Association for Biology Laboratory Education

A rich independent-project-based enzymology lab using plant mitochondrial succinate dehydrogenase
    

Jonathan E. Moore, Ross N. Pringle, & David Becker

Advances in Biology Laboratory Education, 2023, Volume 43

https://doi.org/10.37590/able.v43.art13

Abstract

For well over twenty years, we have successfully run a half-semester inquiry-based enzymology lab at the introductory/ intermediate level. In successive weeks, students use black-eyed pea (Vigna unguiculata) plants to make a crude mitochondrial purification, hone their pipetting skills with a Bradford protein concentration assay, construct a Lineweaver-Burk plot to determine Km and Vmax of succinate dehydrogenase (complex II) in their experimental conditions, consult with their instructor about their independent project proposal, execute their plan, and then write up their results. The literature on SDH is quite diverse, and accordingly the independent projects are quite diverse in nature. Along the way, they learn about differential centrifugation, accurate pipetting, spectroscopy, systematic errors, outliers, and mining the literature. This lab could be easily scaled back to a two-week lab with simpler learning goals.

Keywords:  independent project, enzymology, organelle isolation, differential centrifugation

University of Victoria (2022)