Haemolytic and antibiofilm properties of haemocyanin purified from the haemolymph of Indian white shrimp Fenneropenaeus indicus

Abstract

Haemocyanin (Hc) is an important non-specific immune macromolecule present in the haemolymph of both mollusks and crustaceans. In the present study, Hc was purified from the haemolymph of Indian white shrimp Fenneropenaeus indicus by gel filtration chromatography and it exhibits a single band with a molecular weight of 74 kDa on SDS-PAGE. The X-ray diffraction (XRD) and High performance liquid chromatography (HPLC) result of purified Hc express single peak at 31.5° be a sign of crystalline nature and appear as a single peak with a retention time of 5.6 min signify the homogeneity nature of the protein respectively. The purified Hc exhibited haemolytic activity against chicken erythrocytes. The haemolytic activity of purified Hc in optimum conditions observed to be pH 6.0, temperature 40 °C, in the presence of calcium. As well purified Hc exhibited the antibiofilm activity against both Gram positive and Gram negative bacteria. Moreover, the haemolysis can be inhibited to different degrees by osmoprotectants of diverse molecular masses, signifying that it follows a colloid-osmotic mechanism. This study conclude that purified Hc from F. indicus remarkably possess haemolytic and antibiofilm activity.