Validation of biphenyl degradation pathway by polymerase chain reaction, peptide mass fingerprinting and enzyme analysis

Our previous studies showed, bacterium Aquamicrobium sp. SK-2 can degrade biphenyl and polychlorinated biphenyls (PCBs). In the present study, proteins involved in degradation was evaluated using various molecular biology methods. The gene bphC strain identified polymerase chain reaction method. Further key enzyme degradation, 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC) purified through anion exchange gel filtration chromatography, subsequently activity measured. N-terminal amino acid sequence of showed 92% homology with BphC Gram-negative bacteria (Pseudomonas KKS102, Comamonas testosterone, Burkholderiaceae bacterium, Delftia acidovorans, Achromobacter denitrificans). Fractions collected during protein purification were applied on SDS-PAGE gel. Significant bands selected gel, pieces cutout to analyze peptide mass fingerprinting (PMF) PMF method provided useful information about degradation. Apart from BphC, two other enzymes, benzoate dioxygenase catechol 2,3-dioxygenase which are process identified. results indicate that be degraded 2-hydroxymuconic-semialdehyde this result is accordance our study. Based all these we conclude a potential candidate for bioremediation contaminated places.