Satish Babu Rajulapati

Optimization of L-asparaginase production from novel Enterobacter sp., by submerged fermentation using response surface methodology

ABSTRACT The culture conditions and nutritional rations influencing the production of extra cellular antileukemic enzyme by novel Enterobacter aerogenes KCTC2190/MTCC111 were optimized in shake-flask culture. Process variables like pH, temperature, incubation time, carbon and nitrogen sources, inducer concentration, and inoculum size were taken into account. In the present study, finest enzyme activity achieved by traditional one variable at a time method was 7.6 IU/mL which was a 2.6-fold increase compared to the initial value. Further, the L-asparaginase production was optimized using response surface methodology, and validated experimental result at optimized process variables gave 18.35 IU/mL of L-asparaginase activity, which is 2.4-times higher than the traditional optimization approach. The study explored the E. aerogenes MTCC111 as a potent and potential bacterial source for high yield of antileukemic drug.

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).

Optimization of Operating Conditions for the Production of Enhanced Antifungal Metabolites from Streptomonospora arabica VSM 25 by Full Factorial Design

Objectives: To execute the influence of the physico-chemical variables on the production of the bioactive metabolites by Streptomonospra arabica VSM-25 using Response Surface Methodology. Materials and Methods: An actinobacterium strain isolated from the deep sea marine sediment samples was identified as Streptomonospra arabica VSM-25 by conventional and molecular approaches. RSM was employed to study the impact of five variables, viz. incubation time, pH, temperature, galactose and peptone concentrations on the production of antifungal metabolites by VSM-25. Growth related production formation kinetics and substrate utilization in batch system was analysed using mathematical and unstructured kinetic models. Results: Statistical study showed that the incubation time, pH, Temperature, Concentration of galactose and peptone has a significant effect (p Conclusion: To the best of our knowledge this is the first report on the production of anti fungal metabolites from S. arabica using RSM and kinetic modelling studies which firmly support the application of RSM and kinetic modelling for optimization. The study may find potential application in rapid screening and production of novel drug molecules from unexploited natural sources. Key words: Streptomonospora arabica , Antimycotic Activity, Optimization, Kinetic Modelling, Response Surface Methodology, Bioactive Metabolites.