Production, Characterization and Cytotoxic Evaluation of Pyocyanin Pigment Extracted from Pseudomonas aeruginosa Isolated from Industrial Soil Resources
Indumathi Mullaiselvan, Vijayarani Kanagaraj, D. Baskaran, Murugan Balaraman, S. Meignanalakshmi
Abstract
Thirty-five isolates belonging to the various genus of bacteria family were isolated from 25 different soil samples from various industrial areas from different districts of Tamil Nadu. One out of them were identified as P.aeruginosa (P1) and screened for pyocyanin production using submerged fermentation. The Pyocyanin production reached µg/ml on modified pseudomonas broth. The identification of strain was confirmed by 16s rRNA, the similarity with other strains available in the database was 98% (FDAARGOS_767). P1 was accessed at gene bank with accession number CP041008.1. Identification of the PhzM gene encoding PhzM enzyme was carried out using gene-specific primer followed by PCR. 190bp size gene was amplified using PCR. PhzM gene was confirmed and similarity with (FDAARGOS_767) strain was 100% in the database. Pyocyanin was extracted by standard chloroform extraction method, purified by HPLC (C17 column) and characterized by UV-Vis absorption spectroscopy, Particle size, zeta potential, FT-IR and LC-MS. The SEM analysis of pyocyanin pigment showed the particle size of microencapsulated pyocyanin. The cytotoxic effect of pyocyanin was investigated using VERO cells and it showed a concentration-response relationship between the concentration and cell viability the cytotoxicity increased gradually with the increase of its concentration.