. However, there are current approaches which pursue the production of this natural pigment via biotechnological synthesis. To date, this colorant has been intensively applied in the manufacture of several food items. Unfortunately, one of the main limitations deals with the establishment of the right protocol of extraction and purification of this component since there is no report analyzing the main extraction techniques for obtaining carminic acid. Therefore, this review, for the first time, comprehensively analyzes the ongoing strategies and protocols proposed by scientists towards either extraction or purification of carminic acid from its origin source, and from biotechnological systems. Emphasis has been focused on the main findings dealing with extraction techniques and the relevant insights in the field. A detailed discussion is provided on the advantages and drawbacks of the reported extraction and purification methods, main solvents used and their key interactions with target molecules.

Continuous lactate-driven dark fermentation of restaurant food waste: Process characterization and new insights on transient feast/famine perturbations.

The effect of hydraulic retention time (HRT) on the continuous lactate-driven dark fermentation (LD-DF) of food waste (FW) was investigated. The robustness of the bioprocess against feast/famine perturbations was also explored. The stepwise HRT decrease from 24 to 16 and 12 h in a continuously stirred tank fermenter fed with simulated restaurant FW impacted on hydrogen production rate (HPR). The optimal HRT of 16 h supported a HPR of 4.2 L H2/L-d. Feast/famine perturbations caused by 12-h feeding interruptions led to a remarkable peak in HPR up to 19.2 L H2/L-d, albeit the process became stable at 4.3 L H2/L-d following perturbation. The occurrence of LD-DF throughout the operation was endorsed by metabolites analysis. Particularly, hydrogen production correlated positively with lactate consumption and butyrate production. Overall, the FW LD-DF process was highly sensitive but resilient against transient feast/famine perturbations, supporting high-rate HPRs under optimal HRTs.

Enhancement of biogas production rate from bioplastics by alkaline pretreatment.

The effect of alkali-based pretreatment on the methanization of bioplastics was investigated. The tested bioplastics included PHB [poly(3-hydroxybutyrate)], PHBH [poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)], PHBV [poly(3-hydroxybutyrate-co-3-hydroxyvalerate], PLA (polylactic acid), and a PLA/PCL [poly(caprolactone)] 80/20 blend. Prior to methanization tests, the powdered polymers (500-1000 µm) at a concentration of 50 g/L were subjected to alkaline pretreatment using NaOH 1 M for PLA and PLA/PCL, and NaOH 2 M for PHB-based materials. Following 7 days of pretreatment, the amount of solubilized carbon for PLA and its blend accounted for 92-98% of the total initial carbon, while lower carbon recoveries were recorded for most PHB-based materials (80-93%), as revealed by dissolved total organic carbon analysis. The pretreated bioplastics were then tested for biogas production by means of mesophilic biochemical methane potential tests. Compared to unpretreated PHBs, methanization rates of pretreated PHBs were accelerated by a factor of 2.7 to 9.1 with comparable (430 NmL CH4/g material feed) or slightly lower (15% in the case of PHBH) methane yields, despite featuring a 1.4-2.3 times longer lag phases. Both materials, PLA and the PLA/PCL blend, were only extensively digested when pretreated, yielding about 360-380 NmL CH4 per gram of material fed. Unpretreated PLA-based materials showed nearly zero methanization under the timeframe and experimental conditions tested. Overall, the results suggested that alkaline pretreatment can help to enhance the methanization kinetics of bioplastics.

Screening Enzymes That Can Depolymerize Commercial Biodegradable Polymers: Heterologous Expression of Fusarium solani Cutinase in Escherichia coli.

In recent years, a number of microbial enzymes capable of degrading plastics have been identified. Biocatalytic depolymerization mediated by enzymes has emerged as a potentially more efficient and environmentally friendly alternative to the currently employed methods for plastic treatment and recycling. However, the functional and systematic study of depolymerase enzymes with respect to the degradation of a series of plastic polymers in a single work has not been widely addressed at present. In this study, the ability of a set of enzymes (esterase, arylesterase and cutinase) to degrade commercial biodegradable polymers (PBS, PBAT, PHB, PHBH, PHBV, PCL, PLA and PLA/PCL) and the effect of pre-treatment methods on their degradation rate was assessed. The degradation products were identified and quantified by HPLC and LC-HRMS analysis. Out of the three enzymes, Fusarium solani cutinase (FsCut) showed the highest activity on grinded PBAT, PBS and PCL after 7 days of incubation. FsCut was engineered and heterologous expressed in Escherichia coli, which conferred the bacterium the capability of degrading solid discs of PBAT and to grow in PBS as the sole carbon source of the medium.

