Preparation, Purification, and Secondary Structure Determination of Bacillus circulans Xylanase. A Modular Laboratory Incorporating Aspects of Molecular Biology, Biochemistry, and Biophysical Chemistry
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Sal Russo [1] ; Lisa Gentile [1]
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[1] Western Washington University
Western Washington University
Estados Unidos
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- Localización: Journal of chemical education, ISSN 0021-9584, Vol. 83, Nº 12, 2006, pág. 1850
- Idioma: inglés
- Texto completo no disponible (Saber más ...)
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Resumen
An integrated six-week module that allows students to develop biochemistry laboratory techniques in the context of a discovery-oriented project has been developed. This module, suitable for undergraduate third- or fourth-year biochemistry lab, focuses on protein over-expression, purification, and stability of Bacillus circulans xylanase (BCX). BCX is an ideal protein to study as it has important industrial applications and is highly over-expressed, easily purified in a single chromatographic step, and stable. The discovery-based aspect of the project monitors, by circular dichroism spectroscopy, the secondary structure of BCX under conditions that compromise its stabilizing intramolecular forces. Results are written up as a formal lab report in the format of an article in Biochemistry. This module aids in student learning for several reasons: (i) by engaging students in multiple techniques in the context of a single protein, they understand how each technique contributes to the overall production and characterization of BCX, (ii) by allowing students to design a CD-based module, they are required to explore more deeply the forces responsible for protein stability, and (iii) by combining molecular biology, biochemistry, and biophysical chemistry, students more easily see connections between the disciplines.
