For this week's assignment you will process your data in an Excel spreadsheet, write one sentence of the Discussion section related to this work, provide a literature  Reference for your literature K_m value, and submit your data to the class data spreadsheet. The detailed requirements follow.

Excel Spreadsheet

Note: You now have enough experience with kinetics that the correctness of your calculations will be worth more points than in past assignments. In the instructions below, you will be asked to highlight the cells containing specific sample calculations by filling the cell in yellow. The purpose of this is to help the grader find the required information. Neglecting to do this will result in points deductions.

• Create plots of A₄₀₀ vs. time for all of your raw kinetics data from Part C and graph linear trendlines on each data set to illustrate the quality of the data. You are not required to label the axes.
• Clearly label your substrate concentration calculations (the final concentrations in the assay) and highlight the cell of one sample calculation in yellow.
• Clearly label the slope calculations and highlight the cell of one sample calculation in yellow.
• Clearly label the relative activity calculations and highlight one sample calculation in yellow.
• Create a table that summarizes your [catechol] and v_o (relative activity) data.
• Clearly label the values for your Eadie-Hofstee plot and highlight one sample calculation for the x-axis calculation in yellow.
• Create an Eadie-Hofstee plot and graph a linear trendline of the data.
• In the LINEST array of your Eadie-Hofstee plot data, the four cells for slope, slope standard error, y-intercept, and y-intercept standard error must be highlighted in yellow.
• Clearly label the cells containing calculations and error propagation for V_max/K_m and highlight these cells in yellow.
• Create an organized summary table of your estimates for V_max, V_max/K_m, and K_m with estimated uncertainty. Format this table with the appropriate significant figures and units.

Word Document

Literature comparison

• Use the "Functional Parameters" category in BRENDA to locate a literature K_m value for catechol with Agaricus bisporus tyrosinase. Select a value that has given conditions.
• In one sentence, state this literature K_m value for catechol with Agaricus bisporus tyrosinase and the conditions (pH, temperature, etc.) of the measurement.
• In lab next week, we will discuss how to appropriately compare this value to your measured value.

References

• Using the appropriate format, provide a citation for the primary literature for your K_m value. Note that BRENDA is secondary literature, cite the paper that BRENDA cited, not the database itself.

Submit by next lab meeting

• Your Excel spreadsheet to the online Submissions folder. Highlight the cells for each requested calculation in yellow to make it easier to grade. 

• A digital copy of the Lab 5 Assignment Word document to the Submissions folder.

Post your data online

• Post your groups's V_max, K_m, and V_max/K_m results to your section's data spreadsheet.