Isolation and functional characterization of phytase extracted from Klebsiella pneumoniae for enhanced growth of plants: A study of gene expression and in silico prediction.

The present study focused on isolating Klebsiella pneumoniae from sediment samples collected from poultry feeding and evaluating their ability to produce phytase. A total of 20 bacterial isolates were characterized using biochemical, and VITEK-2, among which five demonstrated positive phytase activity. To determine optimal conditions for phytase expression, different concentrations of wheat bran extract were tested. The most effective production was observed in media containing wheat bran extract at a pH of 6.5, incubated at 37 °C for 62 h. The purified phytase exhibited a molecular mass of approximately 51.0 kDa and showed significant enzymatic activity. Gene expression was further analyzed using real-time PCR to validate the optimal conditions (phyA gen - NC_012731). Additionally, in Silico molecular was conducted to investigate the interaction of the phytase enzyme with two protein targets: 5OTB (derived from Aspergillus niger) and 2AEE. Computational modeling using AutoDock 4 and Discovery Studio revealed that the phytase exhibited a more stable and stronger binding affinity with 5OTB, primarily due to favorable hydrogen bonding and electrostatic interactions. Generally, the findings suggest that K. pneumoniae is a viable candidate for phytase production, and the integration of molecular and computational approaches provides valuable insight into enzyme optimization and protein-ligand interactions.