Problem Set 3: glycolysis, gluconeogenesis, the pentosephosphate pathway, and the TCA cycle1. A 68 kg adult requires 8000 kJ of energy per day (24 hr) from food. The free energy frommetabolism of food is used to synthesize ATP from ADP and phosphate. Assuming thisconversion goes with 50% efficiency, calculate the weight of phosphate consumed by anadult over 24 hrs. State any assumptions you make (5 points).At 50% efficiency 4000 kJ is available for ATP synthesis.ADP + Pi ’ ATP DG°’ = +30.5 kJ/molTypical cellular concentrations for ADP, ATP and Pi are 140 mM, 1.85 mM and 1.0 mMrespectively (Table 18.2). For non-equilibrium systemsDG’ = DG°’ + RT ln QsoDG’ = 30,500 + RT ln (1.85 x 10-3)/(140 x 10-6)(1.0 x 10-3) = 55 kJ/molIf it requires 55 kJ/mol to synthesize ATP from ADP, we can make 72.8 moles of ATPper day. Assuming physiological pH7, phosphate exists as a 39:61 mixture of NaH2PO4(MW = 120) and Na2HPO4 (MW = 142), an average molecular weight of 133.4. So weturn over about 9.7 kg of phosphate per day.12. In the conversion of glucose to acetyl coenzyme A via glycolysis, which carbon atoms ofglucose are lost as CO2? Justify your answer by drawing the structures of the metabolicintermediates and labeling the carbon atoms (10 points).CHOOHHHHOOHHOHHCH2OHCHOOHHHHOOHHOHHCH2OPCH2OHOHHOOHHOHHCH2OPCH2OOHHOOHHOHHCH2OPPCH2OOCH2OHCHOOHHCH2OPP124356124356CH2OOHCHOCHOOHHCH2OPP124356HCH2OOHCO2P1,62,53,4HPCH2OOHCO2-P1,62,53,4HCH2OHOCO2-1,62,53,4HPCH2OCO2-1,62,53,4PCH2OCO2-1,62,53,4PCH3OCO2-1,62,53,4CH3OHthiamine1,62,5HCH3OS-CoA1,62,5CO23,423. Consider the ten steps of glycolysis, starting with glucose. What would be the effect onpyruvate concentration (increase, decrease or none) of increasing the concentration ofthe following? Give a brief (one sentence) explanation for your answer (10 points).a) ATPATP inhibits PFK, inhibits pyruvate kinase, decreases pyruvate concentrationb) AMPAMP activates PFK, activates pyruvate kinase, increases pyruvate concentrationc) citratecitrate inhibits PFK, decreases pyruvate concentrationd) fructose-1,6-bisphosphatefructose-1,6-bisphosphate activates pyruvate kinase and provides more substrate foraldolase, increases pyruvate concentratione) fructose-2,6-bisphosphatefructose-2,6-bisphosphate activates PFK, increases pyruvate concentration34. A cell undergoing aerobic metabolism is fed with glucose labeled with 14C at carbon 6.Assuming that this is the sole source of carbon for the TCA cycle, what percentage ofCO2 produced by the cell would be labeled with 14C after two turns of the TCA cycle?Assume that the cycle is primed with unlabeled oxaloacetate (10 points).No CO2 is labeled after 2 turns.Note that the filled circles represent half a label – the pyruvate methyl group from C6 ofglucose is labeled, but the second pyruvate methyl group (from C1 of glucose) is not.CO2HOCO2HCO2HHOCO2HCO2HCO2HCO2HCO2HHOCO2HCO2HCO2HOCH2SCoACO2HOCO2HHOCO2HCO2HCO2HCO2HCO2HCH2CO2HCO2HOSCoAOCH3CO2CO2glucoseCO2CO2HOCO2HCO2HHOCO2HCO2HCO2HCO2HCO2HHOCO2HCO2HCO2HOSCoACO2HOCO2HHOCO2HCO2HCO2HCO2HCO2HCO2HCO2HOSCoAOCH3CO2CO2glucoseCO21/41/41/41/41/41/41/41/445. If succinic acid is added to a cell undergoing aerobic metabolism, glycolysis momentarilyincreases