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Purification, Characterization, and Mitogenic Potential of a Mucin-Specific Mycelial Lectin from Aspergillus sparsus

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Abstract

Lectins are carbohydrate binding proteins or glycoproteins that bind reversibly to specific carbohydrates present on the apposing cells, which is responsible for their ability to agglutinate red blood cells, lymphocytes, fibroblasts, etc. Due to their carbohydrate specificity, lectins have been used for purification and characterization of glycoproteins like antibodies, cytokines, tumor-associated glycoproteins, hormones, inhibitors, growth factors, various enzymes, membrane proteins (receptors), or even toxins and viruses. In the present study, a mycelial lectin from Aspergillus sparsus was purified, characterized, and evaluated for its mitogenic potential. Lectin could be effectively purified 17.8-fold in a single-step using affinity chromatography on mucin-sepharose column. Lectin migrated as a single band in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with an apparent molecular mass of 100.2 kDa. Lectin showed pH optima of 6.5–8.0, and optimum temperature was determined to be 20–30 °C. Lectin was stable within a pH range of 5.5–10.0 and showed fairly good thermostability. Lectin activity was unaffected in the presence of EDTA, while activity reduced upon interaction with denaturants. MTT assay revealed strong mitogenic potential of A. sparsus lectin at a concentration up to 100 μg/ml.

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Acknowledgments

Authors are thankful to the Department of Biotechnology, Punjabi University, Patiala, for providing necessary facilities to execute the work.

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  1. Carbohydrate and Protein Biotechnology Laboratory, Department of Biotechnology, Punjabi University, Patiala, 147 002, Punjab, India

    Ram Sarup Singh, Ranjeeta Bhari & Rumeet Kaur

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  1. Ram Sarup Singh
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  2. Ranjeeta Bhari
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  3. Rumeet Kaur
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Correspondence to Ram Sarup Singh.

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Singh, R.S., Bhari, R. & Kaur, R. Purification, Characterization, and Mitogenic Potential of a Mucin-Specific Mycelial Lectin from Aspergillus sparsus . Appl Biochem Biotechnol 175, 1938–1947 (2015). https://doi.org/10.1007/s12010-014-1419-8

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  • DOI: https://doi.org/10.1007/s12010-014-1419-8

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