Navigating the Complex Terrain of Methane Synthesis: Multienzyme Control Points and Data-Driven Strategies.

Anaerobic digestion is a crucial process in wastewater treatment, renowned for its sustainable biogas production capabilities and the simultaneous reduction of environmental pollution. However, dysregulation of vital biological processes and pathways can lead to reduced efficiency and suboptimal biogas output, which can be seen through low counts per million of sequences related to three critical control points for methane synthesis. Namely, tetrahydromethanopterin S-methyltransferase (MTR), methyl-coenzyme reductase M (MCR), and CoB/CoM heterodisulfide oxidoreductase (HDR) are the last reactions that must occur. This study leveraged sequencing data from NCBI's Sequence Read Archive (SRA) database whose sample origins were of two model full-scale biodigesters. By assembling a genome BBBAS3_2 (86.4% complete), this study was able to align, assemble, and follow expression patterns related to KEGG pathways and sample conditions. This study detected and estimated expression patterns of conserved (in methanogenic archae) alleles for electron cycling by the heterodisulfide reductase complex, methylation and demethylation by methyltransferases, and oxidation of active coenzymes A and B by methyl-coenzyme reductase to surveil counts of sequences at critical control points. Utilizing a streamlined cloud bioinformatics approach with Omicsbox and Kbase, this study bridges the division between data-intensive sequencing and innovative solutions for methane optimization.