A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics ?alkaline phosphatase as a model

MIQUET JG ; SOTELO AI ; GONZALEZ L ; GONZALEZ LEBRERO RM

Biochemistry and Molecular Biology Education

John Wiley & Sons, Inc.

Año: 2019

Enzyme kinetics is an essential topic inundergraduate Biochemistry courses. A laboratory work that covers the principalbasic concepts of enzyme kinetics in steady state is presented. The alkalinephosphatase catalyzed reaction of phenyl-phosphate hydrolysis was studied as amodel. The laboratory experience was designed to reinforce the concepts ofinitial velocity dependence on substrate and enzyme concentration, and tohighlight the importance of the accurate determination of initial reactionrate. The laboratory work consists in two parts, in which students firstdetermine the enzyme concentration and the time to be used in the followingsession to obtain the kinetic parameters (KM and Vmax) bynon-lineal fitting of the Michaelis-Menten equation to the initial velocitydependence with substrate concentration results. The experimental methodologyis robust, the cost per student is low and the equipment and reagents used areof easy access.