Editorial: The microbiology of the biogas process

[Excerpt] The world is facing unprecedent challenges, related with energy crisis and climate change. Intensification of renewable energy production, with special focus on sustainable biogas and biomethane, is one of the front-line topics today. Biogas/biomethane will play a role in the transition toward climate-neutral and secure energy system. Ensuring that biomethane is produced from organic waste/wastewater is essential to support circularity and sustainability. Scaling up biomethane production and assuring its economic competitiveness are current key challenges. Biogas generation occurs in natural and engineered environments, such as anaerobic bioreactors, and involves a cascade of reactions catalyzed by complex microbial communities. To unlock and boost the full potential of waste-based biomethane production, bioprocess optimization is needed, which requires deep knowledge on microbial diversity and physiology, as well as on the complex microbial interactions and metabolic networks occurring in biogas processes. This was the motivation for launching the Research Topic “The Microbiology of the Biogas Process,” which comprises six original research articles by 48 authors, addressing different facets of the theme and resorting to diverse approaches, reflecting the complexity of the topic. [...]This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit.info:eu-repo/semantics/publishedVersio

High pressure pre-treatments promote higher rate and degree of enzymatic hydrolysis of cellulose

The effect of high pressure (HP) pre-treatments on the subsequent enzymatic hydrolysis of cellulose from bleached kraft Eucalyptus globulus pulp by cellulase from Tricoderma viride was evaluated. Pressure pre-treatments of 300 and 400 MPa during 5–45 min, lead to both an increased rate and degree of hydrolysis, reaching values ranging from 1.5- to 1.9-fold, quantified by the formation of reducing sugars. Both the pressure and time under pressure influenced the enzymatic hydrosability of the cellulosic pulps, with the former being more important. The results indicate that the pressure pre-treatments promoted an increased accessibility of cellulose towards cellulase in the cell wall. The results obtained open promising possibilities, to contribute to overcome conventional limitations of enzymatic cellulose hydrolysis for the production of fermentable glucose, for the production of second generation bioethanol and chemicals by enhancement of both rate and yield of hydrolysis. The results are also of interest for the preparation of “pressure engineered” celullose with incremented tailored hydrolysis patterns

Insight into the role of facultative bacteria stimulated by micro-aeration in continuous bioreactors converting LCFA to methane

Conversion of unsaturated long chain fatty acids (LCFA) to methane in continuous bioreactors is not fully understood. Palmitate (C16:0) often accumulates during oleate (C18:1) biodegradation in methanogenic bioreactors, and the reason why this happens and which microorganisms catalyze this reaction remains unknown. Facultative anaerobic bacteria are frequently found in continuous reactors operated at high LCFA loads, but their function is unclear. To get more insight on the role of these bacteria, LCFA conversion was studied under microaerophilic conditions. For that, we compared bioreactors treating oleate-based wastewater (organic loading rates of 1 and 3 kg COD m-3 d-1), operated under different redox conditions (strictly anaerobic-AnR, -350 mV; microaerophilic-MaR, -250 mV). At the higher load, palmitate accumulated 7 times more in the MaR, where facultative anaerobes were more abundant, and only the biomass from this reactor could recover the methanogenic activity after a transient inhibition. In a second experiment, the abundance of facultative anaerobic bacteria, particularly Pseudomonas spp. (from which two strains were isolated), was strongly correlated (p<0.05) with palmitate-to-total LCFA percentage in the biofilm formed in a continuous plug flow reactor fed with very high loads of oleate. This work strongly suggests that micro-aeration stimulates the development of facultative bacteria that are critical for achieving LCFA conversion to methane in continuous bioreactors. Microbial networks and interactions of facultative and strict anaerobes in microbial communities should be considered in future studies.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER027462), the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte2020 − Programa Operacional Regional do Norte. Research of M.S.D. and A.J.C. were supported by the European Research Council under the European Union’s Seventh Framework Programme (FP/2007-2013)/ERC Grant Agreement No 323009.info:eu-repo/semantics/publishedVersio

