Carbohydrate Metabolism INTRODUCTION Having understood the form and properties of carbohydrates, it is time to see how carbohydrates are used to perform their primary function — generate energy. In order to use the energy stored in carbohydrates, they need to be bro- ken down. Thus, in this chapter we shall study the metabolism of carbohydrates In addition, we shall also see how carbohydrates may undergo various chemical alterations in order to synthesise other important biomolecules. In understanding carbohydrate metabolism, one should know that glucose is the ultimate carbohydrate. Most metabolic reactions in the body are aimed at converting the carbohydrate into glucose, so that it can be easily used. Other forms, such as fructose and galactose, are also used in some pathways. However, these constitute a very small proportion of our dietary consump- tion of carbohydrates is in the form of disaccharides and polysaccharides. The journey from these forms to the monosaccharide form, and the process- ing of that monosaccharide form, is what constitutes the metabolism of carbohydrates. Intertissue balance in the utilisation and storage of carbohydrates (glucose) is accomplished mainly by the action of hormones. Thus, the metabolism of carbohy- drates involves all these processes. This chapter includes the diseases involved thereof. Each pathway is described by elaborating location; flow sheet diagram; salient features such as rate limiting steps, modulations; regulations; energetic and functions of the pathway. Q1. Explain digestion and absorption of carbohydrates. OR Cellulose cannot be digested by the human body. Explain why. OR Carbohydrates cannot be digested in the stomach. Explain why. OR Why does digestion begin in the small intestine? OR 36Chapter 4 — Carbohydrate Metal Explain the role of chloride ions and pH in the mouth with respect to digestion. Ans. * The sites of digestion and absorption in human beings are shown in Fig. 4.1. * Glycogen and starch are the major dietary polysaccharides. Few monosaccha- rides are also present in the diet. © Digestion of carbohydrates occurs largely in the mouth and the intestinal lumen. * The salivary amylase acts briefly on dietary starch in the mouth. This is an a-amylase which attacks the a (14) bonds. * Cellulose, which has 8 (14) glycosidic linkages, cannot be digested by the human body because a-amylase digests only a (14) bonds. Cellulose, there- fore, acts as a dietary fibre. © The a-amylase does not attack « (1-6) bonds. © The products after action of a-amylase are as follows: © Maltose * Maltotriose © Limit dextrins made up of highly branched structures * Digestion of carbohydrate cannot take place in the stomach because the high acidity of the gastric hydrochloric acid inactivates the salivary a-amylase. Starch Lactose oan ‘Sucrose Fructose Salivary e-amylase Cl, pH6.6-6.8 ¥ STOMACH HCI (destroys amylase) ¥ Giycogen, starch maltose, limit dextrins, ) > | INTESTINE. lactose, sucrose, fructose Pancreatic amylase, pH 7.1 ¥ > | EPITHELIAL CELLS | (Brush border) Maltose, limit dextrins, lactose, sucrose fess Disaccharidase, oligosaccharidase; pH 5.0-7.0 Limit dextrinase Portal vein ¥ Transport Glucose, galactose, > | LIVER fructose Galactose, fructose metabolism v Glucose > CIRCULATION Cellulose ¥ FAECES, Fig. 4.1 Digestion and absorption of carbohydrates.38 Medical Biochemistry: Preparatory Manual for Undergraduates * When the contents of the stomach reach the small intestine, the acidity of the contents is neutralised by the action of bicarbonate secreted by the pancreas. ; © Pancreatic amylase, which is an « (1-4) glucosidase, continues to digest the starch in the intestines. © The disaccharidases and oligosaccharidases act to release the monosaccha- rides, in the upper jejunum. © The bulk of the dietary sugars are absorbed by the duodenum and jejunum. © By different transport mechanisms, the sugars enter the portal circulation and are utilised. Q2. Write a short note on the roles played by amylases. Ans. * The salivary amylase acts briefly on dietary starch in the mouth. This is an a- amylase which attacks the a (1—>4) bonds. * Maltose * Maltotriose * Limit dextrins made up of highly branched structures Lo Starch © Digestion of carbohydrates cannot take place in the stomach because the high acidity of the gastric hydro- Peper Ho: chloric acid inactivates the salivary TTT a-amylase. Salivary and * When the contents of the stomach pancreatic a-amylase reach the small intestine, the acidity of the contents is neutralised by the ee action of bicarbonate secreted by q(14) the pancreas. * Pancreatic amylase, which is an a Bekins eens Isomaltose (1-4) glucosidase, continues to di- Tisaecharides gest the starch in the intestines. aaa © The amylase shows no activity for « Dent (136) bonds (Fig. 4.2). a. Fig. 4.2 Action of salivary and pancreatic Q'8. Give a diagrammatic represen- %-amylase. tation of the fate of glucose. Ans. * Glucose isa versatile molecule. Itis used not only to produce energy, but is also used to synthesise a number of other products. * Glucose plays a central role in the anabolic as well as catabolic pathways. The pathways of glucose metabolism are represented in Fig. 4.3. QA. Write a note on the different modes of glucose transport. OR Write a short note on glucose transporters. OR Explain briefly the tissue specificity of glucose transporters in facilitated diffusion. Ans. Modes of glucose transportGlucose ————> Sorbitol ————+ Fructose Chapter 4 — Carbohydrate Metabolis Glycogenolysis (Glucose-6-P) Glucose-1-P ————> alycogen | Glycogenesis HMP’ UDP-Glucose shunt | Glucuronate ——» Mucopolysaccharides Uronic acid pathway Ribose-5-P- NADPH Nucleotides ve A Anaerobic Pyruvate Lactate J glycolysis Acetyl CoA ;}—> Cholesterol —> Steroids Ss Fatty acids —> Triglycerides, phospholipids y TCA Ketone bodies ee acids HO Co, Fig. 4.3 Fate of glucose. * Glucose is transported through the cells of the intestine by binding to trans- port proteins (Fig. 4.4). ° There are two types of glucose transport proteins, namely the facilitative glucose transporters and Na*dependent glucose transporters. (i) Facilitated diffusion © There area nu of these; gluco: imber of similar proteins, found in the plasma membrane se transporter GLUT-1 to GLUT-5 is predominant. © They are tissue specific. For example: - GLUT-1 is present in most cells, but primarily in erythrocytes, placenta and brain. G GLUT(1-5) Facilitated glucose transporter GLUT-glucose transporter Nat Gal Na‘ glucose Passive Na” co-transporter diffusion 2K* okt Na*, K*-ATPase pump, ¥ Fig. 4.4 Transport of glucose.| 40 Medical Biochemistry: Preparatory Manual for Undergraduates Glucose transporters Sites Nat-glucose co-transport SGLTt Small intestine, renal tubules sGLT2 Renal tubules Facilitated diffusion | @uuTi Placenta, blood-brain barrier, red cells, many organs GLUT 2 8-cells of islets, liver, epithelial cells of small intestine, kidneys | GLUT3 Brain, placenta, kidneys many other organs GLUT 4 Skeletal and cardiac muscle, adipose tissue, other | tissues | Fructose transport Jejunum, sperm GLUT 5: GLUT, glucose transporter; SGLT, sodium-dependent glucose transporters. — GLUT 3 exists in the neurons, placenta and testes. GLUT-3 supple- ments the action of GLUT-1 when energy demand is high. The glucose attaches to one of these transporter proteins, which then by a conformational change carries the glucose into the cell. This move- ment via a glucose transporter is stimulated by insulin into the cells of muscle and adipose tissue. — GLUT is present in insulin-targeted tissues such as adipose tissue, cardiac and skeletal muscle. © No energy is required for this process. * Additional GLUT-6 to GLUT-12 have been identified but their roles are not yet deciphered. The details of various glucose transporters are given in Table 4.1. (ii) Na*-dependent transport © The transport of glucose from region of low concentration to that of high concentration is promoted by the co-transport of Na*. © Na*-K*-ATPase pump. The sodium ions are actively removed to the extracellular fluid via the Na* K*-ATPase pump. © This forms the basis for the oral rehydration therapy (ORS). ORS contains salt and sugar. The glucose gets transported into Na* ions, relieving the person of dehydration. (iii) Passive diffusion * The process is indirectly energy dependent. Glucose can also move by passive diffusion into the cells. Q5. Explain Embden-Meyerhof-Parnas pathway. OR What is glycolysis? Explain aerobic and anaerobic glycolysis along with the energetics, OR Explain the role of K,, using glucokinase and hexokinase as examples.Aerobic Pyruvate asia Anaerobic oe glycolysis Lactate Cytoplasm Mitochondria Fig. 4.5 Compartmentalisation of glycolysis. OR Write a note on the irreversible steps in the glycolytic pathway. Ans. * Once inside a cell, the chief function of the glucose molecule is to release the energy stored within it. However, before it can do this, it has to undergo a gruelling series of changes. Glycolysis is the process of breakdown of glucose to pyruvate or lactate. It is also called Embden-Meyerhof-Parnas (EMP) pathway location. Compartmentalisation. Pyruvate and lactate are formed in the cytosol (Fig. 4.5), whereas the oxidation of acetyl