Isolation, Identification and Characterization of Cellulolytic Bacteria from Agricultural Waste Residues

Cellulases, the enzymes that catalyze the breakdown of cellulose, have emerged as critical players in the quest for sustainable energy solutions and the bioconversion of lignocellulosic biomass into valuable products. This study investigates the potential of novel, cellulase-producing bacteria isolated from soils amended with agricultural residues with the goal of improving biorefinery efficiency and reducing biofuel production costs. Through the selective screening of cellulose-degrading bacteria on carboxymethyl cellulose (CMC) agar, we identified a range of facultative anaerobic, Gram-negative, motile rod-shaped isolates. Morphological, cultural, and biochemical analyses suggested affiliations with the genera Enterobacter and Pseudomonas. We further elucidated the taxonomic identities of these isolates by amplifying and sequencing the 16S ribosomal ribonucleic acid (16S rRNA gene) using universal primers (27F and 1492R). The resulting 1465 bp nucleotide sequences were subjected to a comprehensive comparison with the NCBI nucleotide database, leading to the identification of Enterobacter cancerogenus, Pseudomonas aeruginosa, and Enterobacter cloacae. Phylogenetic analysis via the maximum likelihood method Phylogenetic analysis was conducted using the Maximum Likelihood method based on the Tamura-Nei model in MEGA 7.0 confirmed the evolutionary relationships among the isolates. Our findings demonstrate the cellulose-degrading potential of these strains based on qualitative screening assays. However, quantitative cellulase activity assays are required to fully evaluate their industrial applicability and enzyme production capacity.