The hunt for novel anticancer drugs with minimal side effects continues. This study strengthens the claim by providing biochemical evidences of anticancer activities of<em> Bauhinia variegata</em> bark extracts on lung carcinoma cells (A549 and H460). Bark extracts of <em>Bauhinia variegata</em> were prepared by different solvents using Soxhlet apparatus and tested for their antioxidant potential by DPPH assay. The lung cancer cell lines were treated with <em>Bauhinia variegata</em> bark extracts and viability of cells was measured by MTT assay; metastatic ability was determined through Scratch assay and effect on DNA integrity was shown by gel electrophoresis. The Petroleum Ether Bark Extract (PEBE) inhibits proliferation (A549, IC50 = 1.5 mg/ml) at 48 h treatment. DNA damage was observed in A549 cells by agarose gel electrophoresis. The Chloroform Bark Extract (CBE) inhibited proliferation of H460 (IC50 = 1 mg/ml) with DNA damage after 24 h treatment. Soft agar assay indicated decreased ability to form colonies and scratch test showed impaired migration of A549 and H460 to PEBE and CBE treatment respectively. Apoptosis was detected using fluorescent dye staining in A549 and H460 cells. Caspase 3 activity was increased significantly in A549 and H460 cells. PEBE and CBE decrease the mitochondrial membrane potential gradient (??m) of A549 and H460 cells respectively. This study categorically proves the cytotoxic activity of <em>Bauhinia variegata</em> bark extracts on A549 and H460 cells.

Escherichia coli DH10B has 1.1 kb ybdK gene which is responsible for encoding YbdK enzyme that possess a Gamma glutamyl cysteine synthetase activity. ybdK gene was ligated downstream of a constitutive derepressed lac promoter of a low copy number plasmid vector pBBR1MCS-2, giving rise to a recombinant plasmid pPAT. Sinorhizobium fredii NGR234 transformed with pPAT showed an augmented production of glutathione which in turn increased the production of cadmium sulphide nanoparticles to some extent. Also, a heterologous expression of YbdK in Sinorhizobium fredii NGR234 improved the oxidation status of bacterial cells which is confirmed by fluorescence microscopy images and fluorometry. Genetically modified (GM) cells stained by DCFDA showed a significant decrease in fluorescence compared to wild type (WT) cells. Physical and chemical properties of the nanoparticles produced by the pPAT transformed Sinorhizobium fredii NGR234 differed significantly compared to wild type (WT) Sinorhizobium fredii NGR234. Comparative analysis of the nanoparticles by FTIR and SEM analysis revealed the functional groups attached to nanoparticles and average nanoparticle size respectively. Nanoparticles synthesized by genetically modified (GM) bacteria were about 3 times smaller in size compared to those produced by wild type (WT) rhizobium. FTIR analysis revealed an augmented presence of peptide with the nanoparticles produced by GM bacteria compared to those produced by the WT bacteria. XRD data revealed that biosynthesized CdS nanoparticles are face centered crystalline particles which was confirmed by comparing the peaks to standard JCPDS data (JCPDS card no. 10-454).

Bauhinia variegata (Linn.), Caesalpiniaceae, known as Kachnar in Ayurvedic texts has been shown to have promising anticancer potential. The presence of bioactive molecules in plants which can be commercialized has led to research in knowing their chemical properties. Previous studies from our laboratory have validated the claim of anticancer property of Bauhinia variegata by providing biochemical evidence for the effect of petroleum ether bark extract (PEBE) and chloroform bark extract (CBE) on A549 and H460 lung cancer cell lines respectively. This work aimed at the application of various analytical techniques (thermal analysis, Fourier infrared transform spectroscopy and GC-MS) in the characterization of dried extracts of Bauhinia variegata with medicinal properties. TG-DSC-FTIR-GC-MS was applied to monitor the thermal stability and chemical properties of Bauhinia variegata bark extracts. The TG analysis of PEBE and CBE showed mass loss within three steps in different temperature ranges. The DSC curves for the dried extracts of Bauhinia variegata showed that thermal processes occur between 50 – 100°C for PEBE and 50-70°C for CBE. The FTIR spectroscopic studies of petroleum ether and chloroform bark extracts of Bauhinia variegata revealed different characteristic peak values with various functional groups in the extracts. The GC-MS study of PEBE revealed 18 compounds and CBE revealed 8 compounds. Thus, we have been successful in narrowing down to very few probable anticancer compounds from Bauhinia variegata bark extracts and also through this study, it was possible to substantiate the possibility of applying these techniques for use in the herbal medicines production from medicinal plants.