CoA takes place in the mitochondrion. There are two phases of glycolysis: (i) Preparatory phase. During this phase, phosphorylation of glucose and its conversion to glyceraldehyde 3-phosphate take place. (ii) Pay-off phase. During this phase conversion of glyceraldehyde 3-phosphate to pyruvate and the coupled formation of ATP takes place. Purpose of giycolysis is to provide fuel in the form of ATP and intermediates for the metabolic pathways. For many tissues, glycolysis is the only energy yielding pathway. Reactions of glycolysis © The reactions of glycolysis can be studied under two phases — preparatory and pay-off phases. Preparatory phase Reactions under preparatory phase (Fig. 4.6) are as follows: ® Glucose is converted into glucose 6-phosphate with the help of enzymes, hexokinase and glucokinase. © Glucokinase acts as a ‘glucostat’ and comes into play when glucose is in the bloodstream. It is present predominantly in the liver. It is specific for glucose and has high Ky, (~10 mM) and is not inhibited by glucose phosphate at physiologic concentrations. It is induced by insulin. Hexokinase is found in most of the tissues and comes into play when blood glucose is low. It has alow K,, (~0.1 mM) and is inhibited by glucose 6-phosphate (product inhibition). This is an irreversible regulatory step. Glucose 6-phosphate is isomerised to fructose 6-phosphate with the help of phosphoglucoisomerase. This is a reversible reaction. Fructose 6:phosphate is phosphorylated by ATP to form fructose 1,6-bisphosphate (FL,6BP). The enzyme used is phosphofructokinase I (PFK I). This is an’ irreversible and regulatory step.42 Medical Biochemistry: Preparatory Manual for Undergraduates Glucose Hexokinase (all calls except liver) ADP (Km~ 0.1 mM) @ Insulin. ———» Glucokinase ©Glucagon —(K,,~10 mM) in liver Mg? Regulatory enzyme (Inhibited by glucose 6-P) a-Glucose 6-phosphate Phosphoglucose isomerase Fructose 6-5 ie Rate-limiting step @ F2,6 BP, AMP, PFK Il @ Insulin irreversible, inducible pesca et (PFKI) © Citrate, ATP © Glucagon ‘A. Most tissues Fructose 1,6-bisphosphate B. Liver and kidney Zn containing Aldolase Dihydroxyacetone phosphate + Glyceraldehyde 3-phosphate eC Triose phosphate isomerase (2) Glyceraldehyde 3-phosphate Fig. 4.6 Preparatory phase of glycolysis. ° PEK I is activated by fructose 6-phosphate, adenosine monophosphate (AMP), PEK Il, by fructose 2,6-bisphosphate (F2,6BP) and insulin. It is inhibited by ATP, citrate and glucagon. Deficiency of this enzyme in skeletal muscle causes fatigue. * F1,6BP is cleaved to form two triose phosphates, namely, dihydroxyacetone phosphate (DHAP) and glyceraldehyde 3-phosphate with the help of the enzyme aldolase. * This is a Zn-containing enzyme. A number of isoenzymic forms of aldolase exist. Aldolase A is present in most tissues whereas aldolase B is present in liver and kidney. * The DHAP is converted with the help of triose phosphate isomerase to glyceraldehyde 3-phosphate. M° Thus, the starting unit of six carbons is broken down into two units of three carbons each. This ends preparatory phase. Pay-off phase Reactions of pay-off phase (Fig. 4.7) are as follo Chapter 4 — Carbohydrate Metabolism _ © Glyceraldehyde 3-phosphate is oxidised to form a high-energy compound 13 bisphosphoglycerate (1,3-BPG) by the enzyme glyceraldehyde $phosphate dehy- drogenase. Phosphorylation occurs at the expense of an inorganic phosphate. ° This is the first of the energy-conserving steps. It requires coenzyme NAD*, the oxidised form of nicotinamide dinucleotide which gets (2) Glyceraldehyde 3-phosphate i+ NAD* Glyceraldehyde 3-phosphate dehydrogenase NADH + Ht (to ETC) - Todoacetate (2) [e:Bisphosphogiyeerate (SLP) ADP Phosphoglycerate ki ASST ee aed Mg? ATP. (2) 3-Phosphoglycerate | Phosphoglycerate mutase (2) 2-Phosphoglycerate i Enole Fluoride: moe H,0 (2) Phosphoenolpyruvate @ Insulin ee aor © Glucagon Irreversible (SLP) Pyruvate kinase estate ATP y (2) Pyruvate _— NADH+HY <~ Lactate dehydrogenase ssi NAD* (2) Lactate Fig. 4.7 Pay-off phase of glycolysis. “PETCa4 Medical Biochemistry: Preparatory Manual for Undergraduates reduced in this step. The reduced NADH + H’* is reoxidised via the elec- tron transport chain where it yields three ATPs per molecule of NADH. © The high-energy phosphate from 1,3-BPG is transferred to ADP via the enzyme phosphoglycerate kinase to yield 3-phosphoglycerate. This reaction is reversible. This is an example of substrate level phosphorylation (SLP). Arsenate uncouples oxidation and phosphorylation at this step. 4 © The enzyme phosphoglycerate mutase shifts the phosphate group o 3-phosphoglycerate to carbon-2 to form 2-phosphoglycerate. * 2Phosphoglycerate gets dehydrated with the help of the enzyme enolase to form a high-energy compound, phosphoenolpyruvate. Fluoride inhibits enolase. This is a reversible reaction. Mg?* or Mn2* ions are required. © The high-energy phosphate from phosphoenolpyruvate is transferred to pyruvate with the help of pyruvate kinase. This is another example of SLP. Itis an irreversible reaction. * It requires K* along with Mg?* as cofactors. This is a regulatory step. Pyru- yate kinase is activated by F1,6-BP, high carbohydrate intake and high insulin levels. It is regulated by covalent modification. Increased levels of cAMP, alanine, fatty acids and acetyl CoA serve as inhibitors of the reaction. This is the last reaction of aerobic glycolysis. d * In anaerobic glycolysis, the conversion of pyruvate to lactate is the essential step. Anaerobic here does not actually mean ‘in the absence of oxygen’. * In this case, it refers to the reoxidation of NADH (generated from the reac tion catalysed by the enzyme glyceraldehyde 3-phosphate dehydrogenase) eS without the requirement of oxygen and, hence, the electron transport chain. o2 ° The reaction is catalysed by lactate dehydrogenase. In exercising skeletal ct muscle, lactate is formed and released into the blood. Lactate dehydroge- : nase occurs in five isoenzymic forms. Details are discussed in the chapter on enzymes. Q6. Why are glycolytic intermediates phosphorylated? Ans. All the glycolytic intermediates are phosphorylated. They perform three functions: © The phosphate groups are ionised at pH 7.0, giving the intermediates a nega- tive charge. Plasma membrane is impermeable to the charged molecules. Therefore, the phosphorylated molecules do not diffuse out. * Phosphate groups conserve the energy. High-energy compounds such as 1,3-BPG and phosphoenolpyruvate (PEP) donate groups to ADP to form ATP. * Binding of phosphate groups to the active sites of enzymes lowers the activation energy. The phosphate groups form complexes with Mg** The binding is specific for these complexes. Most of the reactions of glycolysis require Mg?* ions. QZ7. What are the types of glycolysis? Ans. There are two types of glycolysi * Anaerobic glycolysis occurs when oxygen is scarce. Two ATP molecules are produced and the final product is lactate. Lactate production from glucose Occurs without the need for oxygen to reoxidised the NADH; hence, itis called anaerobic glycolysis. The glycolysis that occurs in exercising skeletal muscle is, an example of anaerobic glycolysis. * Aerobic glycolysis occurs when oxygen is plentiful. Eight ATP molecules are pro- duced during this process and the final product is pyruvate. It occurs in smooth muscles that contract for a long time such as muscles of the gastrointestinal tract.Chapter 4 — Carbohydrate Metabolism Q8. Write a note on the energetics of glycolysis under aerobic and anaerobic conditions with details of the enzyme involved. Ans. * The energy generated is stored in the form of ATP, which is the ultimate energy currency of the cell. © The energy stored in the ATP molecule is released to perform different bodily functions through a mechanism called ETC. This is described in the chapter on ‘biological oxidation’. Tables 4.2 and 4.3 explain the formation and consumption of ATP in aerobic and anaerobic glycolysis, respectively. Under aerobic conditions © Predominant tissues where aerobic glycolysis occurs are muscle, RBCs. brain, gastrointestinal tract, skin, kidney, medulla, retina and heart. Overall reaction: Glucose + 2P; + 2NAD* + 2ADP —>2 Pyruvate + 2ATP + 2NADH +2H* + 2H,O Under anaerobic conditions * Anaerobic glycolysis occurs in erythrocytes, lymphocytes, white blood cells, kidney, medulla, tissues of eyes and skeletal muscles (Table 4.3). Overall reaction: Glucose + 2P; + 2ADP ——> 2-Lactate + 2ATP + 2H;O © Glycolysis is used for the synthesis ofa number of substances; besides provid- ing energy, ribose 5-phosphate is formed from glucose 6-phosphate. Formation and consumption of | Reaction catalysed by ATP formed/consumed Hexokinase/glucokinase Phosphofructokinase Glyceraldehyde 3-phosphate dehydrogenase Phosphoglycerate kinase Pyruvate kinase Formation and Reaction catalysed by ATP formed/consumed Hexokinase Phosphofructokinase Phosphoglycerate kinase Pyruvate kinaseMedical Biochemistry: Preparatory Manual for Undergraduates © Other sugars such as mannose and sialic acids are also formed. Serine is synthesised from 3-phosphoglycerate and alanine from pyruvate, and glyc- erol S-phosphate is formed from dihydroxyacetone phosphate. © 2,3-bisphosphoglycerateis also an