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Exam03_KEY_v2 - MCB 120L Wednesday 11 March 2009 Exam 3 KEY...

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MCB 120L KEY Wednesday 11 March 2009 Circle Instructor: Lindsay Morand Rose Exam 3 Inosine monophosphate dehydrogenase (IMPDH) catalyzes the first step in the de novo biosynthetic pathway ultimately leading to theobromine and caffeine by converting inosine monophosphate (IMP) to xanthosine monophosphate (XMP) with the concomitant reduction of NAD + to NADH. IMPDH + NAD + NADH + IMP XMP Ambitious students of MCB 120L purified IMPDH from 20 g seeds of Theobroma cacao in order to investigate its kinetic and regulatory properties. The protocol was similar to Exp 4 and the result of their efforts is presented in the purification summary table below. sample volume dA 340nm /min Total IU % Yield Total mg Specific activity Fold- purification crude extact 100 ml 0.092 at (1/2) 2 dil 2151 100 200 10.8 1.0 60% pellet 1.0 ml 0.122 at (1/2) 8 dil 1826 85 32.1 56.9 5.3 G-75 2.5 ml 0.069 at (1/2) 7 dil 1290 60 16.0 81 IU/mg 7.5 AMP-affinity 2.0 ml 0.070 at (1/2) 7 dil 1032 48 2.0 516 48 1. The IMPDH assays were performed in the microplates similar to the LDH assays, except saturating
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