Preparation, Purification, and Secondary Structure Determination of Bacillus circulans Xylanase. A Modular Laboratory Incorporating Aspects of Molecular Biology, Biochemistry, and Biophysical Chemistry

Sal Russo and Lisa Gentile
Department of Chemistry, Western Washington University, Bellingham, WA 98225-9150
J. Chem. Educ., 2006, 83 (12), p 1850
DOI: 10.1021/ed083p1850
Publication Date (Web): December 1, 2006
History, Education, and Documentation

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 Undergraduate

Keywords (Domain):

Biochemistry

Keywords (Pedagogy):

Hands-On Learning / Manipulatives

Keywords (Subject):

Biophysical Chemistry
View: Hi-Res PDF | PDF w/ Links

Citing Articles

View all 3 citing articles

Citation data is made available by participants in CrossRef's Cited-by Linking service. For a more comprehensive list of citations to this article, users are encouraged to perform a search in SciFinder.

This article has been cited by 3 ACS Journal articles (3 most recent appear below).

Tools

SciFinder Links

SciFinder subscribers: Click to sign in | Not a SciFinder subscriber? Learn more at www.cas.org

Explore by:

History

  • Received: August 03, 2009

Recommend & Share

  • Share on ACS NetworkACS Network
  • Add to FacebookFacebook
  • Tweet ThisTweet This
  • Add to CiteULikeCiteULike
  • Add to NewsvineNewsvine
  • Digg ThisDigg This
  • Add to DeliciousDelicious

Related Content