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Purification of a Lectin from M. rubra Leaves Using Immobilized Metal Ion Affinity Chromatography and Its Characterization

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Abstract

Lectins represent a heterogeneous group of proteins/glycoproteins with unique carbohydrate specificity, with wide range of biomedical applications. The multi-step purification protocols generally used for purification of lectin result in a significant reduction in the final yield and activity. In the present study, Morus rubra lectin (MRL) was purified to homogeneity from the leaves using a single-step immobilized metal ion affinity chromatography (IMAC) procedure. The approximate molecular weight of purified MRL resolved as a single band on SDS-PAGE was 52 kDa. Final percentage yield of purified lectin by IMAC was calculated as 74.7 %. Purified MRL was specific to three sugars, galactose, d-galactosamine and N-acetyl-d-galactosamine, and rendered haemagglutination (HA) activity towards different human blood group RBCs. MRL showed stability over a wide range of temperature (up to 80 °C) and pH (4–11). Chelation of the lectin with EDTA did not alter HA which indicates that metal ion is not required for activity. In the presence of Fe2+, Ca2+, Zn2+, Ni2+, Mn2+, Na+ and K+, HA activity was reduced to 50 %, whereas the presence of trivalent metal ions (Fe3+ and Al3+) and Cu2+ did not affect the activity. In the presence of Mg2+ and Hg2+, only 25 % of HA activity remained.

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Abbreviations

MRL:

Morus rubra lectin

IMAC:

Immobilized metal ion affinity chromatography

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Acknowledgments

The authors thank Department of Science and Technology (DST), Government of India, for the financial support. The help rendered by Dr. Satheeshkumar P.K (CBST) during the preparation of the manuscript is gratefully acknowledged.

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  1. Centre for Bio-Separation Technology (CBST), VIT University, Vellore, Tamil Nadu, 632 014, India

    Thavamani Sureshkumar & Sulochana Priya

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  1. Thavamani Sureshkumar
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  2. Sulochana Priya
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Correspondence to Sulochana Priya.

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Sureshkumar, T., Priya, S. Purification of a Lectin from M. rubra Leaves Using Immobilized Metal Ion Affinity Chromatography and Its Characterization. Appl Biochem Biotechnol 168, 2257–2267 (2012). https://doi.org/10.1007/s12010-012-9934-y

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  • DOI: https://doi.org/10.1007/s12010-012-9934-y

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