Purification, characterization and biological functions of metalloprotein isolated from haemolymph of mud crab Scylla serrata (Forskal, 1775)

• Published in: International journal of biological macromolecules 2020-12, Vol.164, p.3901-3908
• Main Authors: Durairaj, Karthick Rajan, Saravanan, Kandasamy, Mohan, Kannan, Ravichandran, Samuthirapandian
• Format: Article
• Language: English
• Subjects: Animals; Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - isolation & purification; Anti-Bacterial Agents - pharmacology; Antibacterial and anticancer activity; Antineoplastic Agents - chemistry; Antineoplastic Agents - isolation & purification; Antineoplastic Agents - pharmacology; Apoptosis - drug effects; Brachyura - chemistry; Cell Survival - drug effects; Chemical Phenomena; Chromatography, High Pressure Liquid; Haemocyanin; Haemolymph; Hemolymph - chemistry; Metalloproteins - isolation & purification; Microbial Sensitivity Tests; Reactive Oxygen Species; Scylla serrata; Spectroscopy, Fourier Transform Infrared; Spectrum Analysis; X-Ray Diffraction
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2020.08.228

In recent years, an enormous number of naturally occurring biological macromolecules has been reported worldwide due to its antibacterial and anticancerous potential. Among them, in this study, the copper containing respiratory protein namely haemocyanin (HC) was isolated from the haemolymph of mud crab Scylla serrata. The isolated metalloprotein HC was purified using Sepharose column by gel filtration chromatography. The purified HC was separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and molecular weight of the protein was identified as 95 kDa. Fourier transform infrared spectrophotometer (FT-IR) and nuclear magnetic resonance (1H NMR) spectral data revealed the presence of amino acid constituents. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis based mass ion search exposed that the purified protein was HC. HC exhibited an in vitro bacteriostatic effects against the bacterial pathogens and also elevated ROS levels in the treated samples. The half maximal (50%) inhibitory concentration (IC50) of HC was found to be 80 μg/mL against lung cancer cells (A549). Our study collectively addressed the potential antibacterial and anti-cancerous activity of HC. The results obtained from this study suggest that HC can be used for therapeutical application in the near future. •The haemocyanin was isolated and purified from haemolymph of mud crab Scylla serrata.•Isolated haemocyanin was characterized and confirmed using FTIR, SDS-PAGE, XRD, NMR and LC-MS/MS.•The purified haemocyanin showed effective antibacterial activity against bacterial pathogens.•The anticancer properties of haemocyanin was evidenced against human A549 lung cancer cell lines.