Comparative Study on Methods of Extraction of Protease from Marine Algae

ABSTRACT Protease is essential constituents of all forms of life on earth including fungi, plants, prokaryotes and animals. Proteases hydrolyze peptide bonds of proteins and they differ in their substrate specificity and ability to hydrolyze various peptide bonds. The present study was carried out to compare two methods of protease extraction namely, protein precipitation using ammonium sulphate and two phase extraction using PEG-4000 from seaweeds. The species studied were Enteromorpha intestinalis, Enteromorpha clathrata, Ulva lactuca, Gracilaria edulis. The protein content was estimated using Bradford method. The enzyme assay was estimated spectrophotometrically as well as by plate assay method. The results indicate that protease activity differs with species of seaweeds

ANTIOXIDANT AND ANTICANCER POTENTIALS OF BIOFABRICATED SILVER NANOPARTICLES

  Objective: The objective of this study is to explore a rapid, bio-inspired approach to synthesize silver nanoparticles (AgNPs) using aqueous Nardostachys jatamansi leaf extract and evaluate its antioxidant and cytotoxic activities on human colon carcinoma (HCT-116) cell lines.Methods: The biosynthesized nanoparticles were analyzed using ultraviolet-visible spectrophotometer, scanning electron microscope (SEM), energy dispersive X-ray, X-ray diffractometer (XRD), and Fourier transform infrared spectroscopy. Free radical scavenging and cytotoxic studies were carried out at different concentrations of AgNPs (20-100 μg/mL) using antioxidant 2,2-diphenyl-1-picrylhydrazil (DPPH) and mitochondrial function assay methods.Results: Surface plasmon resonance spectrum at 434 nm confirmed the formation of AgNPs. SEM images show biosynthesized AgNPs are mostly spherical shaped within the range of 30.0-58.7 nm. XRD analysis reveals the crystallographic face-centered cubic structure of the AgNPs. Thus, synthesized metal nanoparticles were tested for antioxidant activity by DPPH assay, and anticancer activity was validated by lactate dehydrogenase leakage assay. Significant antioxidant property was observed as compared to standard L-ascorbic acid. Further, AgNPs showed a linear dose-response relationship against HCT-116 cell lines with increasing concentration of AgNPs. At a concentration of 20 μg/mL, AgNPs were able to inhibit the cell line's growth by less than 9.8 ± 0.7%, whereas 100 μg/mL of AgNPs significantly inhibited the cell line's growth greater than 90.4 ± 0.25%.Conclusion: The synthesized AgNPs were found to be highly stable and had significant antioxidant and anticancer activity against HCT-116 cell lines. It has wide applications in the biomedical field and can be produced with eco-friendly, rapid scale-up, and easy downstream processing