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నైరూప్య

Therapeutic Effectiveness of L-Methioninase as Anticancer Agent: In silico versus In vivo analysis

Bhawana Kharayat, Priyanka Singh*

L-methioninase has significant application as chemotherapeutic anticancer drug in pharmaceutical sector. The catalytic activity of L-methioninase has depleted the exogenous supply of L-methionine by arresting growth of methionine dependent cancer cells. In this study, L-methioninase was purified from fermented broth of new bacterial isolate Pseudomonas stutzeri MTCC 101 with molecular weight (45 kDa), Michaelis constant (K_m=36.5 mM) and maximum activity (V_max=500 μmole/min/mg protein). This enzyme was resolved into 291 peptide sequences after MALDI-TOF/MS spectroscopy. Molecular docking and simulation studies as in silico approach had confirmed its higher binding affinity towards L-threonine and L-methionine substrates. The substrate specificity test had confirmed its maximum activity for specific amino acid L-methionine substrate. The high conformational stability of its three dimensional model with L-methionine substrate was validated by performing in vitro analysis against cancer cell lines. The experiments were validated by performing in vitro analysis of this enzyme against growth of cancer cell lines. This purified enzyme showed good inhibitory effect against Hepatocellular cancer (Hep-G2) and human lung carcinoma (A549) with IC₅₀ value as 56 μg/ml and 53 μg/ml respectively. This enzyme could be could be effectively used for formulation of therapeutic anticancer drug in future.