Molecular Characterization, Production and Optimization of Cellulase Producer and its Industrial Applications

Abstract

Background: Plant cell walls are composed primarily of cellulose, making it the most common organic polymer on Earth. The utilization of microorganisms to convert cellulose into valuable products has gained significant attention worldwide. This study aims to isolate novel cellulase-producing microorganisms and investigate the influence of dietary and physicochemical factors on cellulase production, as well as evaluate the potential applications of a selected isolated strain. Materials and Methods: The Bacillus amyloliquefaciens strain was identified as the most efficient producer of cellulase among various cellulase-producing bacterial strains isolated during the research. The morphological and biochemical characteristics of this strain were characterized. To determine the optimal conditions for cellulase production, various parameters such as raw substrates, carbon sources, nitrogen sources, temperature, inoculum age, inoculum size, agitation rate, incubation duration, and pH were investigated. Results: The study found that sugarcane bagasse, molasses, and paper pulp exhibited the highest cellulase production capacity, with respective enzyme activities of 0.97, 0.98, and 0.88 IU/mL. Maximum cellulase production occurred at a temperature of 37°C, an inoculum size of 2.5%, an inoculum age of 24 hours, an agitation rate of 120 rpm, an incubation time of 48 hours, and a pH of 7 with 1% peptone. The Bacillus amyloliquefaciens isolate was successfully applied to convert lignocellulosic waste into ethanol and fermentable sugars through saccharification and fermentation of cellulose, demonstrating its significant effectiveness. Conclusion: Bacillus amyloliquefaciens strain isolated in this study proved to be a highly efficient producer of cellulase. The optimized production media led to improved enzyme production, and the isolate showed promising results in the industrial application of converting cellulosic waste into ethanol. The 16s rRNA sequencing confirmed the identity of Bacillus amyloliquefaciens , which was deposited under the accession number MN081796 in the NCBI database. In this study we discussed the possible low-cost, enzymatic pretreatment methods of lignocellulosic material in order to use it as an efficient raw material for biofuel production. These findings contribute to the understanding and potential utilization of cellulose producing microorganisms in various biotechnological applications.