Title: Milestone Lab: Trypsin Kinetics
Course: BTEC-2020-301-302-303-F21 > Section: Enzyme Kinetics

Module4 Milestone Lab: Trypsin Kinetics

Instructions

Note: You must complete this lab before continuing on in the course.

The Trypsin Kinetics Protocol will guide you through the process of determining the V_max and k_cat values for the enzyme trypsin. You will also determine how soybean trypsin inhibitor affects trypsin V_max and k_cat values.  Read and follow the Protocol.

Mod 4 Enzyme Kinetics Trypsin protocol rev Nov 2020.pdf

Upon completion of the lab, upload a document that includes the following graphs, all well labeled (including units!), and other items.

1. Day 1 Abs vs. time, all 7 cuvettes on one graph, with linear curve fits and R^2 values
2. Day 1 V vs. [E], with linear curve fit and R^2 value
3. Day 2 Abs vs. time for no inhibitor, all 7 cuvettes on one graph, with linear curve fits and R^2 values
4. Day 2 Abs vs. time with inhibitor, all 7 cuvettes on one graph, with linear curve fits and R^2 values
5. Day 2 V vs. [S], both plus and minus inhibitor on one graph so they can be compared (do not fit a curve, as they should be hyperbolic and Excel can’t fit that)
6. Day 2 1/V vs. 1/[S], both plus and minus inhibitor on one graph so they can be compared, with linear curve fits and R^2 values
7. Calculations for V_max and k_cat for both plus and minus inhibitor, with appropriate units
8. A written statement of how the inhibitor affected k_cat, and what that tells you about where the inhibitor binds to the enzyme

Objectives

- Measure an enzyme’s rate of reaction with various substrate concentrations.
- Create a Michaelis-Menten plot.