Our previous studies showed, bacterium Aquamicrobium sp. SK-2 could degrade biphenyl and polychlorinatedbiphenyls (PCBs). In the present study, proteins involved in the biphenyl degradation was evaluatedusing various molecular biology methods. The gene bphC present in the strain SK-2 was identifiedusing the polymerase chain reaction method. Further the key enzyme in biphenyl degradation, 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC) was purified through anion exchange and gel filtration chromatography,subsequently the enzyme activity was measured. The N-terminal amino acid sequence ofthe purified enzyme showed 92% homology with BphC enzyme of Gram-negative bacteria(Pseudomonas sp. KKS102, Comamonas testosterone, Burkholderiaceae bacterium, Delftia acidovorans, andAchromobacter denitrificans). Fractions collected during protein purification were applied on SDS-PAGEgel. Significant bands were selected in SDS-PAGE gel, and the gel pieces were cut out to analyze the proteinsusing peptide mass fingerprinting (PMF) method. PMF method provided useful information aboutthe proteins involved in biphenyl degradation. Apart from BphC, two other enzymes, benzoate dioxygenaseand catechol 2,3-dioxygenase which were involved in biphenyl degradation process were identified.The results indicate that catechol can be degraded to 2-hydroxymuconic-semialdehyde and this result isin accordance with the results from our previous study. Based on all these results we can conclude thatthe strain SK-2 is a potential candidate for the bioremediation of biphenyl contaminated places._ 2021 The Authors. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.This is an open access article under the CC BY-NC-ND license
Validation of biphenyl degradation pathway by polymerase chain reaction, peptide mass fingerprinting and enzyme analysis
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