important by-product Q9. What are the clinical manifestations of pyruvate kinase deficiency? Ans. Deficiency of pyruvate kinase causes decreased production of ATP. RBCs are affected. The ionic gradient across the membrane cannot be maintained due to lack of ATP. Lysis occurs, leading to haemolytic anaemia. Q.10. Explain the central role played by glucose 6-phosphate in metabolism. Ans. Glucose 6-phosphate is capable of undergoing glycolysis, gluconeogenesis, HMP pathway and glycogen metabolism. Q111. Give examples of substrate level phosphorylation in glycolysis. Ans. Two examples of SLP in glycolysis are as follows: ° The transfer of high-energy phosphate bond to ADP to form ATP from 1,3-bisphosphoglycerate, The enzyme involved is 3-phosphoglycerate kinase and the product is 3-phosphoglycerate. Glyceraldehyde-3-phosphate Phosphoglycerate Clesretemncemmaneecooice kinase 3-phosphoglycerate S-phosphate wae |1,3-bisphosphoglycerate| aN phosphogly i NAD+ NADH +H* ADP ATP ei FI © The high-energy enol phosphate bond present in phosphoenolpyruvate is transferred to ADP to form ATP by pyruvate kinase. This is the second example of SLP in glycolysis. Pyruvate kinase [Phosphoenolpyruvate a” Pyruvate ADP ATP. Q.12. What are the diseases associated with impaired glycolysis? Ans. Some of the diseases associated with impaired glycolysis are as follows: * Hexokinase deficiency: In case of hexokinase deficiency, red blood cells contain less amounts of intermediates like 1,3BPG and 2,3-BPG. 2,3-BPG lowers the affinity of haemoglobin for oxygen, resulting in the release of oxygen in the tissue, Therefore, decrease in the levels of 2,3-BPG reduces the amount of oxygen available in the tissues. * Lactic acidosis: It is the disease in which lactate, the final product of anaerobic glycolysis, accumulates. * In lactic acidosis, the lactate levels are >5 mM which lead to a decrease in blood pH to below 7.2. * It occurs when there is a collapse of the circulatory system, as in myocardial infarction, pulmonary embolism, and when there is a failure to provide enough oxygen to the tissues. © The excess oxygen required to recover from a period of unavailability of oxygen is termed oxygen debt. Lactate levels in blood provide for early detec tion of oxygen debt. These levels are used to measure the presence and sever- ity of shock.Chapter 4 — Carbohydrate Metabolism — Point to note: Exercising muscle: Lactate gets accumulated in the actively exercising muscle and may lead to cramps. Q.13. How is glycolysis regulated? * Hormonal. Insulin stimulates glucokinase, PFK I and pyruvate kinase to favour glycolysis. Glucagon inhibits the same. Availability of substrates. Supply of substrates such as glucose, glucose 1- or 6-phosphate, ADP, P, and NAD* affects glycolysis. The three irreversible reactions catalysed by hexokinase or glucokinase, phospho- fructokinase and pyruvate kinase are the major sites of regulation of the pathway. High level of these enzymes favours the conversion of glucose to pyruvate. Oxidation-reduction. Reactions of glycolysis are dependent on ratios of NAD*: NADH + H¢* and pyruvate: lactate and availability of oxygen. Q.14. Explain why blood for glucose estimation is collected in fluoride containing tubes. Ans. Blood collected for glucose estimation is drawn in fluoride tubes as it inhibits the enzyme enolase. It thus prevents the utilisation of blood glucose facilitating the analysis of blood glucose levels. Q.15. Briefly discuss the relationship between glycolysis and ischaemia. Ans. Ischaemia means limited blood flow. In this condition, the tissue is deprived of oxygen and nutrients. Anaerobic glycolysis is activated and lactic acid production increases. During myocardial infarction, damage occurs during the reoxygenation of the tissue and not during hypoxia. Maintenance of glycolysis plays a special role in protecting membrane function in ischaemia. Q.16. What is the relationship between glycolysis and cancer cells? Ans. In cancer cells, glycolysis occurs very rapidly so that more pyruvate is formed than metabolised. This causes formation of lactate, and hence acidosis occurs locally. This occurs even in the presence of sufficient oxygen and is called the Warburg effect. Q.17. Explain the Rapoport-Luebering cycle. OR Explain how 2,3-BPG acts as a regulator of oxygen transport. Ans. * Rapoport-Luebering cycle occurs in erythrocytes and is also called the bisphosphoglycerate shunt. Itis the pathway for the formation and degradation of 1,3-bisphosphoglycerate (Fig. 4.8). * Ivis a ‘side reaction’ of the glycolytic pathway. * 2,3:-BPG combines with haemoglobin, causing a decrease in affinity of the latter for oxygen and a displacement of oxygen-haemoglobin dissociation curve. 2,3-BPG helps oxyhaemoglobin to unload oxygen. Thus, it acts as a regulator of oxygen transport. HbO, + 2,3-BPG ——> Hb-2,3-BPG + O Oxyhaemoglobin —> DeoxyhaemoglobinMedical Biochemistry: Preparatory Manual for Undergraduates Glyceraldehyde 3-phosphate PB NAD* Glyceraldehyde 3-phosphate dehydrogenase NADH + H* 1,3-Bisphosphoglycerate ADP. Phosphoglycerate Mutase kinase ATP 3-Phosphoglycerate 2,3-Bisphosphoglycerate Utilized: in anaerobic alycolysis 2,3-BPG phosphatase. P, Fig. 4.8 Rapoport-Luebering cycle. * Fetal haemoglobin (HbF) consists of two polypeptide chains called w-globin chain and two y-globin chains (ayo). After birth, during the first few weeks, HDF is replaced by adult haemoglobin (HbA). HbA is made up of two chains and two B chains (a282). The difference in the amino acid composi- tion of the two haemoglobins causes HbF to have a lower affinity for 2,3-BPG than HbA. Consequently, HbF has a greater affinity for oxygen. Thus, oxygen is efficiently transferred across from HbA to HDF, i.e. from mother to fetus. Q)18. Give the reason for hyperventilation in high altitudes. Ans. Ascent to high altitudes, exercise, anaemia and alkalosis increases the concentration of 2,3-BPG, which reduces the oxygen affinity of haemoglobin and promotes the release of oxygen which is required in such conditions. This explains hyperventilation. Q.19. Write a short note on metabolism of fructose. Ans. * Fructose forms a very important part of our diet. It is present in large amounts in fruits, table sugar and sweets. * Fruits contain sucrose, which when acted upon by sucrase, gets hydrolysed to form glucose and fructose. * Fructose enters the epithelial cells by facilitated diffusion. * Fructose can also be synthesised in the body by the sorbitol pathway. It is the source of energy in sperms. Fig. 4.9 represents the metabolism of fructose as it ‘occurs in liver, intestinal mucosa and kidney. * Fructose can be metabolised only after phosphorylation. * Itcan be phosphorylated either by fructokinase or hexokinase. * Phosphorylation by fructokinase is the primary mechanism. AChapter 4 — Carbohydrate Metabolism Fructose ATP Fructokinase [Essential fructosuria ADP Fructose 1-phosphate Aldolase B [Fructose intolerance D-Glyceraldehyde Dihydroxyacetone phosphate ATP Triose kinase ADP. Glyceraldehyde 3-phosphate | Giycolysis Fig. 4.9 Metabolism of fructose. * Fructose I-phosphate is formed in this reaction which is acted upon by an isozyme of aldolase (aldolase B). ° Only when fructose is present in very large amounts, the enzyme hexokinase phosphorylates it further to form F1,6BP. This occurs only on rare occasions. ® Aldolase A is the isozyme of aldolase which cleaves F1,6BP to glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. These intermediates then enter the glycolytic pathway. Aldolase A is present in the muscle. © Though fructose is utilised through the glycolytic pathway, it bypasses the main regulating step of glycolysis, which is catalysed by phosphofructokinase. There- fore, consumption of large amounts of fructose leads to hypertriglyceridaemia. Q.20. Describe metabolic abnormalities related to fructose metabolism. Ans. © Essential fructosuria is a metabolic abnormality due to the deficiency of fructo- kinase, the first enzyme in the fructose metabolic pathway. It is a benign condi- tion because no toxic metabolites of fructose accumulate in the liver and the patient remains asymptomatic. © Hereditary fructose intolerance is due to the absence of aldolase B. It causes severe hypoglycaemia, vomiting and jaundice. Treatment involves removal of fructose and sucrose from the diet. Hepatic failure and death can occur. Q.21. Write briefly about the sorbitol pathway for synthesis of fructose. Ans. © This is also called polyol pathway (Fig. 4.10). It occurs largely in spermatozoa. © Sorbitol is formed from glucose by the action of aldose reductase in many tissues such as lens, retina, placenta, RBCs, ovaries and spermatozoa. © The enzyme aldose reductase is not found in the liver. © In liver, ovaries, sperm and seminal vesicles, sorbitol dehydrogenase converts sorbitol to p-fructose. © Both the enzymes require NADPH + H*. * Fructose metabolism is not affected by insulin.Medical Biochemistry: Preparatory Manual for Undergraduates Absent in liver Mc ductase pote D-Sorbitol (polyol) NADPH + H* NADP* NADPH + Ht Sorbitol dehydrogenase in liver NADP* D-Fructose <———— Diet Fig. 4.10 Sorbitol pathway for synthesis of fructose. Q.22. Explain why diabetics are affected by