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Lectin-Like Activity of Hemocyanin in Freshwater Prawn, Macrobrachium rosenbergii

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Abstract

Lectins are proteins that bind to the carbohydrate moieties on surface of bacteria, erythrocytes and other cells of invertebrates causing agglutination and mediate in recognition of foreign substances. In the present study, we isolated and characterized a lectin molecule present in the hemolymph of Macrobrachium rosenbergii, an important cultured freshwater prawn. Lectin in serum samples of adult prawns was assessed through hemagglutination (HA) test using rabbit RBC that showed a titre ranging from 16 to 64. This serum hemagglutinin was confirmed as a C-type lectin based on its dependency on calcium ions towards binding to rabbit RBCs. The hemagglutinin was also found to be stable at the pH range of 5.0–10.0 and temperature range of 10–40 °C. Of various sugars and glycoproteins tested in hemagglutination inhibition assay, the serum lectin was found specific only to N-acetylneuraminic acid and fetuin with respective minimum inhibitory concentrations at 50 mM and 0.31 mg/ml. Further, the lectin was purified by affinity chromatography on rabbit erythrocyte stroma, which showed hemagglutination with rabbit RBC. In electrophoretic analyses, the purified lectin showed one band with molecular weight of ~ 427 kDa in native gradient PAGE, and its two constituent polypeptide chains of ~ 81 and ~ 73 kDa in SDS-PAGE. These polypeptides were analysed in MALDI-TOF/TOF mass spectrometry and identified as hemocyanins. It was hence, concluded that hemocyanin in M. rosenbergii possesses lectin-like activity.

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Acknowledgements

The authors thank the Director, ICAR-Central Institute of Freshwater Aquaculture, Bhubaneswar for providing necessary facilities during the study. The financial support (Grant No. BT/PR12710/AAQ/3/713/2015) received from Department of Biotechnology, Govt. of India, New Delhi, India is also duly acknowledged.

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Authors and Affiliations

  1. ICAR–Central Institute of Freshwater Aquaculture, Bhubaneswar, 751002, India

    Jyotirmaya Mohanty, Sonalina Sahoo, Mohan R. Badhe, Bindu R. Pillai & Pramoda Kumar Sahoo

  2. Institute of Life Sciences, Bhubaneswar, 751023, India

    Amol R. Suryawanshi

  3. School of Biotech Sciences, Trident Academy of Creative Technology (TACT), Bhubaneswar, 751024, India

    Bharat Bhusan Patnaik

  4. P.G. Department of Bio-Science and Bio-Technology, Fakir Mohan University, Vyasa Vihar, Nuapadhi, Balasore, 756089, India

    Bharat Bhusan Patnaik

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  1. Jyotirmaya Mohanty
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  2. Sonalina Sahoo
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  3. Mohan R. Badhe
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  4. Bindu R. Pillai
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Correspondence to Jyotirmaya Mohanty.

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Mohanty, J., Sahoo, S., Badhe, M.R. et al. Lectin-Like Activity of Hemocyanin in Freshwater Prawn, Macrobrachium rosenbergii. Protein J 39, 358–365 (2020). https://doi.org/10.1007/s10930-020-09912-1

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