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Purification and Characterization of a Novel Thermostable Mycelial Lectin from Aspergillus terricola

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Abstract

Lectin has been isolated from mycelia of Aspergillus terricola by single step purification on porcine stomach mucin-Sepharose 4B affinity column. Lectin could be effectively purified with 75% recovery and 4.47-fold increase in specific activity. Lectin migrated as a single band on SDS-PAGE with an apparent molecular mass of 32.5 kDa. Sugar inhibition assay revealed that the lectin did not strongly interact with most carbohydrates and their derivatives tested while strong binding affinity to d-glucose, d-sucrose, N-acetyl-d-galactosamine, asialofetuin, porcine stomach mucin, and bovine submaxillary mucin was indicated. Neuraminidase and protease treatment to erythrocytes enhanced lectin titre. Lectin activity was stable within the pH range of 7.0–10.5. A. terricola lectin displayed remarkable thermostability and remained unaffected upon incubation at 70 °C for 2.5 h. Lectin did not require metal ions for its activity. Incubation with denaturants (urea, thiourea, and guanidine–HCl) substantially reduced lectin activity. Carbohydrate analysis revealed that it is a glycoprotein with 9.76% total sugars.

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Acknowledgment

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

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Authors and Affiliations

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

    Ram Sarup Singh, Ranjeeta Bhari, Hemant Preet Kaur & Monika Vig

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  1. Ram Sarup Singh
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  2. Ranjeeta Bhari
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  3. Hemant Preet Kaur
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  4. Monika Vig
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Correspondence to Ram Sarup Singh.

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Singh, R.S., Bhari, R., Kaur, H.P. et al. Purification and Characterization of a Novel Thermostable Mycelial Lectin from Aspergillus terricola . Appl Biochem Biotechnol 162, 1339–1349 (2010). https://doi.org/10.1007/s12010-009-8906-3

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