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 Vmax and kcat values for the enzyme trypsin. You will also determine how soybean trypsin inhibitor affects trypsin Vmax and kcat 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 R2 values
  2. Day 1 V vs. [E], with linear curve fit and R2 value
  3. Day 2 Abs vs. time for no inhibitor, all 7 cuvettes on one graph, with linear curve fits and R2 values
  4. Day 2 Abs vs. time with inhibitor, all 7 cuvettes on one graph, with linear curve fits and R2 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 R2 values
  7. Calculations for Vmax and kcat for both plus and minus inhibitor, with appropriate units
  8. A written statement of how the inhibitor affected kcat, and what that tells you about where the inhibitor binds to the enzyme

Objectives