Isolation, Computational Characterization and Structure Prediction of Chitinase Gene-Bbchit of Beauveria bassiana

Abstract

Beauveria bassiana is an entomopathogenic fungus that is used as a biopesticide for the control of many insect pests in agriculturally important crops. B. bassiana produces many extracellular hydrolytic enzymes that degrade the insect cuticle. Among them, the fungal exochitinase mainly assists in the infection by degrading the chitin present in the insect cuticle. To characterize the exochitinase from B. bassiana, the chitinase gene (Bbchit) of 1050bp was amplified from the genomic DNA through PCR using gene-specific primers. This chitinase gene was cloned into the pJET1.2 cloning vector, confirmed through sequencing. The sequence (GI; OP114061) has been deposited in the NCBI database and used for bioinformatics analysis. The various Physico-chemical properties of chitinase protein such as pI, EC, AI, GRAVY and instability index were predicted using primary structure analysis. Using MEGA11 software, the translated chitinase protein- Bbchit showed that it closely resembles with the QJE37897.1 (chitinase Chit37). Primary and secondary structure analysis helps to characterize the functionality of chitinase protein (Bbchit). The 3D structure of B. bassiana was predicted using Robetta, an abinitio prediction method and confirmed through in-silico using the SAVESv6.0 (PROCHECK) server. The isolated exochitinase gene from B. bassiana has been characterized for use as a bio-control agent for the control of insect pests in crop plants.