HW 3 Fall, 2009 - 1 of 3 Biological Sciences 102 Fall 2009...

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1 of 3 Biological Sciences 102 Fall, 2009 K. Hilt Homework #3 Assigned text problems: Biochemical Calculations (Segel) 4-4 on pp. 223-224 and 4-9 on pp. 237-238 pp. 319 #2, 3, 6, 10, and 12. You need to be able to plot and interpret Lineweaver-Burk graphs. 1. Enzyme Z catalyzes the reaction: A + NAD + B + NADH + H + Besides following the [H + ] using a pH meter, the reaction can be monitored spectroscopically at 340 nm. This is the wavelength at which NADH absorbs light. Some of the first enzymes that were worked on in biochemistry were dehydrogenases, like our enzyme Z, because of this advantage. Radioisotopes were not yet available. a) Diagram what the kinetic assay results would be if the reaction was measured from left to right. What would it be if the assay was measured from right to left? b) Describe how you would accurately determine enzyme Z's K m for NAD + . Describe the experimental set-up and the analysis of data. c) Imagine that twenty I.U.’s of enzyme Z were catalyzing the above reaction for one minute, under V max conditions, in a 3.00 ml assay volume. The assay is buffered with 20 mM
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