Unlocking the high-rate continuous performance of fermentative hydrogen bioproduction from fruit and vegetable residues by modulating hydraulic retention time.

Harnessing fruit-vegetable waste (FVW) as a resource to produce hydrogen via dark fermentation (DF) embraces the circular economy concept. However, there is still a need to upgrade continuous FVW-DF bioprocessing to enhance hydrogen production rates (HPR). This study aims to investigate the influence of the hydraulic retention time (HRT) on the DF of FVW by mixed culture. A stirred tank reactor under continuous mesophilic conditions was operated for 47 days with HRT stepwise reductions from 24 to 6 h, leading to organic loading rates between 47 and 188 g volatile solids (VS)/L-d. The optimum HRT of 9 h resulted in an unprecedented HPR from FVW of 11.8 NL/L-d, with a hydrogen yield of 95.6 NmL/g VS fed. Based on an overarching inspection of hydrogen production in conjunction with organic acids and carbohydrates analyses, it was hypothesized that the high FVW-to-biohydrogen conversion rate achieved was powered by lactate metabolism.

Influence of key operational parameters on biohydrogen production from fruit and vegetable waste via lactate-driven dark fermentation.

This study aims at investigating the influence of operational parameters on biohydrogen production from fruit-vegetable waste (FVW) via lactate-driven dark fermentation. Mesophilic batch fermentations were conducted at different pH (5.5, 6.0, 6.5, 7.0, and non-controlled), total solids (TS) contents (5, 7, and 9%) and initial cell biomass concentrations (18, 180, and 1800 mg VSS/L). Higher hydrogen yields and rates were attained with more neutral pH values and low TS concentrations, whereas higher biomass densities enabled higher production rates and avoided wide variations in hydrogen production. A marked lactate accumulation (still at neutral pH) in the fermentation broth was closely associated with hydrogen inhibition. In contrast, enhanced hydrogen productions matched with much lower lactate accumulations (even it was negligible in some fermentations) along with the acetate and butyrate co-production but not with carbohydrates removal. At pH 7, 5% TS, and 1800 mg VSS/L, 49.5 NmL-H2/g VSfed and 976.4 NmL-H2/L-h were attained.

Biogas bioconversion into poly(3-hydroxybutyrate) by a mixed microbial culture in a novel Taylor flow bioreactor.

Biogas-based biopolymer production represents an alternative biogas valorization route with potential to cut down plastic pollution and greenhouse gas emissions. This study investigated for the first time the continuous bioconversion of methane, contained in biogas, into poly(3-hydroxybutyrate) (PHB) by a mixed methanotrophic culture using an innovative high mass-transfer Taylor flow bioreactor. Following a hydrodynamic flow regime mapping, the influence of the gas residence time and the internal gas recirculation on CH4 abatement was assessed under non nutrient limiting conditions. Under optimal operational conditions (gas residence time of 60 min and internal gas recycling ratio of 17), the bioreactor was able to support a CH4 removal efficiency of 63.3%, a robust CH4 elimination capacity (17.2 g-CH4 m-3h-1) and a stable biomass concentration (1.0 g L-1). The simultaneous CH4 abatement and PHB synthesis was investigated under 24-h:24-h nitrogen feast/famine continuous operation. The cyclic nitrogen starvation and the Taylor flow imposed in the bioreactor resulted in a relatively constant biomass concentration of 0.6 g L-1 with PHB contents ranging from 11 to 32% w w-1 (on a dry weight basis), entailing an average PHB productivity of 5.9 g-PHB m-3 d-1 with an associated PHB yield of 19.8 mg-PHB g-CH4-1. Finally, the molecular analysis of the microbial population structure indicated that type II methanotrophs outcompeted non-PHB accumulating type I methanotrophs, with a heterotrophic-methanotrophic consortium enriched in Methylocystis, Hyphomicrobium, Rubinisphaeraceae SH PL14 and Pseudonocardia.

