In silico modelling and molecular dynamics simulation studies on L-Asparaginase isolated from bacterial endophyte of Ocimum tenuiflorum

Bioactive compounds from endophytes have been used to treat various diseases. In the present study, L-Asparaginase producing endophytes were isolated from Ocimum tenuiflorum (Tulasi) from NIT Warangal, Telangana, India to treat Acute Lymphoblastic Leukemia (ALL) in which L-Asparagine (L-Asn) deamination plays a vital role in ALL treatment. 20 (bacteria and fungi) out of 35 endophytes have been screened for L-Asparaginase production using rapid plate assay technique, in which four strains produced high amounts of L-Asparaginase. 16 s Ribosomal RNA sequencing studies were performed, Bacillus stratosphericus organism was identified, and purified L-Asparaginase sequence has been tailored using MALDI/TOF (Applied Biosystems). The homology model was developed by using MODELLER 9.15v as the endophyte lacks crystal structure of L-Asparaginase enzyme and validated by dint of quality index tools. Docking studies were performed using iGemdock 2.1v. In comparison, free energy binding efficiency of receptor towards L-Asparagine (L-Asn) is good with lesser energy -71.6 kcal/mol in comparison to L-Glutamine (L-Gln) having -67.7 kcal/mol. In order to find the stability of the docked complexes in dynamics environment, molecular dynamics and simulation studies were performed using GROMACS V4.6.5. The trajectory analysis for 10 ns shows the better RMSD, RMSF, Rg and average number of hydrogen bonds for complex 1 (L-Asparaginase + L-Asn docked complex). Hence, complex 1 was found to be more stable than Complex 2 (L-Asparaginase + L-Gln docked complex).