Optimization and biochemical characterization of a thermotolerant processive cellulase, PtCel1, of Parageobacillus thermoglucosidasius NBCB1.

Vermicomposting involves enrichment of microorganisms that are able to resist higher temperatures and perform simultaneous degradation of lignocellulose, and therefore, such microbial communities are a potential source of cellulolytic enzymes. This study aimed to optimize the production of a processive cellulase by Parageobacillus thermoglucosidasius NBCB1 isolated from vermicompost, under submerged fermentation of rice straw and to characterize the purified enzyme for industrial suitability. Cellulase production in basal medium (7.27 IU/mg) was enhanced to 61 IU/mg by One Factor At a Time approach, which was further improved to 78.46 IU/mg by genetic algorithm based artificial neural networking. The cellulase PtCel1 purified from bacterial culture showed a molecular weight of ≈33 kD, had activity on both crystalline (305 IU/mg) and amorphous (184 IU/mg) cellulose as substrates. It had pH and temperature optima of 5.5°C and 60°C, respectively, and retained 100% activity upon preincubation at 60°C for 1 h indicating thermostability. PtCel1 was tolerant to sodium dodecyl sulfate, glucose and mannose; and the various metal chlorides, such as sodium, magnesium, calcium and zinc, acted as inducers giving 77.54%, 45.15%, 61.10%, and 169.14% augmentation of activity, respectively. Its efficiency on cellulosic substrates and robustness against aforementioned chemical and thermal environment makes it suitable for industrial applications.

Functional characterization of thermotolerant microbial consortium for lignocellulolytic enzymes with central role of Firmicutes in rice straw depolymerization.

Rice (Oryza sativa L.) straw, an agricultural waste of high yield, is a sustainable source of fermentable sugars for biofuel and other chemicals. However, it shows recalcitrance to microbial catalysed depolymerization. We herein describe development of thermotolerant microbial consortium (RSV) from vermicompost with ability to degrade rice straw and analysis of its metagenome for bacterial diversity, and lignocellulolytic carbohydrate active enzymes (CAZymes) and their phylogenetic affiliations. RSV secretome exhibited cellulases and hemicellulases with higher activity at 60 °C. It catalysed depolymerization of chemical pretreated rice straw as revealed by scanning electron microscopy and saccharification yield of 460 mg g-1 rice straw. Microbial diversity of RSV was distinct from other compost habitats, with predominance of members of phyla Firmicutes, Proteobacteria and Bacteroidetes; and Pseudoclostridium, Thermoanaerobacterium, Chelatococcus and Algoriphagus being most abundant genera. RSV harboured 1389 CAZyme encoding ORFs of glycoside hydrolase, carbohydrate esterase, glycosyl transferase, carbohydrate binding module and auxiliary activity functions. Microorganisms of Firmicutes showed central role in lignocellulose deconstruction with importance in hemicellulose degradation; whereas representatives of Proteobacteria and Bacteroidetes contributed to cellulose and lignin degradation, respectively. RSV consortium could be a resource for mining thermotolerant cellulolytic bacteria or enzymes and studying their synergism in deconstruction of chemically pretreated rice straw.