Production of volatile fatty acids (VFAs) from five commercial bioplastics via acidogenic fermentation.

The feasibility of producing volatile fatty acids (VFAs) from five commercial bioplastics via acidogenic fermentation by a non-pretreated anaerobic sludge was investigated. Mesophilic, anaerobic, acidogenic batch assays at 1, 10 and 20 g/L feed concentrations revealed the feasibility of producing VFAs from polyhydroxyalkanoates (PHA), i.e., PHB and PHBV, but not from PBS, PCL and PLA under the test conditions and time. However, only high PHA substrate concentrations (10-20 g/L) resulted in organic overloading and decreasing the pH of the culture broth down to 4-5, which in turn induced the accumulation of VFAs via kinetic imbalance between acidogenesis and methanogenesis. Gaseous carbon (C-CO2 and C-CH4) accounted for 8-35% of the total initial carbon, while C-VFAs represented 10-18%, mainly as acetate and butyrate. This study represents the first systematically assessed proof-of-concept to produce VFAs from PHA, which is key for the design of bioplastic-to-bioplastic recycling (bio)technologies.

Two-stage anaerobic digestion of food waste: Enhanced bioenergy production rate by steering lactate-type fermentation during hydrolysis-acidogenesis.

This study proposed a lactate-based two-stage anaerobic digestion (AD) process to enhance bioenergy production rate from food waste (FW) and investigated the effect of inoculum addition and culture pH on hydrolysis-acidogenesis and further methanization. A series of batch fermentations were performed with an enriched lactate-producing consortium and without inoculum addition under controlled (5.7) and uncontrolled pH (initial 6.7) conditions. The interplay between the studied factors dictated the fate of lactate, particularly if it is produced and accumulated in the fermentation broth or is consumed by butyrogenic bacteria. Only the self-fermentation of FW with uncontrolled pH resulted in lactate accumulation (0.2 g/g volatile solid (VS) fed) with limited off-gas production (0.32 NL/L) and VS losses (≈16%). Such lactate-rich broth was successfully digested through biochemical methane potential tests, resulting in a maximum bioenergy production rate of 2891 MJ/ton-VS fed per day, which was two-fold higher compared to that achieved by one-stage AD.

Biodegradation of bioplastics under aerobic and anaerobic aqueous conditions: Kinetics, carbon fate and particle size effect.

The biodegradation of PHB, PHBV, PBS, PBAT, PCL, PLA, and a PLA-PCL blend was compared under aerobic and anaerobic aqueous conditions assessing biodegradation kinetics, extent, carbon fate and particle size influence (in the range of 100-1000 µm). Under standard test conditions, PHB and PBHV were biodegraded anaerobically (83.9 ± 1.3% and 81.2 ± 1.7%, respectively) in 77 days or aerobically (83.0 ± 1.6% and 87.4 ± 7.5%) in 117 days, while PCL was only biodegraded (77.6 ± 2.4%) aerobically in 177 days. Apparent biomass growth accounted for 10 to 30.5% of the total initial carbon depending on the bioplastic and condition. Maximum aerobic and anaerobic biodegradation rates were improved up to 331 and 405%, respectively, at the lowest particle size tested (100-250 µm). This study highlights the usefulness of analysing biodegradation kinetics and carbon fate to improve both the development and testing of biodegradable materials, and waste treatments in the context of a circular bioeconomy.

Current evidence in high throughput ultrafiltration toward the purification of monoclonal antibodies (mAbs) and biotechnological protein-type molecules.

