Spider venom toxins, its purification, solubilization, and antimicrobial activity

Abstract

Aim: Purification of spider venom Crossopriza lyoni toxins on a Sepharose CL-6B 200 column and evaluation of antimicrobial susceptibility in infectious bacterial pathogenic strains. Materials and Methods: Solubilization of spider venom toxins in Triton X-100 (0.1%), phosphate-buffered saline, and trichloroacetic acid.For obtaining purified toxins/ peptides, and determination of their molecular weight of toxins, poison gland homogenate was loaded on a gel filtration column and 135 regular fractions were eluted at constant rate of 5mL/min. Antimicrobial potential was determined in serial microdilution assays and agar disc diffusion method in presence of spider toxin peptides. Results and Discussions: The elution pattern of purified and homogenized mouth poison glands displayed two major peaks at 280 nm. The first one was eluted in fraction No. 43–51 while the second one after fraction no. 61–90. From gel filtration chromatography, total yield of protein obtained was 67.3%. Low-molecular-weight peptides (6–64 kDa) were obtained in spider venom. These were subjected to employ for antimicrobial study that revealed possible interaction between different concentration ranges of spider venom toxins. In serial microdilution assays, low minimum inhibitory concentration values 3.75 μg/ml were obtained in the presence of spider toxins. By agar disc diffusion method, the diameter of inhibition zones in mm in the presence of spider toxin at a concentration range of 98.56–1.92 μg/ml in Escherichia coli, Salmonella typhi, and Vibrio cholera obtained was 17.86 ± 0.21 mm, 16.16 ± 0.21 mm, and 18.66 ± 0.21 mm, respectively. Conclusion: In the present investigation, spider toxins have generated lytic effect in bacterial cells that results in heavy cell death. No doubt spider toxins can be used as alternate of broad-spectrum antibiotics.