cataract. Ans. * Fructose and sorbitol are found in lens of diabetics. * In hyperglycaemia, increased amounts of sorbitol are produced due to the increase in glucose concentration. Sorbitol cannot pass through cell membranes. Asa result, sorbitol accumulates in cells such as lens, retina, kidney and nerves, leading to cataract, neuropathy and vascular problems seen in diabetics. * Sorbitol, given intravenously, is converted to fructose. Orally given sorbitol, which forms a part of ‘sugar-free’ sweeteners, sometimes escapes absorption and gets fermented causing abdominal pain. * Aldose reductase inhibitors are being developed to retard diabetic complica~ tions such as retinopathy and neuropathy. Q.23. Explain galactose metabolism. Ans. © Galactose and glucose are produced by hydrolysis of disaccharide lactose by B galactosidase. © Lactose is an important component of milk and milk products. Also, it is synthesised only in the mammary gland of adult during lactation. =| © Galactose is also obtained from the degradation of glycoproteins and glycolip- ids. Hence, the body has ways to utilise it. * Galactose gets phosphorylated to galactose 1-phosphate by the enzyme galac- tokinase. * Galactose l-phosphate gets converted, in exchange with UDP-glucose, into UDP galactose. The enzyme that catalyses this reaction is galactose I-phosphate uridyl- transferase. * The UDP-galactose then gets converted into its C-4 epimer, UDP-glucose by UDP-hexose 4-epimerase. * This UDP-glucose is used for the formation of glycogen and UDP-glucuronate. * UDP-galactose and UDP-glucuronate are used for the synthesis of glycolipids, glycoproteins and glycosaminoglycans (GAGs). © The enzyme aldose reductase converts galactose into the alcohol galactitol (Fig. 4.11). MMMChapter 4 — Carbohydrate Metabolism Lactose B-Galactosidase NADPH + H NADPt ae \ Aldose J Galactitol Galactose reductase ATP. ‘Glycogen Galactokinase ADP Galactose 1-phosphate UDP-Glucose Galactose 1-phosphate Galactosaemia uridyltransferase Lactose ; the Glucose 1-phosphate Upplcaiationa)omiiacel ya Phosphoglucomutase +: pane a Phosphatase “pimeraee) Glucose - Glucose 6-phosphate y Liver UDP.Glucose Glycolysis, UDP.Glucuronate ¥ Proteoglycans Giycoproteins Giycosaminoglycans Fig. 4.11. Metabolism of galactose. Q24. Describe the disorders related to galactose metabolism. * Lactose intolerance occurs due to deficiency of B-galactosidase (lactase). It is found in premature infants, in conditions of mucosal cell damage. It also occurs due to surgical removal of intestine. * Nonclassical galactosaemia is caused by galactokinase deficiency. In diabetics with nonclassical galactosaemia, accumulation of galactose in lens of the eye leads to development of cataract. © Classical galactosaemia is due to the deficiency of galactose 1-phosphate uridyl- transferase. It is an autosomal recessive disorder. It results in enlargement of liver, jaundice, vomiting, mental retardation and cataract. Removal of galactose and lactose from the diet is suggested as therapy. Q.25. Describe the HMP pathway. OR “HMP pathway is called multifunctional’. Explain.Medical Biochemistry: Preparatory Manual for Undergraduates OR Why is the HMP called a ‘shunt’? Ss. * The end purpose of glucose metabolism is no Glucose can also serve as the raw biomolecules. One such mechanis: (AMP) shunt. * Synonyms of HMP shunt are phosphogluconate pathway, Warburg-Dickens Pathway and pentose phosphate pathway. It is the pathway in which glucose 64 intermediates of glycolytic pathway, Duri 5-phosphate are generated. The pentose phosphate are not used for biosyn| t always energy production. material in the synthesis of a number of m is called the Hexose Monophosphate Phosphate gets oxidised to form ing the process, NADPH and ribose pathway is called a shunt because wl hen the pentoses thetic purposes; the intermediates a Glucose 6-phosphate (6) ReDii Caliccee crohosphos NADPH4+ Htg 4 dehydrogenase (a,b) 6-Phosphogluconolactone H,0 Mat, Mn?*, Cate} 6-Phosphogluconolactone hycrolase He 6-Phosphogluconate NADPr © Phosphogluconate dehycrogenase NADPH + HY 8-Keto-6-phosphogluconate Spontaneous step co, Ribulose 5.phosphate (5C) Qecurs in [Phage OXIDATIVE Epimerase cytoplasm I NON-OXIDATIVE Isomerase Xylulose 5-phosphate (5c) Ribose 5-phosphate (50) Transketolase TPP, Mg ‘Sedohepiulose 7-phosphate (70) Phosphoglyceraldehyde (3c) Transaldolase Fructose coe Enythrose 4-phosphate m » Xyiulose S-phosphate (50) Bos Viacrensteloces TPP, Mg? Fructose 6-phosphate SPhosphoglyceraldehyde (6c) (@c) - GlycoLysis. - =— Fig. 4.12 HMP shunt pathway,Q.26. What are the salient features of HMP shunt? Ans. Chapter 4 — Carbohydrate Metabolism, * The pathway occurs in the cytoplasm. HMP shunt is active in liver, RBG, adi- pose tissue, adrenal cortex, thyroid, erythrocytes, testis and lactating mammary glands. All these are tissues which require NADPH for fatty acid biosynthesis, cholesterol synthesis or the maintenance of membrane integrity. * A notable feature of HMP shunt is that the reducing equivalent generated is NADPH and not NADH. © HMP shunt occurs in two phases (Fig. 4.12): (i) Oxidative phase * It involves generation of NADPH and ribulose phosphate. The reactions of oxidative phase are as follows: = In the first reaction, glucose 6-phosphate dehydrogenase (GPD) catalyses the irreversible oxidation of glucose 6-phosphate. This re- quires NADP* as a coenzyme. It is a regulatory enzyme which is in- hibited by a high NADPH: NADP* ratio. 6-Phosphogluconolactone is formed. ¢ In the next reaction 6-phosphogluconolactone. is rapidly hydrolysed by hydrolase to form 6-phosphogluconate. It is oxidatively decarboxylated by 6-phosphogluconolactone dehydrogenase. A pentose sugar, ribulose 5-phosphate, carbon dioxide and NADPH are the products of this reaction. (ii) Nonoxidative phase It involves conversion of ribulose 5-phosphate into ribose 5-phosphate and then to intermediates of glycolytic pathway. All the reactions are reversible. Ribulose 5-phosphate serves as a substrate for two different enzymes. ~ Isomerase that leads to the formation of ribose 5-phosphate. ~ Epimerase that helps in the formation of xylulose 5-phosphate. Ribose 5-phosphate and xylulose 5-phosphate undergo reactions cata- lysed by transketolase and transaldolase, respectively. ‘TTransketolase requires thiamine pyrophosphate (IPP). It transfers 2carbon fragment from xylulose 5-phosphate to ribose 5-phosphate releasing glyceraldehyde 3-phosphate and sedoheptulose 7-phosphate. Transaldolase, on the other hand, transfers 3-carbon fragment from sedoheptulose 5 phosphate to form erythrose 4-phosphate and glyceral- dehyde 3-phosphate. Transaldolase does not require TPP. In another reaction of carbon-fragment transfer by transketolase, a two-carbon fragment is transferred from sedoheptulose 7-phosphate to erythrose 4-phosphate. This leads to the formation of fructose 6-phosphate. The three reactions described above occur in a sequence. The enzymes involved are transketolase, transaldolase and transketolase once again. Overall reaction: 3-Glucose 6-phosphate + 6NADP — 6NADPH + 6H* + 2Fructose: 6phosphate + glyceraldehyde 3-phosphate + CO». Fructose 6-phosphate and glyceraldehyde 3-phosphate can enter glycolysis. HMP shunt is called ‘multifunctional’ because: = Although ATP is not directly consumed or generated in the cycle, HMP shunt leads to the formation of ATP as NADPH generated dur- ing this pathway is transferred to the mitochondrion where it reacts with NAD* and helps to generate ATP. = It generates reducing equivalents in the form of NADPH. - No ATP is required for the pathway once glucose 6-phosphate is formed. The pathway can continue under anaerobic conditions.“SH Medical Biochemistry: Preparatory Manual for Undergraduates = It provides ribose 5-phosphate for the synthesis of nucleotides ach nucleic acids. : nos = NADPH produced here is used in reductive syntheses (e.g. syntG7'S of fatty acids, steroids and amino acids). NADPH is also required TO reduction of oxidised glutathione (GSSG) to reduced glutathione GSH. GSH removes hydrogen peroxide (HzO2) from erythrocytes- H,0, accumulation can decrease the life span of RBCs by increasing the rate of oxidation of haemoglobin to methaemoglobin. Q.27. Describe the reasons and manifestations of GgPD deficiency. Ans. GePD deficiency. Several types of inherited deficiencies of GgPD are recognised. © It results in sensitivity to aspirin, antimalarials such as primaquine and antibi- otics such as sulphonamides, together known as the three as (aaa). * Itinduces haemolysis and anaemia. GgPD-deficient persons suffer from acute haemolytic crisis. Haemoglobin is released from the RBCs and some can be excreted in the urine. Degradation of the haemoglobin leads to the formation of Heinz bodies. © Patients who have GgPD deficiency are less likely to contract malaria. The malarial parasite (Plasmodium falciparum) infects the RBCs, where it grows on the glutathione in the cytoplasm. GPD-deficient people cannot support the growth of this parasite. Therefore, they are less prone to malaria. * GePD deficiency is seen amongst the Parsees, Kutchis and Lohana commun” ties in India. Q28. What is Wernicke—Korsakoff syndrome? Ans. Wernicke-Korsakoff syndrome. It occurs due to defective