OC33 Chromatographic approaches to study pine nut skin: exploitation of its composition and bioactivities

Pine nut skin (PNS) is an unexploited and uncharacterized by-product recovered during pine nut processing. The exploitation of by-products as sources of valuable compounds agrees with the current demand for the reduction of waste, and a transition to more sustainable production and consumption1. Therefore, PNS characterization and bioactive potentialities were assessed. The utilization of several chromatographic techniques allowed the characterization of PNS phenolic compounds (HPLC-DAD-UV and HPLC-DAD-ESI-MSn), and the carbohydrates quantification and structural characterization, after specific derivatization (GC-FID and GC-MS). PNS subcritical water extraction using microwave was optimized and the obtained extracts, separated into low-molecularweight (rich in phenolic compounds) and high-molecular-weight (rich in carbohydrates), were evaluated regarding their digestibility and prebiotic activity. The prebiotic potential was assessed by quantifying the short-chain fatty acids (HPLC-UV) produced after the in vitro faecal fermentation. HPLC-DAD-ESI-MSn allowed to identify PNS phenolic compounds, namely protocatechuic, p-coumaric, and caffeic acids, while HPLC-DAD-UV enabled the monomers identification of proanthocyanidins ((epi)catechins) and hydrolysable tannins (protocatechuic acid), after acid methanolysis. GC techniques allowed to disclose the polysaccharides structures (xyloglucans and pectic polysaccharides) and their degradation by microbiota. The fermentation of both extracts rich in phenolic compounds and rich in polysaccharides resulted in an increased production of acetic, propionic, and butyric acids when compared to the commercial prebiotic inulin, proposing these PNS extracts as prebiotic agents.The work was supported through the projects UIDB/50006/2020 and UIDP/50006/2020, funded by FCT/MCTES through national funds. Soraia P. Silva, Alondra González and Dalila Roupar thank FCT/MCTES and ESF through NORTE 2020 for their PhD grants (ref. SFRH/BD/136471/2018, SFRH/BD/06268/2021 and SFRH/DB/139884/2018 respectively). Elisabete Coelho thanks the research contract (CDL-CTTRI-88-ARH/2018 – REF. 049-88-ARH/2018) funded by national funds (OE), through FCT, in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19. Clarisse Nobre acknowledges FCT for the assistant research contract 2021.01234.CEECIND.info:eu-repo/semantics/publishedVersio

A wide perspective of carbon materials as catalysts for bioremediation of emerging pollutants and methanogenesis

Biotransformation of emerging pollutants under anoxic conditions can be accelerated by carbon materials (CM) acting as redox mediators. CM have been also extensively reported as facilitating external electron transfer in methanogenic processes. Here, different CM including magnetic carbon materials (C@MNP), were prepared, characterized and applied as RM on the biological reduction of Acid Orange 10 (AO10) and ciprofloxacin (CIP). CIP could be biologically removed in the presence of CNT and CNT@2%Fe, and AO10 decolourisation rates were 79-fold higher in the assays with CNT@2%Fe. The effect of carbon nanotubes (CNT) on the activity of several pure cultures of methanogens was also investigated, demonstrating that CNT could accelerate up to 17-fold the methane production rate. It is evident from this work that carbon materials with different chemical and textural characteristics can accelerate significantly bioremediation and methanogenic processes. The fact that concentrations as low as 0.1 g/L were used with positive effects, is remarkable in terms of economic feasibility of using CM as efficient catalysts in both processes.info:eu-repo/semantics/publishedVersio

Inhibition studies with 2-bromoethanesulfonate reveal a novel syntrophic relationship in anaerobic oleate degradation

