Isolation, characterization and antimicrobial properties of hepatopancreas lectin of the freshwater crab Oziotelphusanaga

• Published in: Protein expression and purification 2024-10, Vol.222, p.106536, Article 106536
• Main Authors: Vargila, F., Bai, S. Mary Mettilda, Mary, J. Vinoliya Josephine, Citarasu, T.
• Format: Article
• Language: English
• Subjects: Antimicrobial activity; Fetuin; Hepatopancreas; Lectin; PRRs
• ISSN: 1046-5928; 1096-0279; 1096-0279
• DOI: 10.1016/j.pep.2024.106536

Lectins are versatile proteins that specifically recognize and interact with sugar moieties expressed on the cell surface. The potential of lectin in drug targeting and delivery has instigated interest to identify natural lectins. Crabs have been identified as a rich source of lectin because the innate immune system is activated on encounter of pathogens and helps in the production of lectin. Although the presence of lectins in crab's hemolymph is well documented, little information about lectin in hepatopancreas, a vital organ for immunity and digestion in crustaceans, is currently available. A calcium dependent lectin (75 kDa) was purified from the hepatopancreas of the freshwater crab Oziotelphusa naga by bioadsorption and fetuin linked Sepharose 4B affinity chromatography technique. The isolated hepatopancreas lectin is calcium dependent and maximum agglutination was observed with rabbit erythrocytes. The hemagglutinating activity of the hepatopancreas lectin was effectively inhibited by sugars, such as α-lactose, GlcNAc, trehalose and NeuAc. Compared to sialylated N-glycosylated proteins including transferrin and apo transferrin, sialylated O-glycosylated proteins like fetuin exhibited stronger inhibitory effect. The ability of erythrocytes to bind hepatopancreas lectin has been diminished by desialylation of the potent inhibitor, indicating the significance of sialic acid in lectin-ligand interactions. The purified hepatopancreas lectin showed a broad spectrum of antimicrobial activity against bacteria Staphylococcus aureus, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, E. coli and fungi Candida albicans and Aspergillus niger. The findings of this study demonstrate the significance of hepatopancreas lectin as a multifunctional defense protein that inhibits the growth of bacteria and fungi. •Purification of a hepatopancreas lectin from crab Oziotelphusa naga by bioadsorption and affinity chromatography.•The isolated hepatopancreas lectin is calcium dependent and exhibited maximum agglutination with rabbit erythrocytes.•Hemagglutination of hepatopancreas lectin was inhibited by sugar, α-lactose and glycoprotein, fetuin.•De-O-acetylation studies confirmed the O-acetyl specificity of the lectin.•The purified lectin demonstrated high antimicrobial activity against Staphylococcus aureus and Candida albicans.