Cellulosimicrobium cellulans strain:PW Genome sequencing and assembly. Cellulosimicrobium cellulans strain:PW

The European Union generated around 6.29 million tons of agricultural leftover from various crops, accounting for almost 17 percent of global agriculture residue. Soybeans, potatoes, oats, rye, maize, barley, and wheat are the most important agricultural crops in Poland, producing a total of 261012.77 tons of agricultural waste in 2019. These agro-wastes or lignocellulosic biomass are composed of carbohydrate polymers such as cellulose, hemicellulose, and lignin. As a result, microbes' ability to absorb organic materials is dependent on their ability to create the enzymes required for the breakdown of the substrate components, namely cellulose, hemicellulose, and lignin. Another novel enzyme system i.e., Lytic Polysaccharide Monooxygenase i.e. LPMO, is recently discovered for releasing a spectrum of cleavage products from their polymeric substrates cellulose, hemicellulose, or chitin. Different water and sludge samples were collected from the wastewater treatment plant of the paper industry in Szczecin, Poland. Each sample was further processed and inoculated on minimal medium plates containing carboxymethyl cellulose (CMC), pectin, starch, and xylan as the sole carbon source to isolate cellulolytic, pectinolytic, amylolytic, and xylanolytic bacteria respectively. Among all selected isolates, Cellulosimicrobium cellulans strain PW7 was selected on the basis of the quantitative estimation of cellulase, amylase, and pectinase enzyme activity. The whole-genome sequencing of the selected isolate was further performed. Apart from lignocellulolytic genes, the LPMO gene was found in the genome. In order to confirm whether the LPMO genes are functional, all three copies of this gene present in the bacterial genome were cloned into the pET30a vector and then transformed into an overproducing strain E.coli BL21.