Degradation of long-chain fatty acids (LCFAs) in methanogenic environments is a syntrophic process involving the activity of LCFA-degrading bacteria and hydrogen-utilizing methanogens. If methanogens are inhibited, other hydrogen scavengers are needed to achieve complete LCFA degradation. In this work, we developed two different oleate (C18:1 LCFA)-degrading anaerobic enrichment cultures, one methanogenic (ME) and another in which methanogenesis was inhibited (IE). Inhibition of methanogens was attained by adding a solution of 2-bromoethanesulfonate (BrES), which turned out to consist of a mixture of BrES and isethionate. Approximately 5 times faster oleate degradation was accomplished by the IE culture compared with the ME culture. A bacterium closely related to Syntrophomonas zehnderi (99\% 16S rRNA gene identity) was the main oleate degrader in both enrichments, in syntrophic relationship with hydrogenotrophic methanogens from the genera Methanobacterium and Methanoculleus (in ME culture) or with a bacterium closely related to Desulfovibrio aminophilus (in IE culture). A Desulfovibrio species was isolated, and its ability to utilize hydrogen was confirmed. This bacterium converted isethionate to acetate and sulfide, with or without hydrogen as electron donor. This bacterium also utilized BrES but only after 3 months of incubation. Our study shows that syntrophic oleate degradation can be coupled to desulfonation.IMPORTANCE In anaerobic treatment of complex wastewater containing fat, oils, and grease, high long-chain fatty acid (LCFA) concentrations may inhibit microbial communities, particularly those of methanogens. Here, we investigated if anaerobic degradation of LCFAs can proceed when methanogens are inhibited and in the absence of typical external electron acceptors, such as nitrate, iron, or sulfate. Inhibition studies were performed with the methanogenic inhibitor 2-bromoethanesulfonate (BrES). We noticed that, after autoclaving, BrES underwent partial hydrolysis and turned out to be a mixture of two sulfonates (BrES and isethionate). We found out that LCFA conversion proceeded faster in the assays where methanogenesis was inhibited, and that it was dependent on the utilization of isethionate. In this study, we report LCFA degradation coupled to desulfonation. Our results also showed that BrES can be utilized by anaerobic bacteria.Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of the UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145-FEDER-000004), funded by the European Regional Development Fund under the scope of Norte2020—Programa Operacional Regional do Norte. We also acknowledge Project MultiBiorefinery (SAICTPAC/0040/2015 [POCI-01-0145-FEDER-016403]), funded by Sistema de Apoio à Investigação Científica e Tecnológica (SAICT), Programas de Atividades Conjuntas (PAC), and the financial support of the European Research Council under the European Union Seventh Framework Programme (FP/2007-2013)/ERC (grant agreement 323009)info:eu-repo/semantics/publishedVersio

Therapeutic targeting of PD-1/PD-L1 blockade by novel small-molecule inhibitors recruits cytotoxic T cells into solid tumor microenvironment