or deficient transke- tolase, resulting in ataxic stance, gait and weakness. Paralysis of eye movement and deranged mental function are also known to occur. Affinity of transketolase for TPP is reduced. The symptoms can manifest in alcoholics whose diets may be thiamine deficient. Q.29. How is HMP pathway regulated? Ans. © The first reaction catalysed by GePD is rate limiting. It is regulated by the ratio of NADP*:NADPH. * Increase in the synthesis of fatty acid and sterols causes an increase in HMP shunt. * Insulin and thyroid hormones also enhance the activity of the pathway. Q.30. What is the role of glutathione in blood? Ans. * Glutathione is y-glutamylcysteinylglycine (GSH). It is the reduced form. It is a tripeptide which helps in detoxication of HO». * NADPH is used by glutathione reductase to generate reduced glutathione (Fig. 4.13). * Reduced form of glutathione helps in: * Oxidation of H;0>. * Prevention of auto-oxidation of lipids. * Amino acid transport in the y-glutamyl cycle. * The glutathione defence does not work adequately in GgPD deficiency, infections, oxidative stress caused by certain drugs and when fava beans are ingested. Fava beans are found in the Mediterranean regions.Chapter 4 — Carbohydrate Metabolism 6-Phospho- ; @s-sG ‘gluconate NADPH + H (oxidized) 2H,0 + Of i Glutathione [GERD deficiency] Glutathione reductase (Pecoxniese, Ger NADPt 2GSH 2H,0,4¢— [Oxi (reduced) So | [Haemolysis] [Heinz bodies] <——Hb aggregates. Fig. 4.13 Manifestation of glucose 6-phosphate dehydrogenase deficiency. © The oxidised form is written as GSSG. © Glutathione precursors such as N-acetyl cysteine are used in therapeutics as antioxidants. Q31. Write a note on the uses and regulation of NADPH. Ans. Following are the uses and regulation of NADPH: Uses of NADPH * It causes reduction of glutathione. * Detoxication using cytochrome P-450 system. ¢ NADPH is required for the microsomal-cytochrome P-450 monooxygenase system. © generation of nitric oxide and of reactive oxygen species by phagocytes ° scavenging of reactive oxygen species that form as byproducts of oxygen transport and of the respiratory chain * NADPH is required for synthesis of fatty acids, steroids, neurotransmitters and nucleotides. Regulation of NADPH * Cellular concentration of NADP* is the major factor regulating the pathway. The availability of NADP* regulates the rate-limiting reaction catalysed by the GePD. * Thus, the path taken is largely determined by the needs of the cell for NADPH or sugar intermediate. NADP+H* NADP* RH Hydroxylated substrate (ROH) O2 Cytochrome P-450 Q.32. What are the salient features of uronic acid pathway? Ans © Uronic acid pathway is another synthetic pathway involving glucose. * Itis also called glucuronic acid cycle. * Ivis an alternative pathway for the oxidation of glucose.56 Medical Biochemistry: Preparatory Manual for Undergraduates * It provides p-glucuronic acid which is used for detoxication and synthesis Of mucopolysaccharides. * The pathway is a source of UDP-glucose used in glycogen metabolism. The pathway operates in adipose tissue besides others. Reactions of uronic acid pathway are shown in Fig. 4.14. Q.33. State the salient features of the uronic acid pathway. ‘ Ans. The uronic acid pathway does not appear to be very essential for humans, as individuals with blocks in the pathway do not suffer major ill effects. The important aspects to be remembered are as follows: Phosphoglucomutase UDP-Glucose pyrophosphorylase Glucose ¢ P< Glucose 1-phosphate< =< Uridine phosphate loose (UOP6) UTP BP, t anno | rH,0 UpP-Glucose dehydrogenase 2NADH + 2Ht UDP-Glucuronic acid ; D-Glaeuronie acid] —> GAGs ot + / NADP 3 L-Glucuronate . reductase :Kelo-L-gulonate decarboxylase et-OH acid dehydrogenase Y Lxylulose 3-Keto-L-gulonio L-Gulonie acid acid co, NAD? NADH + Ht NADPH + HY Xylulose 20, reductase Aldonolactonase NADP* (Pantosura Gulonolactone Xylitol NaD* 02 Gulonolactone oxidase Xylitol dehydrogenase Block in primates and guinea pigs = NADH + HY 1 D-Xylulose [2:Keto-L-gulonolactone] ATP Xylulose Winase \ L-Ascotbie acid D-Xylulose 5-phosphate MP shunt Fig. 4.14 Uronic acid pathway.Chapter 4 — Carbohydrate Metabolism 57 | * Humans and guinea pigs lack the enzyme that converts t-gulonolactone to L-ascorbic acid. Hence, vitamin C needs to be consumed by humans. * Deficiency of t-xylitol dehydrogenase and xylulose reductase causes pentosuria due to accumulation of xylitol, a pentose sugar. © Drugs such as barbital and aminopyrine increase uronic acid pathway. © glucuronic acid formed is used for detoxication and synthesis of mucopoly- saccharides. Q.34. Explain the oxidation of pyruvate to acetyl-CoA. OR What is the central role played by acetyl CoA? Ans. The end product of aerobic glycolysis is pyruvate. However, pyruvate is not the end product of carbohydrate metabolism. There is still precious energy inside the pyruvate molecule, and the body will not let that energy go waste. In order to use that energy, the pyruvate molecule is then routed through a pathway known as the tricarboxylic acid cycle. © For this to be done, pyruvate is first converted to acetyl CoA. ® Acetyl CoA then enters TCA cycle. This reaction is important because it pro- vides the link between glycolysis and TCA cycle. ® It occurs in mitochondrial matrix. © The reaction can be represented as: Pyruvate + NAD* + CoASH — Acetyl CoA + NADH + H* + CO. * Pyruvate is oxidatively decarboxylated with the help of a pyruvate dehydroge- nase (PDH) complex which is a multienzyme complex. This complex is made up of three enzymes and requires five coenzymes. The enzymes are as follows: E,: pyruvate decarboxylase Es: dihydrolipoyl transacetylase Es: dihydrolipoyl dehydrogenase * Coenzymes required are TPP, lipoic acid, FAD, NAD* and CoA. © The reactions of TCA cycle are irreversible, rate-limiting and regulatory. © Regulation of reaction: * Increased by insulin * Decreased by high ratios of ATP: ADP and NADH: NAD*, presence of acetyl CoA and cAMP and conditions of starvation and diabetes mellitus. Q.35. Describe the manifestations of pyruvate dehydrogenase complex deficiency. ‘Ans, Defect in the PDH complex is fatal in some cases. Symptoms include lactic acido- sis and neurologic disorders. Treatment involves large doses of thiamine, lipoic acid and ketogenic diet. It is hence, referred to as thiamine responsive PDH deficiency. Q.36. Give the details of the tricarboxylic acid cycle? State its salient features. OR Write a brief note on reactions involved in the TCA cycle. Ans. © Tricarboxylic acid (TCA) cycle is a series of reactions through which final oxidation of acetyl coenzyme A (acetyl CoA) to CO2 occurs. Thus, TCA cycle requires acetyl CoA as a substrate. © The acetyl group of the acetyl CoA is derived from the other pathways of metabolism such as oxidation of fatty acids, ketone bodies, proteins and most importantly, glycolysis.Medical Biochemistry: Preparatory Manval for Undergraduates Salient features of TCA cycle ¢ Symonyms are citric acid cycle, Krebs cycle (Fig. 4.15). * Site of occurrence is mitochondrial matrix. * TOA: cycle is called ‘amphibolic’ because both catabolic and anabolic Processes are involved in this pathway. * Catabolic process: The cycle helps in the degradation of acetyl residues, which are derived from carbohydrates, fats, proteins, etc. * Anabolic process: The intermediates of TCA cycle are used as precursors in the biosynthesis of many compounds. * TGA cycle is a common pathway for the oxidation of carbohydrates, fatty acids and amino acids to carbon dioxide and water. It provides electrons which are transferred to electron transport chain where they help in the formation of ATP. * Oxaloacetate is regenerated through TCA cycle, Therefore, it is said to play a catalytic role. Pyruvate NAD* Pyruvate dehydrogenase complex NADH + Ht AcetylCoA —CoASH H,0 Citrate Oxaldacetate Synthase Citrate NADH + Ht, Fe Malate Aconitase NAD* dehydrogenase TEROrSScSIaTa| ne) LMalate {Cis-Aconitate) Fu Fe% smarase Aconitase\ \~» 4,0 H,0 Fumarate Isocitrate FADH, CO, Succinate NAD* Isocitrate ehydrogenase Malonate dehydrogenase FAD ee NADH + Ht ‘Succinate ocKetoglutarate Succinate thiokinase e-Ketoglutarate C0, dehydrogenase ‘ complex NAD nines NADH + Ht Succinyl CoA COASH Fig. 4.15 TCA cycle.Chapter 4 — Carbohydrate Metabolism — © Vitamins play an important role in the TCA cycle. * Riboflavin is required for reactions catalysed by a-ketoglutarate and succinic dehydrogenases. ° Niacin is required for isocitrate-, a-ketoglutarate- and malate dehydrogenases. * Thiamine is required for a-ketoglutarate- and pyruvate dehydrogenases. * Pantothenic acid is required for synthesis of succinyl CoA. © Carbon dioxide which is lost in the reactions catalysed by pyruvate-, a-ketoglutarate and isocitrate dehydrogenases is transported to the lungs for exhalation. © The pyruvate dehydrogenase and a-ketoglutarate dehydrogenase complexes are similar. * Succinate thiokinase is involved in substrate level phosphorylation. GDP is converted to GTP. GTP is used to produce ATP GTP + ADP —— GDP + ATP © Overall reaction of TCA cycle is Acetyl CoA + 8NAD* + FAD + GDP + P; + 2H;0 —> 2CO, + CoASH + 3NADH + 3H* + FADH, + GTP © All the enzymes of the TCA occur in the mitochondrial matrix