A new inquiry-based teaching laboratory, suitable for both Physical Chemistry and Biochemistry Labs, is presented in an effort to elucidate the concepts of Michaelis-Menten kinetics. The lab is focused on the development of procedures by the students, using the existing literature as a guide. In Part I of the lab, students determine the Michaelis-Menten kinetic parameters of a model enzyme, papain, using regression techniques of their own choice, such as Lineweaver-Burk, Hanes-Woolf, Eadie-Hofstead, Eisenthal's Direct Line, Cornish-Bowden, or nonlinear techniques. Papain was chosen as a model enzyme due to its low cost and ease of rate determination using colorimetric methods. In Part II of the lab, students consult the literature to design their own experiment in order to examine an inhibitor of their choice from those available in the lab. Typical inhibitors used are targeted class inhibitors (E-64), alkylating agents (iodoacetamide), and denaturants (concentrated HCl, ethanol, urea). The students are asked to determine the type of inhibition, by determining if the inhibition is reversible or irreversible, and if reversible, whether the inhibitor is competitive, noncompetitive, uncompetitive, or a mixture of types. Depending upon the type of inhibition, students then quantify the appropriate kinetic and inhibition parameters. Students are required to prepare a laboratory report in which all procedures must be explained and defended, as if they were to publish their results in an academic journal. The focus of the lab is on the preparation of students for careers in both upper academia and industry through the development of independent thinking skills as facilitated by an inquiry-based approach. The concentration of the lab material on the kinetics of a model enzyme is also beneficial to those students interested in medical and biochemical careers.<br/>

Inquiry-based lab for the investigation of Michaelis-Menten kinetics

251st National Meeting (2016)

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