Anaerobic digestion (AD) is a microbially-driven process enabling energy production. Microorganisms are the core of anaerobic digesters and play an important role in the succession of hydrolysis, acidogenesis, acetogenesis, and methanogenesis processes. The diversity of participating microbial communities can provide new information on digester performance for biomass valorization and biofuel production. In this study anaerobic systems were used, operating under mesophilic conditions that realized biodegradation processes of waste wheat straw pretreated with NaOH—a renewable source for hydrogen and methane production. These processes could be managed and optimized for hydrogen and methane separately but combining them in a two-stage system can lead to higher yields and a positive energy balance. The aim of the study was to depict a process of biohydrogen production from lignocellulosic waste followed by a second one leading to the production of biomethane. Archaeal and bacterial consortia in a two-stage system operating with wheat straw were identified for the first time and the role of the most important representatives was elucidated. The mixed cultures were identified by the molecular-biological methods of metagenomics. The results showed that biohydrogen generation is most probably due to the presence of Proteiniphilum saccharofermentans, which was 28.2% to 45.4% of the microbial community in the first and the second bioreactor, respectively. Archaeal representatives belonging to Methanobacterium formicicum (0.71% of the community), Methanosarcina spelaei (0.03%), Methanothrix soehngenii (0.012%), and Methanobacterium beijingense (0.01%) were proven in the methane-generating reactor. The correlation between substrate degradation and biogas accumulation was calculated, together with the profile of fatty acids as intermediates produced during the processes. The hydrogen concentration in the biogas reached 14.43%, and the Methane concentration was 69%. Calculations of the energy yield during the two-stage process showed 1195.89 kWh·t−1 compared to a 361.62 kWh·t−1 cumulative yield of energy carrier for a one-stage process.

Anaerobic digestion (AD) is a widespread biological process treating organic waste for green energy production. In this study, wheat straw and corn stalks without any harsh preliminary treatment were collected as a renewable source to be employed in a laboratory-scale digester to produce biogas/biomethane. Processes parameters of temperature, pH, total solids, volatile solid, concentration of volatile fatty acids (VFA), and cellulose concentration, were followed. The volume of biogas produced was measured. The impact of organic loading was stated, showing that the process at 55 °C tolerated a higher substrate load, up to 45 g/L. Further substrate increase did not lead to biogas accumulation increase, probably due to inhibition or mass transfer limitations. After a 12-day anaerobic digestion process, cumulative volumes of biogas yields were 4.78 L for 1 L of the bioreactor working volume with substrate loading 30 g/L of wheat straw, 7.39 L for 40 g/L and 8.22 L for 45 g/L. The degree of biodegradation was calculated to be 68.9%, 74% and 72%, respectively. A fast, effective process for biogas production was developed from native wheat straw, with the highest quantity of daily biogas production occurring between day 2 and day 5. Biomethane concentration in the biogas was 60%. An analysis of bacterial diversity by metagenomics revealed that more than one third of bacteria belonged to class Clostridia (32.9%), followed by Bacteroidia (21.5%), Betaproteobacteria (11.2%), Gammaproteobacteria (6.1%), and Alphaproteobacteria (5%). The most prominent genera among them were Proteiniphilum, Proteiniborus, and Pseudomonas. Archaeal share was 1.37% of the microflora in the thermophilic bioreactor, as the genera Methanocorpusculum, Methanobacterium, Methanomassiliicoccus, Methanoculleus, and Methanosarcina were the most abundant. A knowledge of the microbiome residing in the anaerobic digester can be further used for the development of more effective processes in conjunction with theidentified consortium.

This study utilized a renewable energy source, agricultural waste, in anaerobic digestion (AD) at appropriate conditions to obtain biogas and biomethane as an energy carrier. Maize stalks underwent ultrasound (US) pre-treatment for better accessibility for microorganisms, as lignocelluloses have a stable structure, insoluble in water and resist both mechanical and enzymatic attack. The digestate after an anaerobic digestion process was used for cultivation of algae after adsorption with activated carbon for clarification. Photosynthetic microalgae have industrial and economic perspectives, so their low-cost cultivation has a great potential for many applications. The results showed the impact of US pre-treatment of maize stalks as a sole substrate and codigested with algal biomass. The total yields were 1116 cm 3 /L, 1350.5 cm 3 /L and 1293.25 cm 3 /L for the untreated, ultrasonically pre-treated and microwaved maize stalks. The possibility of accumulating algal biomass using anaerobic digestate as a medium was demonstrated. US pretreatment (400 W) showed high efficiency with respect to the extractives obtained per unit of energy input. Addition of 4 g/L of microalgal biomass as a co-substrate led to an increase in the biogas yield compared to native stalks. A small closed circle system, starting from anaerobic digestion of lignocellulosic substrates followed by microalgae cultivation in the digestate and subsequent return of microalgal biomass back in the bioreactor as a co-substrate was realized, encouraging circular economy. The suggested scheme is a simple and low-cost technology, as the substrate used is freely available and renewable, and algae proved to grow in a waste effluent as medium.