to provide more acetyl CoA for the increased oxaloacetate, then decreases asthe increased citrate inhibits PFK. What will happen to glycolysis (increase, decrease, orstay the same) if glutaric acid is added instead of succinic acid? Assume that glutarate isa substrate for succinate dehydrogenase and that trans-glutaconic acid is a substrate forfumarase. It will also help you to know that mitochondrial 2-hydroxyglutaratedehydrogenase is an NAD+ dependent enzyme (10 points).The last 3 steps of the TCA cycle convert succinate to oxaloacetate as follows:Succinate fumarate malate oxaloacetateThe same chemistry converts glutarate to a-ketoglutarate:Glutarate glutaraconic acid 2-hydroxyglutarate a-ketoglutarateSo glutarate gives 1 FADH2 in the succinate dehydrogenase step, 1 NADH in the 2-hydroxyglutarate dehydrogenase reaction, then skips the citrate synthase, aconitase,and isocitrate dehydrogenase steps and goes directly to a-ketoglutarate. From there it isconverted to succinate (1 NADH is produced) and then does a “normal” TCA cycletaking an acetyl-CoA from glycolysis.If aerobic metabolism places a constant demand for NADH on TCA and glycolysis, thisdemand can be met with less glycolysis if glutarate is added.CO2HCO2HCO2HHO2CCO2HCO2HHOCO2HCO2HOCO2HCH2CO2HCO2HCHHO2CCO2HCH2CO2HHOCO2HCH2CO2HO566. Considering glycolysis and the glyoxylate cycle, how many molecules of NADH can beproduced per molecule of glucose? (10 points)Glycolysis gives 2 NADH per glucose at the G3P to BPG step, and there are 2 NADHper glucose produced at the pyruvate dehydrogenase bridging step. The glyoxylatepathway short-circuits the 2 NADH-producing steps between isocitrate and succinate,but still produces 1 NADH per malate at the malate dehydrogenase step. 2 malates areproduced per turn of the glyoxylate cycle, so 2 NADHs are produced. Note that thisrequires both molecules of acetyl-CoA from 1 glucose.Total 6 NADH per glucose (compared to 10 NADH from glycolysis + TCA).7.A bacterium is discovered that doesn’t have a copy of thephosphoglucoisomerase gene. G6P is not converted to F6P in the glycolyticpathway, rather the bacterium uses the pentose phosphate pathway to convertG6P to F6P. Draw the new pathway(s) to F6P.1. ATP; hexokinase/glucokinase2. NADP+; glucose-6-phosphate dehydrogenase3. gluconolactonase4. NADP+; 6-phosphogluconate dehydrogenase5. phosphopentose isomerase6. phosphopentose epimerase7. transketolase8. transaldolase9. transketolase10. ATP; phosphofructokinase11. fructose-1,6-bisphosphate aldolase12. triose phosphate isomerase13. NAD+; glyceraldehyde-3-phosphate dehydrogenase14. ADP; phosphoglycerate kinase15. phosphoglycerate mutase16. enolase17. ADP; pyruvate kinaseglu6Pglucoseglc6Prbu5PR5Pxlu5Psdu7PG3PE4PF6PF6PG3PF1,6BPDHAPG3P1,3BPG3PG2PGPEPpyruvate12,3456789101112131415161778. Reciprocal regulation is a key feature of metabolic pathways that must be able tooperate in both forward and reverse directions. Which enzymes are reciprocallyregulated in glycolysis and gluconeogenesis? Identify the activators andinhibitors for each (5 points).Reciprocal regulation describes the phenomenon whereby the same effector moleculeactivates a particular step in a catabolic pathway