Pressure-driven membrane-based technologies, such as microfiltration (MF), ultrafiltration (UF), and nanofiltration (NF), have been successfully implemented in recovering different types of biomolecules and high-value-added compounds from various streams. Especially, UF membranes meet the requirements for separating specific bioproducts in downstream processes, e.g. monoclonal antibodies (mAbs), which are recognized as proteins produced mainly by plasma cells. According to the importance and functionality of the mAbs, their recovery is a current challenge with these bioseparations. Nevertheless, mAbs recovery using UF-assisted processes has been smartly performed over the last decade. To the best of our knowledge, there are no reviews of the reported developments using UF technology toward mAbs separation. Therefore, the goal of this paper is to collect and elucidate ongoing research studies implemented for the featured separation of mAbs and other biotechnological protein-type molecules (e.g. adenovirus serotype, extracellular vesicles, red fluorescent protein, cyanovirin-N, among others) via ultrafiltration-aided systems. The literature evidence (e.g. research papers, patents, etc.) has been analyzed and discussed according to the purpose of the study. Importantly, the relevant findings and novel approaches are discussed in detail. To finalize this document, the advantages, drawbacks, and guidelines in applying membrane-based techniques for such a recovery are presented.

Membrane-Based Harvesting Processes for Microalgae and Their Valuable-Related Molecules: A Review.

The interest in microalgae production deals with its role as the third generation of feedstock to recover renewable energy. Today, there is a need to analyze the ultimate research and advances in recovering the microalgae biomass from the culture medium. Therefore, this review brings the current research developments (over the last three years) in the field of harvesting microalgae using membrane-based technologies (including microfiltration, ultrafiltration and forward osmosis). Initially, the principles of membrane technologies are given to outline the main parameters influencing their operation. The main strategies adopted by the research community for the harvesting of microalgae using membranes are subsequently addressed, paying particular attention to the novel achievements made for improving filtration performance and alleviating fouling. Moreover, this contribution also gives an overview of the advantages of applying membrane technologies for the efficient extraction of the high added-value compounds in microalgae cells, such as lipids, proteins and carbohydrates, which together with the production of renewable biofuels could boost the development of more sustainable and cost-effective microalgae biorefineries.

Production of solid biofuels from organic waste in developing countries: A review from sustainability and economic feasibility perspectives.

The current increase in the world population and its energy demand promotes the study and implementation of cleaner energy forms since the traditional energy recovery systems are seriously affecting the environment. Biofuels and especially biomass or solid biofuels represent a sustainable energy source for developed and developing countries. This review aims to discuss the characteristics and advantages of solid biofuels, analyse the pretreatments and thermal treatments required to recover energy, and compare them with traditional fossil fuels. Other areas such as the sustainability and economic feasibility of solid biofuels are likewise addressed by explaining frequently used tools to evaluate the environmental impact as Life Cycle Assessment (LCA). Comparatively, more recent methodologies are examined as efforts for accomplishing sustainability in the biofuel industry, namely Life Cycle Sustainability Assessment (LCSA) and certification schemes like the Roundtable on Sustainable Biomaterials (RSB), the Inter-American Development Bank Sustainability Scorecard, and initiatives like the Roundtable for Sustainable Palm Oil (RSPO). Finally, it was revealed that the economic feasibility and competitiveness of solid biofuels differ among developing countries but represent a notable contribution to their energy matrix.

Inspired by nature: Microbial production, degradation and valorization of biodegradable bioplastics for life-cycle-engineered products.

The global environmental pollution by micro- and macro-plastics reveals the consequences of an extensive use of recalcitrant plastic products together with inappropriate waste management practices that fail to sufficiently recycle the broad types of conventional plastic waste. Biobased and biodegradable plastics are experiencing an uprising as their properties offer alternative waste management solutions for a more circular material economy. However, although the production of such bioplastics has advanced on scale, the end-of-life (EOL) (bio)technologies to promote circularity are lacking behind. While composting and biogas plants are the only managed EOL options today, advanced biotechnological recycling technologies for biodegradable bioplastics are still in an embryonic stage. Thus, developing efficient biotechnologies capable of transforming bioplastic waste into high-value chemical building blocks or into the constituents of the original polymer offers promising routes towards life-cycle-engineered products. This review aims at providing a comprehensive state-of-the-art overview of microbial-based processes involved in the complete lifecycle of bioplastics. The current trends in the bioplastic market, the beginning and EOL scenarios of bioplastics, and a critical discussion on the key factors and mechanisms governing microbial degradation are systematically presented. Also, a critical evaluation of terminology and international standards to quantify polymer biodegradability is provided together with the latest biotechnological recycling strategies, including the use of different pre-treatments for (bio)plastic waste. Finally, the challenges and future perspectives for the development of life-cycle-engineered biobased and biodegradable plastic products are discussed.

