Anaerobic Two-Phase Co-Digestion for Renewable Energy Production: Estimating the Effect of Substrate Pretreatment, Hydraulic Retention Time and Participating Microbial Consortia

Abstract

Green and sustainable economies have recently become a key issue in long-term growth and well-being. Co-digestion of various waste materials in an eco-friendly way through biogas production has become the preferred method for their utilization and valorization. The possibility of hydrogen and methane yield maximization depends on the most suitable alkali reagent for pretreatment of waste lignocellulosic material, which was revealed in batch tests to determine the hydrogen production potential. The mixture for digestion consisted of pretreated wheat straw mixed with waste algal biomass in a ratio of 80:20 (w/w). The maximum hydrogen yield was achieved after applying sodium hydroxide thermoalkaline pretreatment, with a two-fold higher yield than the untreated control. Hydrogen production was stable and methane was not present in the resultant gas. The influence of the hydraulic retention time (HRT) on the maintenance of cascade installation was studied. The maximum daily concentration of hydrogen was achieved at an HRT of 2 days—42.5% H2—and the maximum concentration of methane was 56.1% at an HRT of 6 days. Accumulation of volatile fatty acids was registered in the first step and their depletion was noted in the second one. The obtained values of the cellulose content demonstrated that it was utilized by up to 2.75% in the methanogenic bioreactor at the end of the process. Metagenomics analyses revealed the bacteria Thermocaproicibacter melissae (44.9%) and Clostridium cellulosi (41.9%) participated in the consortium, accomplishing substrate hydrolysis and acidogenesis in the first stage. Less in abundance were Thermoanaerobacterium butyriciformans, Calorimonas adulescens, Pseudomonas aeruginosa and Anaerocolumna chitinilytica. Methanogenesis was performed by an archaeon closely related to Bathyarchaeota (99.5%) and Methanobacterium formicicum. The most abundant bacterial strains in the methanogenic fermenter were Abyssalbus ytuae (30%), Proteiniphilum acetatigenes (26%) and Ruficoccus amylovorans (13%).