HPLC (SHIMADZU, SPD-10 A VP) with the silicon C18 column was used to separate and analyze PAHs under isocratic condition (solvent – acetonitrile:water (80:20) (v/v) detection wavelength – 254 nm). The flow rate of the mobile phase (acetonitrile) was maintained at 0.5 mL/min. The samples (20 μL) were injected to HPLC analyzer for the analysis of PAHs. Based on the retention time, the fractions were collected and further subjected to analysis. A Hewlett-Packard 689 gas chromatography equipped with

5973 mass spectrometer with HP-5MS (30 m × 0.25 mm I D × 0.25 μm) fused silica capillary column was used for the analysis. The column temperature program was set at 100 °C hold for selleck compound 1 min, 15 °C/min to 160 °C and 5 °C/min to 300 °C hold for 7 min. The GC injector was held isothermally at 280 °C with a splitless period of 3 min. Helium was used as the carrier gas, at a flow rate of 1 mL/min by using electronic pressure control. The GC–MS Anti-infection Compound Library mw interface temperature was maintained at 280 °C. The MS was operated in electron impact (EI) ionization mode with electron energy of 70 eV and the scan to determine appropriate masses for selected ion monitoring ranged from 50 to 500 amu (atom to mass unit). Standards from Sigma Aldrich were used for the PAH (anthracene) and their metabolites. GC–MS library search was

used to confirm the metabolites without standards. Genomic DNA (gDNA) of MTCC 5514 was extracted from using DNeasy Blood & Tissue Kit (Qiagen GmbH, Hilden, Germany) following the manufacturer’s protocol for Gram +ve bacteria. The 16S rRNA was PCR amplified using the universal primers 8F: 5′-AGAGTTTGATCCTGGCTCAG-3′ and 1492R: 5′-GGCTACCTTGTTACGACTT-3′ as described by Turner et al. [29]. Homology of the 16S rRNA sequence was compared with sequences available in databases using Blast from the National Center for Biotechnology Information [2] and the Ribosomal Database Project [7]. Alignment of obtained 16S rRNA sequence and sequences from the databases, were all trimmed

to the same length using CLUSTAL Omega algorithm [26]. The sequence details were already submitted to NCBI with the wide accession no. HM145910. The genes encoding the biosurfactant (licA3) and catechol 2,3 dioxygenase (C23O) of the chosen organism was studied Roflumilast and the details were summarized in the following paragraph. The primers for both, surfactant (licA3) and catechol 2,3 dioxygenase (C23O) genes were designed from earlier reports [6] and [27] and were synthesized at Eurofins Genomics India Pvt. Ltd. A portion of surfactant gene 0.26 kb (licA3) gene was pulled out from the genomic DNA using F: 5′- CAA AAG CGC ATC ATA CCA CGT TGA G – 3′ and R: 5′-AGC GGC ACA TAT TGA TGC GGT TC – 3′ primers, with 2.5 U of Taq DNA polymerase in a 25 μL reaction mixture, consisting of 100 ng of genomic DNA, 20 pmol of each primer, 200 μM dNTPs and 1X Taq buffer with 2 mM MgCl2.