Question

Question 3 10 points Save Ans It is important that you understand how to use the data generated by an absorbance based enzyme assay and relate this back to the enzyme activity Using the steps provided below. calculate the actvity of GPOH in the original preparation expressed as units ml tt two decimal places) under the following conditions: 75 uL glucose 6-phosphate dehydrogenase (G6PDH) was assayed in a reaction mixture with a total volume of 1.20 mL. The reaction was followed by measuring the tate of appearance of NADH from the change in Absorbance at 340 nm. The initial rate of change of Absorbance (AA) was 0.55 A min Note: The extinction coefficient at 340 nm ( εμ m) of NADH is& 220 M 1cm while for NAD. E 340 nm, is O M , cm', Use the following steps . calculate Δ(NADHI man, using the Beer-Lambert equation (A . calculate the total mole of NADH appearing per min from the total volume of the reaction mixture and convert the value to mmol min . convert mmol min to units (see definition of activity in the introduction) divide by mL enzyme (G6PDH) added to the reaction mixture

Answer 1

M(NADH)min? Nouw, Gnvert the value tor mmel wmin An El n - 26.umol i264 0Ds mmolmin timm A- Ec Arii26 yumol min" 26-4 U 0.55 min 6220Mm per ml 0.03s rmix Given: Yotal volume .20ml -1 tn v Velume →26.40 0.07 ml 0.088n(NADH) 30D