except the suc- cinate dehydrogenase, which is found in the inner mitochondrial membrane. In the first reaction of TCA cycle, acetyl CoA combines with oxaloacetate to form citrate with the help of citrate synthase. Citrate inhibits this reaction. Citrate undergoes isomerisation with the help of aconitase to produce isocitrate. Isocitrate is oxidised to a-ketoglutarate in an oxidative decarboxylation reac- tion forming carbon dioxide. Isocitrate dehydrogenase complex is involved in this reaction. It is a key regulatory enzyme. It is activated by ADP and inhibited by NADH. a-Ketoglutarate is converted to succinyl CoA in the second oxidative decarbox- ylation reaction. Carbon dioxide is released and NADH* and H* are produced in this reaction. Enzyme required is a-ketoglutarate dehydrogenase complex, which is similar to pyruvate dehydrogenase complex. Succinyl CoA is cleaved to succinate with the help of succinate thiokinase. This is a reaction which involves substrate level phosphorylation. Consequently, GDP is converted to GTP. This GTP is used for protein, DNA synthesis and RNA synthesis. The GTP is also used to produce ATP. Succinate is now oxidised to fumarate. This reaction is catalysed by succinate dehydrogenase. FAD is the coenzyme required, this reaction yields 2 ATPs. Hydration of fumarate generating malate occurs with the help of fumarase. Malate is oxidised with the help of malate dehydrogenase. NAD* is required as a coenzyme. Three ATPs are formed as the electrons pass through the electron transport chain. Oxaloacetate is regenerated in this reaction. ° Q.37. Explain the amphibolic role of TCA cycle. Ans. The word ‘amphi’ means both. TCA cycle, and both the catabolic and anabolic reactions are involved. Therefore, it is called ‘amphibolic’ (Fig. 4.16). Anabolic role (a) Amino acid metabolism Anumber of glycogenic amino acids enter the TCA cycle via transamina- tion reactions Glutamate > -Ketoglutarate Aspartate > Oxaloacetate Alanine > PyruvateAcetyl CoA, it Fatty acid |<—— Oxalodcetate Citrate | thesis TCA j<—Malate cycle [Amino acial egress ml riba Succinyl CoA [Neurotransmitter| Haem synthesis} Fig. 4.16 Anabolic role of TCA cycle. (b) Lipid metabolism * Acetyl CoA and citrate are required for the synthesis of steroids and fatty acids. (©) Porphyrin synthesis * Succinyl CoA and glycine are used for the synthesis of porphyrin. It is thus, involved in the synthesis of haemoglobin, cytochromes and haemo- proteins. (a) Nitrogen metabolism © The aspartate produced by the transamination of oxaloacetate is used for the synthesis of argininosuccinate and purines. The end product of these processes is fumarate, which then enters the cycle. Catabolic role The oxidation of acetyl CoA produces energy, COs and water. Q38. Explain the energetics taking place in TGA cycle. OR Tabulate the formation of ATP during one turn of TCA cycle. Ans. * One tum of the TCA cycle, starting with acetyl CoA, produces 12 ATPs (Table 4.4). * When the starting molecule is pyruvate, the oxidative decarboxylation of pyru- vate yields 3 ATPs. © Therefore, 15 ATPs are produced when the starting compound is pyruvate. © The reaction involving succinate thiokinase, a classical example of substrate level phosphorylation (SLP), generates a high-energy phosphate bond of GTP which is later converted into ATP. © Since two molecules of pyruvate enter the TGA cycle when glucose is metabolised, the number of ATPs is doubled. Therefore, 30 ATP molecules are produced when pyruvate enters the TCA cycle. The formation of ATP during one tum of TCA cycle is as shown in Table 4.4. Q.39. How is TCA cycle regulated? Ans. ¢ The regulatory enzymes of the TCA cycle include citrate synthase, isocitrate dehydrogenase and -ketoglutarate dehydrogenase complex.Chapter 4 — Carbohydrate Metabolism Reaction catalysed by ATP production Pyruvate dehydrogenase complex NADH via ETC Isocitrate dehydrogenase NADH via ETC. eKetoglutarate dehydrogenase complex _ NADH via ETC Succinate thiokinase SLP Succinate dehydrogenase FADH, via ETC Malate dehydrogenase NADH via ETC 15 ATP x 2 ETC, electron transport chain. ° High ratios of ATP:ADP, acetyl CoA:CoA and NADH:NAD* inhibit the cycle. Citrate, fluoroacetate and arsenite are inhibitors of specific reactions. © Insulin stimulates the pyruvate dehydrogenase complex. * Oxaloacetate has a catalytic role to play in stimulating the entry of acetyl CoA into the TCA cycle. Thus, the availability of oxaloacetate appears to regulate the activity of the cycle. Q.40. Briefly elaborate on deficiencies of the TCA cycle. Ans. * Deficiencies of enzymes of TGA cycle are rare, However, mitochondrial en- cephalopathy occurs due to fumarase deficiency. It is an autosomal recessive disorder and elevated levels of succinate, a-ketoglutarate, citrate and malate occur, It is characterised by a mitochondrial myopathy affecting both the skeletal muscle and the brain. The symptoms include severe neurological impairment, encephalopathy and dystonia. Urine contains abnormal amounts of fumarate. Q.41. What is gluconeogenesis? Add a note on substrates required for gluconeogenesis. Ans. © Definition: It is the process by which synthesis of carbohydrates from noncarbo- hydrate precursors occurs. Precursors. Lactate, pyruvate, glycerol and glucogenic amino acids. Site, It occurs mainly in liver. The liver makes about 85%-95% of the glucose. The kidney contributes about 50% of glucose during starvation. The only other tissues involved in gluconeogenesis is those of small intestine. © Compartmentalisation: Some of the reactions of gluconeogenesis occur in the mitochondria and the other in the cytoplasm. Pathway. Starting with pyruvate, most of the steps of gluconeogenesis represent a reversal of glycolysis except at four points (Fig. 4.20). The four enzymes unique to gluconeogenesis when compared with glycolysis are pyruvate carboxylase (mitochondrial), PEP carboxykinase (cytoplasmic), fructose 1,6-phosphatase (cytoplasmic), glucose 6-phosphatase (cytoplasmic). © Substrates for gluconeogenesis: (i) LactateMedical Biochemistry: Preparatory Manual for Undergraduates BLOOD LIVER Glycogen muSCLE| Glucose Glucose - Pyruvate Pyrlvate & NADH + Ht NADH+H*| 2 ‘ LOH LDH ‘ 8 NAD* NAD* s & & Lactate < Lactate Alanine < Alanine Fig. 4.17 Cori's cycle and glucose-alanine cycle. Itis transported to the liver by the Cori’s cycle and is converted to pyru- vate (Fig. 4.17). Lactate is formed from glucose by glycolysis in the muscle cells. Lactate is released into the blood and taken up by the liver. Lactate is converted to glucose via gluconeogenesis in the liver. © Glucose is released back into the circulation. (ii) Alanine © Pyruvate is converted to alanine by transamination reactions in the muscle cells via the glucose-alanine cycle (Fig. 4.17). ° The alanine is transported from the muscle to the liver, where it is converted back to pyruvate. © Pyruvate is then converted to glucose via gluconeogenesis in the liver. (iii) Glycerol © It is formed in the adipose tissue by lipolysis of triacylglycerols and is released into the blood and taken up by the liver. © In the liver, glycerol is converted to glycerol 3-phosphate, which is finally converted into glyceraldehyde 3-phosphate. © Glyceraldehyde 3-phosphate is then converted to glucose via gluconeo- genesis. (iv) Other amino acids (Fig. 4.18) * Anumber of amino acids are conyerted to intermediates of TCA cycle which then enter the gluconeogenesis pathway. (v) Propionate (Fig. 4.19) * Propionic acid, a product of odd-chain fatty acid oxidation, is converted to succinyl CoA, which then enters the TGA cycle. Q.42. What is the significance of gluconeogenesis? Ans. The significance of gluconeogenesis is as follows: * Blood sugar level is maintained via gluconeogenesis. * d-glucose is absolutely necessary for the tissues such as brain, erythrocytes, kidney and eyes. * The kidneys help in the excretion of increased number of protons during metabolic acidosis.Chapter 4 — Carbohydrate Metabolism ee Pyruvate < Alanine « —— Tryptophan. on — ‘Oxaloacetate, Malate aa S o-Ketoglutarate < ——— Proline Aspartate ——————> Fumarate Glutamate oz Histidine mpi ‘Succinate <— Valine ‘Tyrosine < Phenylalanine Cysteine Aspartate Threonine Methionine Isoleucine Fig, 4.18 Entry of amino acids into TCA cycle. CO, acyl CoA Propionyl CoA synthase carboxylase. EEC ea came Propionyl CoA D-Methylmalonyl CoA Biotin ATP PP, ATP AMP-PP; ‘Methylmalonyl CoA racemase ‘Methylmalonyl CoA mutase ~ = Vit. Biz Fig. 419 Point of entry of propionate. Suecinyl CoA D-Methylmalonyl CoA * Glucose 6-phosphatase is absent in muscle. Therefore, during exercise and starvation, the large amounts of lactate produced by glycolysis and glycerol generated by lipolysis of triacylglycerols are used up by gluconeogenesis. * Some amount of protein (in the form of carbon skeleton of glucogenic amino acids) is also converted to glucose. Thus, a proper interplay between the pro- tein, carbohydrate and lipid metabolic pathways occurs. Q43. Write a short note on the reactions of gluconeogenesis. Ans. Only those reactions that are not common to glycolysis are given below. These reactions are reactions (1)—(4) in Fig. 4.20. Reaction (1): Carboxylation of pyruvate © Pyruvate is converted to oxaloacetate with the help of carbon dioxide, ATP and pyruvate carboxylase. Biotin, Mg?