© Author(s) (or their employer(s)) 2022.This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.Background: Inhibiting programmed cell death protein 1 (PD-1) or PD-ligand 1 (PD-L1) has shown exciting clinical outcomes in diverse human cancers. So far, only monoclonal antibodies are approved as PD-1/PD-L1 inhibitors. While significant clinical outcomes are observed on patients who respond to these therapeutics, a large proportion of the patients do not benefit from the currently available immune checkpoint inhibitors, which strongly emphasize the importance of developing new immunotherapeutic agents. Methods: In this study, we followed a transdisciplinary approach to discover novel small molecules that can modulate PD-1/PD-L1 interaction. To that end, we employed in silico analyses combined with in vitro, ex vivo, and in vivo experimental studies to assess the ability of novel compounds to modulate PD-1/PD-L1 interaction and enhance T-cell function. Results: Accordingly, in this study we report the identification of novel small molecules, which like anti-PD-L1/PD-1 antibodies, can stimulate human adaptive immune responses. Unlike these biological compounds, our newly-identified small molecules enabled an extensive infiltration of T lymphocytes into three-dimensional solid tumor models, and the recruitment of cytotoxic T lymphocytes to the tumor microenvironment in vivo, unveiling a unique potential to transform cancer immunotherapy. Conclusions: We identified a new promising family of small-molecule candidates that regulate the PD-L1/PD-1 signaling pathway, promoting an extensive infiltration of effector CD8 T cells to the tumor microenvironment.C and RCA are supported by the Fundação para a Ciência e a Tecnologia, Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES) (PhD grants PD/BD/128238/2016 (RCA) and SFRH/BD/131969/2017 (BC)). The authors thank the funding received from the European Structural & Investment Funds through the COMPETE Programme and from National Funds through FCT under the Programme grant LISBOA-01-0145-FEDER016405 - SAICTPAC/0019/2015 (HF and RCG). HFF and RCA received additional support from FCT-MCTES (UIDB/04138/2020, PTDC/BTM-SAL/4350/2021 and UTAPEXPL/NPN/0041/2021; EXPL/MED-QUI/1316/2021, respectively). The MultiNano@MBM project was supported by The Israeli Ministry of Health, and FCTMCTES, under the frame of EuroNanoMed-II (ENMed/0051/2016; HF and RS-F). HF and RS-F thank the generous financial support from ‘La Caixa’ Foundation under the framework of the Healthcare Research call 2019 (NanoPanther; LCF/PR/HR19/52160021), as well as CaixaImpulse (Co-Vax; LCF/TR/CD20/52700005). MP thanks the financial support from Liga Portuguesa Contra o Cancro – Nucleo Regional do Sul and ‘iNOVA4Health – UIDB/04462/2020’, a program financially supported by Fundação para a Ciência e Tecnologia/Ministério da Educação e Ciência. RS-F thanks the following funding agencies for their generous support: the European Research Council (ERC) Advanced Grant Agreement No. (835227)–3DBrainStrom, ERC PoC Grant Agreement no. 862580 – 3DCanPredict, The Israel Science Foundation (Grant No. 1969/18), The Melanoma Research Alliance (MRA Established Investigator Award n°615808), the Israel Cancer Research Fund (ICRF) Professorship award (n° PROF-18-682), and the Morris Kahn Foundation.info:eu-repo/semantics/publishedVersio

Conductive materials affect methanogenic activity of pure cultures of methanogens and syntrophic cocultures

info:eu-repo/semantics/publishedVersio

Acceleration of methane production by carbon nanotubes

Book of Abstracts of CEB Annual Meeting 2017[Excerpt] Carbon nanotubes and other conductive materials have been found to influence the rates of several anaerobic reactions. A range of different conductive carbon materials (CM) were reported to enhance methane production by anaerobic microbial communities. In most studies, the improvement of the overall process is attributed to the ability of these compounds to promote direct interspecies electron transfer (DIET) between bacteria, degrading more complex substrates, and methanogens, producing methane. The occurrence of DIET in the majority of these systems is not, however, proved and the effect of such conductive compounds on the activity of individual members, inside complex microbial communities, was never investigated. Thus, we herein present the results obtained when incubating pure cultures of methanogens, without any other microbial partner, in the presence of increasing concentrations of carbon nanotubes (CNT). Methane production from acetate, by the acetoclastic methanogens Methanosaeta concilii and Methanosarcina mazei, and from hydrogen plus carbon dioxide, by the hydrogenotrophic methanogens Methanospirillum hungatei and Methanobacterium formicicum, was accelerated, up to 17 times, in the presence of CNT [1]. Physical/chemical properties of the growth media changed in the presence of CNT, with redox potential decreasing with increasing CNT concentrations, and thus favouring methanogenesis. These findings show that CNT influences the microbial activity of methanogens in pure cultures and most likely this effect is extended to methanogens in complex communities as well, occurring in anaerobic bioreactors and in the environment. [...]info:eu-repo/semantics/publishedVersio