Parasporins, a class of non-insecticidal crystal proteins of Bacillus thuringiensis (Bt) are being explored as promising anticancer agents due to their specific toxicity to cancer cells. The present study has identified 25 Bt isolates harbouring parasporin genes from Western Ghats region, the hotspot of biodiversity in India. Among these, the isolate, KAU 41 (Kerala Agricultural University isolate 41) contained non-hemolytic homogenous crystals showing specific cytotoxicity towards cancer cells. SDS-PAGE analysis of this crystal, isolated by aqueous biphasic separation, revealed a 31 kDa sized peptide. The N-terminal sequence deciphered in BLAST analysis showed homology to a hypothetical Bt protein. Upon proteolysis, a 29 kDa active peptide was generated which exhibited heterogenic cytotoxic spectrum on various cancer cells. HeLa cells were highly susceptible to this peptide with IC 50 1 lg/mL and showed characteristics of apoptosis. RT-qPCR analysis revealed the overexpression of APAF1, caspase 3 and 9 by 14.9, 8 and 7.4 fold, respectively which indicates the activation of intrinsic pathway of apoptosis. However, at higher concentrations of peptide (greater than 3 lg/mL), necrotic death was prominent. The results suggest that the 31 kDa protein from Bt isolate, KAU 41 is a parasporin that may have high therapeutic potential.
Orchids are one of the main groups of ornamental plants commercially exploited. In the present study, we analyzed the diversity of bacterial community in Phalaenopsis root using metagenomic approach. The diversity of bacterial taxonomic category was assessed at different Operational Taxonomic Unit (OTU) levels using Ribosomal Database Project (RDP) pipeline and MG-RAST. At phylum level, Proteobacteria (61.34%) was the most dominant group followed by unclassified derived from bacteria (24.74%) and Actinobacteria (12.52%). Genus level analysis revealed the abundance of Rubrobacter, Pseudomonas and Acinetobacter. The study revealed that of the total species detected 50.83 per cent were unclassified, stressing the importance of metagenomics to assess the diversity of endophytes associated with orchid roots.
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