2}-0 4.207), B.Tech / Even Q.No. BT 402/ 087 (6-17) Res 2016-17 BIOCHEMISTRY BT 402 Full Marks : 70 Time : 3 hours The figures in the margin indicate full marks. Answer any five questions. 1. With reference to glycolysis and gluconeogenesis, answer the following questions : (@ The irreversible reactions of glycolysis along with their enzymes. (6) Whatare the bypass steps in gluconeogenesis for these reactions ? 3 (© Write the overall equation for glycolysis and gluconeogenesis. 1 @ The different hexokinase isoenzymes of liver and muscle reflect their different roles of these organs in carbo- hydrate catabolism. Explain. 3 @ Fructose-2,6 biphosphate is a potent regulator of glycolysis and gluconeogenesis. How ? 4 2. (@) How can germinating seeds convert the carbon of stored lipids into glucose v (6) Although oxygen does not participate directly in the citric acid cycle, the cycle operates only when oxygen is present. Why ? 2 G/3-120 [ Turn Over ](©) Malate dehydrogenase is found in both the cytosol and in mitochondria. Why ? 2 (@) Ifa molecule of glucose produces two molecules of ATP by substrate level phosphorylation of ADP in glycolysis and the resulting two molecules of pyruvate can each yield 15 molecules of ATP when O, is available, how many glucose molecules will be necessary to produce 160 molecules of ATP by yeast grown under (i) aerobic (ii) anaerobic condi- tions 3 (@) How many molecules of CO, would be evolved in pro- ducing 160 molecules of ATP during growth (i) aerobic (ii) anaerobic conditions ? 3 () How many high energy phosphates are generated in converting 1 mole of glucose to lactate ? 1 . (a) Alanine and glutamine are present in much higher concentration than other amino acids. Why ? 2+2 (6) What causes the deposition of uric acid crystals and describe the biochemical basis for its treatment ? 4 (©) How does the TCA cycle operate if oxaloacetate and a-ketoglutarate are removed for amino acid synthesis ? 2 @ Im animal tissues, the rate of conversion of pyruvate to acetyl-CoA is regulated by the PDH complex. Determine what happens to the rate of this reaction when PDH complex is treated with (a) pyruvate dehydrogenase kinase, ATP, and NADH (b) pyruvate dehydrogenase phosphatase and Ca. 4 . (@) Citrate plays a central role in diverting cellular metabo- lism from the consumption of metabolic fuel to the storage of fuel as fatty acid. Explain. 3 G/13-120 [Continued ] C34) (0) What is the source of NADPH in fatty acid biosyn- thesis ? 2 (c) Why does the concentration of ketone bodies in the blood increase during prolonged starvation ? 3 (d) Describe the carnitine shuttle. 4 (e) How many turns of the fatty acid oxidation cycle are required for the complete oxidation of arachidonic acid to acetyl CoA (20:4) ? a (f) Write the overall equation for palitoyl CoA synthesis. a 5. (a) Isolated spinach chloroplast evolves O, when illumi- nated in presence of Hill’s reagent. Is NADPH produced in this process ? Explain. 2 (6) When a suspension of green algae is illuminated in the absence of CO, and then incubated with “ CO, in the dark, 4 CO, is converted to “ C glucose for a brief time. What is the significance of this observation with regard to the CO, assimilation process and how is it re’.ted to the light reactions of photosynthesis ? 4 (c) What effect on the cell might result from © defect in sedoheptulose 1,7-bisphosphatase in (a) a human hepatocyte and (b) the leaf cell of a green plan’ ? 2 (d) Sucrose synthesis occurs in the cytosol starch synthesis in the chloroplast stroma, yet the two processes are intricately balanced. What factors shift ili reactions in favour of (a) starch synthesis and (b) sucrose synthesis ? 2+2 (e) State the function of the Pi-triose phosphate antiport system. 2 G/13-120 { Turn Over ](ram) 6. (@) Thephosphorylation of glucose to glucose 6-phosphate is the initial step in the catabolism of glucose. The direct phosphorylation of glucose by Pi is described by the equa- tion: Glucose + Pi ->Glucose-6-phosphate + HO 4G” =18.8 KJ/mol. @ Calculate the equilibrium constant for the above reaction. In the rat hepatocyte the physiological con- centrations of glucose and Pi are maintained at ap- proximately 4.8 mM. What is the equilibrium concen- tration of glucose 6-phosphate obtained by the direct phosphorylation of glucose by Pi? Does this reaction represent a reasonable metabolic step for the catabo- lism of glucose ? Explain. 3 Gi) The phosphorylation of glucose in the cell is cou- pled to the gydrolysis of ATP; that is, part of the free energy of ATP hydrolysis is used to phosphorylate glu- cose: Glucose + Pi -> Glucose-6-phosphate + H,O AG® = 13.8 KJ/mol ATP +H,O— ADP + Pi AG” =-30.5 KJ/mol Glucose + ATP — glucose -6-phosphate + ADP Calculate Keq for the overall reaction. For the ATP-depen- dent phosphorylation of glucose, what concentration of glu- cose is needed to achieve a 250 1M intracellular concen- tration of glucose 6-phosphate when the concentrations of ATP and ADP are 3.38 mM and 1.32 mM, respectively ? Does this coupling process provide a feasible route, at least in principle, for the phosphorylation of glucose in the cell ? Explain. 3 (6) Synthesis of activated form of acetate is carried out in an ATP dependant process Acetate + CoA + ATP Acetyl CoA + AMP = PPi G/13-120 [ Continued ] (Ci) The AG® for hydrolysis of acetyl CoA to acetate and CoA is -82.2 KJ/mol and that for hydrplysis of ATP to AMP and PPi is -30.5 KJ/Mol. Calculate the AG” for the ATP de- pendant synthesis of acetyl CoA. 2 (©) Calculate the actual physiological AG for the reaction Phosphocreatine + ADP ->Creatine + ATP Phosphocreatine* + HO — creatine + P?- AG? =—43.0 kJ/mol At 37°C as it occurs in the cytosol of neurons with phosphocreatine at 4.7 nm, creatine at 1.0nm, ADP at 0.73 nm and ATP 2.6 nm. 2 (d) Consider the following interconversion, which occurs in glycolysis : Fructose 6-phosphate — glucose 6-phosphate KR, =1.97 (a) What is AG’ for the reaction at 25°C ? 2 (b) If the concentration of fructose 6-phosphate is adjusted to 1.5 M and that of glucose 6-phosphate is adjusted to 0.50 M, what is AG? 2 G/3-120