Insilico Interaction study on Dispase Enzyme with Fibronectic Protein and Molecular Dynamic Study for functional stability

Geethanjali Murugesan; Dr.N.Hema Shenpagam

Volume : V, Issue : V, May - 2016

Insilico Interaction study on Dispase Enzyme with Fibronectic Protein and Molecular Dynamic Study for functional stability

Geethanjali Murugesan, Dr. N. Hema Shenpagam

Abstract :

Aspergillus niger (A.niger) is a filamentous fungi and an essential microorganism used in the field of biotechnology. A.niger is cultured for industrial production of many substances. This study aimed at producing neutral protease (Dispase) from fungal strain, isolated from 50-75 agricultural soil sources obtained in and around Coimbatore district. The fungal strains were isolated by means of Sabouraud Dextrose Agar(SDA), Potato Dextrose Agar(PDA) and Czapek Dox Agar methods. Lactophenol Cotton Blue (LCB) staining method was used to identify the fungal colony as Aspergillus sp. Species name was identified as Aspergillus niger by using internal transcribed spacer nucleotide sequence alignment method. Groundnut seed oil cake, cotton seed oil cake, rice straw, wheat an, rice an from different varieties of rice like ponni, jaya, jyothi, MO5 and ASD 17 and sugarcane bagasse were used as substrates for dispase production and their effects on incubation time, pH of the medium and temperature were observed. Dispase was purified through ammonium sulfate precipitation and ion-exchange chromatography. Its molecular weight was estimated to be 48 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE) and further characterization was carried out on Fourier transform infrared spectroscopy (FTIR) and Nuclear magnetic resonance (NMR) spectroscopy. Docking and simulation techniques were carried out in silico to find interaction between fionectin and dispase and to check the stability of the protein complex.