A review on the factors influencing biohydrogen production from lactate: The key to unlocking enhanced dark fermentative processes.

Dark fermentation (DF) is one of the most promising biological methods to produce bio-hydrogen and other value added bio-products from carbohydrate-rich wastes and wastewater. However, process instability and low hydrogen production yields and rates have been highlighted as the major bottlenecks preventing further development. Numerous studies have associated such concerns with the inhibitory activity of lactate-producing bacteria (LAB) against hydrogen producers. However, an increasing number of studies have also shown lactate-based metabolic pathways as the prevailing platform for hydrogen production. This opens a vast potential to develop new strategies to deal with the "Achilles heel" of DF - LAB overgrowth - while untapping high-performance DF. This review discusses the key factors influencing the lactate-driven hydrogen production, paying particular attention to substrate composition, the operating conditions, as well as the microbiota involved in the process and its potential functionality and related biochemical routes. The current limitations and future perspectives in the field are also presented.

Ongoing progress on novel nanocomposite membranes for the separation of heavy metals from contaminated water.

Membranes, as the primary separation element of membrane-based processes, have greatly attracted the attention of researchers in several water treatment applications, including wastewater treatment, water purification, water disinfection, toxic and non-toxic chemical molecules, heavy metals, among others. Today, the removal of heavy metals from water has become challenging, in which chemical engineers are approaching new materials in membrane technologies. Therefore, the current review elucidates the progress of using different concepts of membranes and potential novel materials for such separations, identifying that polymeric membranes can exhibit a removal efficiency from 77 up to 99%; while novel nanocomposite membranes are able to offer complete removal of heavy metals (up to 100%), together with unprecedented permeation rates (from 80 up to 1, 300 L m-2 h-1). Thereby, the review also addresses the highlighted literature survey of using polymeric and nanocomposite membranes for heavy metal removal, highlighting the relevant insights and denoted metal uptake mechanisms. Moreover, it gives up-to-date information related to those novel nanocomposite materials and their contribution to heavy metals separation. Finally, the concluding remarks, future perspectives, and strategies for new researchers in the field are given according to the recent findings of this comprehensive review.

Three-stage process for tequila vinasse valorization through sequential lactate, biohydrogen and methane production.

This study evaluated a novel three-stage process devoted to the cascade production of lactate, biohydrogen and methane from tequila vinasse (TV), with emphasis on attaining a high and stable biohydrogen production rate (HPR) by utilizing lactate as biohydrogen precursor. In the first stage, tailored operating conditions applied to a sequencing batch reactor were effective in sustaining a lactate concentration of 12.4 g/L, corresponding to 89% of the total organic acids produced. In the second stage, the stimulation of lactate-centered dark fermentation which entails the decoupling of biohydrogen production from carbohydrates utilization was an effective approach enabling stable biohydrogen production, having HPR fluctuations less than 10% with a maximum HPR of 12.3 L/L-d and a biohydrogen yield of 3.1 L/LTV. Finally, 1.6 L CH4/L-d and 6.5 L CH4/LTV were obtained when feeding the biohydrogen fermentation effluent to a third methanogenic stage, yielding a global energy recovery of 267.5 kJ/LTV.

Changes in performance and bacterial communities in a continuous biohydrogen-producing reactor subjected to substrate- and pH-induced perturbations.

The performance and microbial communities of a continuous dark fermentation reactor exposed to perturbations induced by substrate change and acidic pH shock were investigated. A mesophilic well-mixed reactor separately fed with two types of tequila vinasses (TVs) and lactose was operated at a fixed pH of 5.5, except during short-term pH (3.8) stress, for ~61 days at decreasing hydraulic retention times (HRTs) from 24 to 4 h. During the first ~23 days of operation with TV, a decrease in HRT down to 4 h resulted in the highest reported biohydrogen productivity from TV of 12.4 NL/L-d. It was shown that abrupt change in TV type (even with temporal feeding of lactose) and transient over-acidification impaired the normal operation of the reactor. However, it rapidly recovered from such disturbances, sustaining similar high-rate productivity to that previously encountered. Recovery was attributed to resistant and resilient microbial community features, as supported by molecular characterisation.