* and Mn®* are required by the enzyme for its activity. There is also an absolute requirement for acetyl CoA.Medical Biochemistry: Preparatory Manual for Undergraduates Glucose 0 lucose 6-phosphatase fexokinase (or) = Glucose 6- Glucokinase a ® Et Fructose 6+ Oran Fructose 1/6-bi i” Phosphofructokinase @ Citrate (o/ \ Fructose 1,6-bisphosphate © AMP; F2,6-BP. Fiat t ADP (2) Glyceraldehyde 3-phosphate <> DHAP NADH + H*4>) Glycerol 3-phosphate Nene dehydrogenase | Glycerol 3-phosphate (2) 1,3-Bisphosphoglycerate ADP. Glycerol kinase ATP (2) 34 a (2) a Phosphoenol (2) Phosphoenol pyruvate pyruvate [carboxykinase ‘ADP GDP +CO, ATP Pyruvate ——> ce { Mitochondrion Pyruvate as +CO, ruvate cat lase Oxaloacetate Sie +p, (a rboxyle Mn? AS @Acetyl CoA NAD* Biotin ‘NAD* ADP. Malate 2 f a aY. pn eae: / Succinyl CoA<—C Propionate > Entry of amino acids Fig. 4.20 Pathway of gluconeogenesis.Chapter 4 — Carbohydrate Metabolisr ° This reaction occurs in the mitochondrial matrix. * Oxaloacetate produced in the mitochondrion cannot cross the membrane. It is first reduced to malate, which then moves across the mitochondrial membrane into the cytosol. Malate is reoxidised to oxaloacetate in the cytosol. Reaction (2): Conversion of oxaloacetate to phosphoenolpyruvate (PEP) © The enzyme required is phosphoenolpyruvate carboxykinase (PEPCK). It requires Mn** and GTP. Along with PEP, carbon dioxide and GDP are formed. © This reaction occurs in cytosol. * Reversal of the reactions of glycolysis now occurs until F1,6BP is formed. Reaction (3): Dephosphorylation of F1,6BP It produces fructose 6-phosphate and is catalysed by fructose 1,6-bisphosphatase which is the major regulatory enzyme in gluconeogenesis. This enzyme is activated by citrate and inhibited by adenosyl monophosphate (AMP) and F2,6BP. Fructose 6-phosphate is converted to glucose 6-phosphate as in glycolysis. Reaction (4): Dephosphorylation of glucose 6-phosphate © This reaction gives free glucose and inorganic phosphate. ® It is catalysed by glucose 6-phosphatase which is present only in the liver, kidney and epithelial cells of the small intestine. Q.44. How is gluconeogenesis regulated? Ans, Regulation of gluconeogenesis © Gluconeogenesis is dependent on concentration of the precursors. * Acetyl CoA is a positive regulator of pyruvate carboxylase. It activates the enzyme. * Fructose 2,6-bisphosphatase is inhibited by AMP. * Hormones such as glucagon, epinephrine and glucocorticoids stimulate the process, * Insulin suppresses gluconeogenesis. © Thus, gluconeogenesis is stimulated during fasting, prolonged exercise, by a high protein diet and times of stress. Q45. Explain the energetic involved in gluconeogenesis. Ans. Two molecules of pyruvate are converted to one molecule of glucose. To help this conversion, six molecules of ATP are required. Q.46. What is multiple carboxylase deficiency? Ans. Multiple carboxylase deficiency occurs due to a genetic defect in the enzyme holo- carboxylase synthetase which is responsible for binding biotin to pyruvate carboxyl- ase and to other carboxylase enzymes, thus affecting their activity. It is reflected in. developmental retardation, ketoacidosis, hair loss and erythematous rash. Q.47. What is von Gierke disease? Explain the biochemical manifestations. Ans. It is glycogen storage disease type I. Itis caused due to the defect of glucose 6-phosphatase. Hepatomegaly, lactic acidosis, hyperuricaemia, hyperlipidaemia and hypoglycaemia are the manifestations. Glucose 6-phosphate produced by gluconeogenesis accumulates in the liver, activating glycogen synthesis. This leads to large deposits of glycogen in the liver causing hepatomegaly and also hypo- ghcaemia. Accumulation of glucose 6-phosphate due to the deficiency of glucose 6-phosphatase leads to increased glycolysis causing increase in pyruvate and lactate. This causes lactic acidosis. Since gluconeogenesis is blocked, the body relies on lipid metabolism for energy. Hyperlipidaemia results. The large amounts of glucose 6-phosphate are shunted into the HMP pathway, which leads to excessMedical Biochemistry: Preparatory Manual for Undergraduates pentose sugars production. PRPP synthesis increases, causing degradation of purines and eleyated uric acid levels, causing hyperuricaemia. Q48, What are the ‘anaplerotic reactions’ or ‘filling up’ reactions in gluco- neogenesis? Ans. © The intermediates of the TGA, such as oxaloacetate, a-ketoglutarate, citrate and succinyl CoA are used for the biosynthesis of other molecules. © Their utilisation leads to the removal of these molecules, resulting in the slowdown of the cycle. Therefore, these intermediates need to be replaced or replenished. * It is achieved by the following replacement or anaplerotic reactions: Biotin 1, Pyruvate + CO) ++ ATP ————> Oxaloacetate + ADP + Pi Mg? Catalysed by pyruvate carboxylase, this reaction occurs in mitochondria. It is a part of gluconeogenesis. Malic enzyme 9. Pyruvate + COs + NADPH + Ht <-> t-malate + NADP+ Malic enzyme occurs in the cytoplasm and converts pyruvate into malate. Malate then enters into the mitochondria as a substrate for the TCA cycle. 8. Oxaloacetate and a-ketoglutarate are obtained from aspartate and gluta- mate by transamination reactions. Me 4, Phosphoenolpyruyate + GDP + CO. <> Oxaloacetate + GIP The enzyme involved is phosphoenolpyruvate carboxykinase and is a part of the gluconeogenic reactions. Oxaloacetate is replenished by this reaction. 5, Glutamate is reversibly converted to a-ketoglutarate by glutamate dehydro- genase. 6, Valine, isoleucine and propionyl CoA all get converted into succinyl GoA and enter TGA cycle. Q49. Explain glycogen metabolism. State its salient features. Ans. © Glycogen is polymer of glucose molecules. It is the storage form of glucose. It acts as an immediate source of blood glucose for use as metabolic fluid. * Glycogen exists as granules in the cytosol. © It mainly occurs in muscle and liver. * In muscle it acts as a source of energy and in liver it helps in shortterm maintenance of blood glucose homeostasis. © The metabolism of glycogen involves: © Synthesis of glycogen (glycogenesis) © Utilisation of glycogen (glycogenolysis) Salient features of glycogenesis (Fig. 4.21) © Glycogen is synthesised in cytoplasm of muscle and liver cells. © The glycogen stores in the liver are adequate for 12 hours or less without gluconeogenesis. ® Glycogen molecule is highly branched. Glucosyl units are linked by « (14) bonds with a (1-96) branches after every 8-10 residues.Chapter 4 — Carbohydrate Metabolism Phosphoglucomutage Glucose 6-phosphate <= Glucose 1-phosphate + UTP Mg UDP-Glucose pyrophosphorylase UDP-Glucose + PP, © cAMP @00- ® Glucose, insulin ae Giycogenin (primer) @000- + UDP ° Glucosy-4,6 transferase e@eoe0000 «<——_—___ eeeee00- Glycogen (1 —4) Glycosyl units added Fig. 4.21 Glycogenesis. © UTP is required for glycogen synthesis. © Advantages of branching in glycogen include: ® Increased solubility of glycogen. * Increase in number of nonreducing ends to which new residues can be added or removed. Q.50. What are the reactions involved in glycogenesis? Ans. In the first reaction, glucose 6-phosphate is converted to glucose 1-phosphate with the help of phosphoglucomutase. Glucose 1-phosphate reacts with UTP to form UDP-glucose. This reaction is catalysed by UDP glucosepyrophosphorylase. Inorganic pyrophosphate (PP;) is released in this reaction. Initiation of glycogen synthesis requires a primer. Glycogen primer is made up of 4-5 glucose units. The primer is attached to a protein, glycogenin. It acts as an acceptor of glucose units. The reactions of elongation of the glycogen chain is catalysed by glycogen synthase. It helps in the formation of a (1—>4) linkages. This is a rate-limiting step. Glycogen synth UDP-Glucose + Glycogen (n) > UDP + Glycogen (n+ 1) Formation of branches of glucogen occurs with the help of glucosyl-4,6 transferase. Cascade regulation of glycogen synthesis is represented in Fig. 4.22. Glycogen synthase is present in two forms: (i) Glycogen synthase D, which is inactive and is dependent on concentration of glucose 6-phosphate. (ii) Glycogen synthase 1, which is active and is independent of glucose 6-phosphate. The interconversion of D and I forms is catalysed by protein kinase which is dependent on the presence of hormones, epinephrine and glucagon. © Asa result of the inhibition of glycogen synthase, glycogenesis is inhibited.Medical Biochemistry: Preparatory Manual for Undergraduates Epinephrine Adenylate cyclase —____-» Adenylate cyclase (active) (inactive) Glucagon. ‘ATP ——*—> cAMP Protein kinase ———+ Protein kinase (active) (inactive) ATP ADP Glycogen synthase |» Glycogen synthase D (active) (inactive) Insulin UDP-Glucose —* + Glycogen (synthesis inhibited) Fig. 4.22 Cascade for control of glycogenesis. Q.51. What are the disorders associated with glycogenesis? Ans. The disorders associated with glycogenesis are as follows: © Lewis disease occurring due to deficiency of glycogen synthase. ‘© Andersen disease occurring due to deficiency of glucosyl-4,6 transferase. © Glycogen storage diseases occur due to deficiencies in certain enzymes of glyco- gen metabolism, These