Heterologous Expression and Purification of ? Galactosidase Protein Using Affinity Chromatography

Authors

  • M. Z. Alam Shah Jalal University of Science and Technology, Sylhet
  • L. Ragionieri University of Aveiro
  • M. A. S. Santos University of Aveiro
  • A. Iqbal Shah Jalal University of Science and Technology, Sylhet-3114, Bangladesh

DOI:

https://doi.org/10.3329/jsr.v5i3.13820

Keywords:

?-galactosidase, Heterologous expression, GST tag, Affinity chromatography.

Abstract

Enzymes and other protein purification using recombinant DNA technology have become popular due to scarcity of natural protein. Saccharomyces cerevisiae is a demanding host, since it facilitates protein expression by its relative simplicity, safe organisms, inexpensive and has many properties of eukaryotic expression system. As an alternative host we express E. coli lacZ gene with GST tag in Saccharomyces cerevisiae and successfully purified from soluble extracts. The concentration of soluble GST-? galactosidase protein was approximately 0.57 mg/ml of elution buffer yielded from 50 ml yeast cell culture. The ?-galactosidase protein from insoluble extract was low due to the increasing solubility of GST tag.

Keywords: ?-galactosidase; Heterologous expression; GST tag; Affinity chromatography.

© 2013 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved.

doi: http://dx.doi.org/10.3329/jsr.v5i3.13820 J. Sci. Res. 5 (3), 499-513 (2013)

 

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Author Biographies

M. Z. Alam, Shah Jalal University of Science and Technology, Sylhet

Department of Genetic Engineering and Biotechnology

Assistant Professor

L. Ragionieri, University of Aveiro

Department of Biology and CESAM

Post Doctoral researcher

M. A. S. Santos, University of Aveiro

Department of Biology and CESAM

RNA biology laboratory

Associate Professor

A. Iqbal, Shah Jalal University of Science and Technology, Sylhet-3114, Bangladesh

Department of Genetic Engineering and Biotechnology

Assistant Professor

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Published

2013-08-28

How to Cite

Alam, M. Z., Ragionieri, L., Santos, M. A. S., & Iqbal, A. (2013). Heterologous Expression and Purification of ? Galactosidase Protein Using Affinity Chromatography. Journal of Scientific Research, 5(3), 499–513. https://doi.org/10.3329/jsr.v5i3.13820

Issue

Vol. 5 No. 3 (2013)

Section

Section B: Chemical and Biological Sciences

License

© Journal of Scientific Research

Creative Commons License

Articles published in the "Journal of Scientific Research" are Open Access articles under a Creative Commons Attribution-ShareAlike 4.0 International license (CC BY-SA 4.0). This license permits use, distribution and reproduction in any medium, provided the original work is properly cited and initial publication in this journal. In addition to that, users must provide a link to the license, indicate if changes are made and distribute using the same license as original if the original content has been remixed, transformed or built upon.