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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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Abstract
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.
Keywords (Audience):
Upper-Division UndergraduateKeywords (Domain):
BiochemistryKeywords (Pedagogy):
Hands-On Learning / ManipulativesKeywords (Subject):
Biophysical ChemistryCiting Articles
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This article has been cited by 4 ACS Journal articles (4 most recent appear below).
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Nancy McKenzie, James McNulty, David McLeod, Meghan McFadden, and Naresh BalachandranJournal of Chemical Education2012 89 (6), 743-749A new undergraduate program in chemical biology was launched in 2008 to provide a unique learning experience for those students interested in this interdisciplinary science. An innovative undergraduate chemical biology laboratory course at the third-year ...
Preparative Protein Production from Inclusion Bodies and Crystallization: A Seven-Week Biochemistry Sequence
Megan J. Peterson, W. Kalani Snyder, Shelley Westerman, and Benjamin J. McFarlandJournal of Chemical Education2011 88 (7), 986-989Preparative Protein Production from Inclusion Bodies and Crystallization: A Seven-Week Biochemistry Sequence
Megan J. Peterson, W. Kalani Snyder, Shelley Westerman, and Benjamin J. McFarlandJournal of Chemical Education2011 88 (7), 986-989We describe how to produce and purify proteins from Escherichia coli inclusion bodies by adapting versatile, preparative-scale techniques to the undergraduate laboratory schedule. This 7-week sequence of experiments fits into an annual cycle of research ...
Circular Dichroism Spectroscopy in the Undergraduate Curriculum
Adam R. UrbachJournal of Chemical Education2010 87 (9), 891-893Circular Dichroism Spectroscopy in the Undergraduate Curriculum
Adam R. UrbachJournal of Chemical Education2010 87 (9), 891-893Circular dichroism spectropolarimetry (CD) is a method of optical spectroscopy that seems in most practical ways like UV−visible spectroscopy. The main difference between the two methods is that CD, instead of measuring the absorbance of light as a ...
Teaching Protein Purification and Characterization Techniques. A Student-Initiated, Project-Oriented Biochemistry Laboratory Course
Gina MacDonaldJournal of Chemical Education2008 85 (9), 1250Teaching Protein Purification and Characterization Techniques. A Student-Initiated, Project-Oriented Biochemistry Laboratory Course
Gina MacDonaldJournal of Chemical Education2008 85 (9), 1250This report describes a biochemistry laboratory that is completely project-oriented. Upper-level biology and chemistry majors work in teams to purify a protein of their choice. After the student groups have completed literature searches, ordered reagents, ...
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- Received: August 03, 2009
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