deficiencies lead to accumulation of glycogen and/or inability to use glycogen as a fuel. Infants born with these diseases suffer cirrhosis and liver failure. Most infants die by two years of age. Q52, What are the salient features of glycogenolysis? Ans. The salient features of glycogenolysis are as follows: * Itoccurs in cytoplasm of muscle and liver, * Glycogenolysis involves degradation of glycogen by glycogen phosphorylase which yields glucose -phosphate and a glycogen chain that is smaller by one glucose unit. © Glycogen phosphorylase cannot act on the four glucosyl residues closest to the a (1-96) branch point (Fig. 4.23). _— @ +Glucose Glycogen 40-0-Glucano- phosphorylase transferase -1,6-Glucosidase SS Oe (debranching enzyme) Fig. 4.23 Glycogenolysis.Chapter 4 — Carbohydrate Metabolism (Muscle and liver) Epinephrine/Glucagon Adenylate cyclase Adenylate cyclase (active) (inactive) ATP ——> cAMP Protein kinase —~ » Protein kinase (inactive) (active) Phosphorylase kinase ———» Phosphorylase kinase (inactive) (active) Phosphorylase kinase b ____» Phosphorylase kinase a (inactive) (active) Glycogen (n) > Glucose 1-phosphate + Glycogen (n—1) Fig. 4.24 Amplification of cascade for control of glycogenolysis. © A debranching enzyme system removes the glucosyl residues near each branch point. Ithas two catalytic activities: itacts as a 4:4 glycosyltransferase and 1:6 glucosidase. ° The 4:4 transferase removes a fragment containing three glucose units and then attaches them to the end of long chain by « (1—>4) bond. Then the 1,6glucosidase of the debranching enzyme system acts to release free glucose. Thus, free glucose and about 7-9 glucose I-phosphate molecules are released. Regulation by cAMP-mediated cascade involving phosphorylase is represented in (Fig. 4.24). Glycogen phosphorylase exists in two forms: * An active phosphorylase a, which is the phosphorylated form. * An inactive phosphorylase b, which is the dephosphorylated form. The interconversion of these two forms is dependent on phosphorylase kinase, which is in turn regulated by hormones, epinephrine, glucagon and insulin. Thus, an elevated insulin level results in increased glycogen synthesis, whereas an elevated glucagon (or epinephrine) level causes increased glycogenolysis. cAMP levels alter in response to the hormones. In the fed state, glycogen synthase is activated by glucose 6 phosphate while glycogen phosphorylase is inhibited. Q.53. What are the disorders associated with glycogenolysis? Ans. The disorders associated with glycogenolysis are as follows: * Type II glycogen storage disease (Pompe disease) caused by a defect in « (14) glucosidase. Symptoms of this disease are psychomotor retardation, an en- larged heart and eventual failure of heart and lungs. * Type III glycogen storage disease (Cori disease ot Forbes disease) caused by the defect of debranching enzyme system. This disease also causes heart and lung problems, stunted growth, enlarged liver, hypoglycaemia and acidosis.: Preparatory Manual for Undergraduates Q54. How is glycogen metabolism regulated? OR Explain the differences in glycogen metabolism of muscle and liver. Ans. The regulation of glycogen metabolism in different tissues depends on the function of glycogen in them. * In muscle, glycogen acts as a reseryoir-which can be used for generation of ATP by oxidation of glucose. In this case: ¢ Epinephrine promotes glycogenolysis and inhibits glycogenesis. * Insulin increases glycogenesis and decreases glycogenolysis. © cAMP and Ga®* released also regulate glycogen metabolism. * In liver, glycogen serves to support supply of blood glucose during fasting. In this case: * Glucagon increases glycogenolysis and decreases glycogenesis. * Insulin increases glycogenolysis by increasing the concentration of glycogen synthase. * High insulin:glucagon ratio provides for storage of glycogen after a meal. Increased levels of blood glucose increase glycogenolysis. * Low levels of cAMP increase glycogenolysis and decrease glycogenesis. * The modulation of various pathways is described below. Activator Inhibitor Glycogenesis Insulin, glucose 6-phosphate Glucagon Glycogenolysis Glucagon, epinephrine, glucocorticoids Insulin | Gluconeogenesis Glucagon, glucocorticoids, acetyl CoA Insulin, ADP J Q.55. Write a note on the regulation of blood sugar OR Explain the role played by the different organs in the maintenance of normal blood sugar. OR Write a note on the hormonal regulation of blood sugar. Ans. Obviously, glucose has an extremely important role to perform in the body. However, if the levels of the free glucose in the blood are higher than a certain level, especially for longer periods of time, it can have dire consequences. Thus, the level of glucose in the blood must be maintained within a specific spectrum, of concentration. We shall now see how this level is maintained through the regulation of blood sugar (Fig. 4.25). (a) Role of organs in regulation of blood sugar. ©) Liverinfluences the blood sugar through the following biochemical reactions. ® Glucose is freed by the action of glucose 6-phosphatase. ° Cori’s cycle. ® Glucose-alanine cycle * Gluconeogenesis * Skeletal muscle. The uptake of glucose is influenced by insulin. Since glucose 6-phosphatase is absent here, free glucose cannot be released.Chapter 4 — Carbohydrate Metabolism Intestine and liver Muscle Absorption of molecules Glycogenesis Interconversion Brain Oxidation to supply a so Blood glucose |______» Synthesis of lactose, Glycogenolysis, (60-100 mg) glycolipids, mucopolysaccharides, epinephrine, glucagon nucleic acids phosphorylase, glucose 6-phosphatase Synthesis of non-amino acids Liver Adipose tissue Gluconeogenesis Cori’s cycle Conversion to fat for storage Glucose—alanine cycle Excretion in urine (abnormal) Fig. 4.25 Regulation of blood glucose. © Adipose tissue Hydrolyses of triacylglycerols to release glycerol takes place. Glycerol is taken to liver for gluconeogenesis as it cannot be phosphorylated due to absence of glycerol kinase in the adipose tissue. (b) Roles of hormones in regulation of blood sugar. ° Insulin is secreted by the B-cells of pancreas. It is released by amino acids, free fatty acids, ketone bodies, glucagon, secretin and tolbutamide. Epinephrine and norepinephrine block the release of insulin. Insulin transporters increase glycolysis by stimulating phosphofructokinase. Glu- coneogenesis, glycogenolysis, ketogenesis are decreased whereas lipogen- esis and protein synthesis are increased by insulin. © Anterior pituitary hormones: Growth hormone (GH) is stimulated by hypogly- caemia and causes decreased uptake of glucose while adrenocorticotro- phin (ACTH) mobilises free fatty acids from adipose tissue and produces hyperglycaemia. © Adrenal cortex hormones: Glucocorticoids such as cortisol act antagonistic to insulin, Therefore: © Gluconeogenesis increases. © Utilisation of glucose occurs in extrahepatic tissue. ° Increased formation of glucose in liver by stimulating glucose 6-phos- phatase and fructose 1,6-biphosphatase occurs.as " Medical Biochemistry: Preparatory Manual for Undergraduates © Epinephrine: * In muscle, glycogen breakdown is aided by phosphorylase which, in turn, is activated by epinephrine. The glucose formed by degradation of glyco- gen is converted into lactic acid via the Cori’s cycle. There is decreased release of insulin. * In the liver, glucose gets utilised. Glucagon is secreted by the accells of pancreas. It is stimulated by hypoglycae- mia. It causes glycogenolysis in the liver: Its action is similar to that of epineph- rine. It stimulates glucose 6-phosphatase and thus enhances gluconeogenesis. © Thyroid hormones stimulate glycogenolysis. * Sex hormones such as estrogens are responsible for decreased levels of insulin and thereby a decrease in the blood sugar levels. Q'56. Write a short note on the Glucose Tolerance Test (GTT). Ans. The GTT is carried out only under certain indications as follows: ‘© Indications © To establish the presence of diabetes mellitus in patients who show normal blood sugar levels in either fasting or postlunch conditions. + During pregnancy. * In the evaluation of health of an obese person. * To investigate glycosuria. * The test is done in the following way. * The individual is kept fasting for 10-12 hours. * Samples of venous blood and urine are collected. * 50-70 g of glucose are given orally with lemon juice. The blood sample is drawn every 30 minutes for 2-8 hours. * Interpretation of GTT (Fig, 4.26) is done as follows: * In nondiabetic individuals, all urine specimens show a negative reaction to Benedict’s qualitative reagent. URINE BLOOD Severe >2% glucose 320-350 mg Mild 1-2% glucose 190-200 mg 400: Pre-diabetes — —ve urine 150-160 mg Normal =ve urine 100 mg E200 co ‘Severe 160: He tee Mid ® 75 Pre-diabetes Normal ° 12 1 Awe 2 Time in hours Fig. 4.26 Glucose tolerance test. 21 3Chapter 4 — Carbohydrate Metabolism © The peak glucose levels are normally achieved within 30-60 minutes and may increase to 90 mg/dL. © The mild diabetics show a peak within 30-60 minutes, but a severe diabetic may reach the peak level later than this time period. © Insulin secretion from the pancreas is stimulated by glucose absorption. As a result, the blood glucose levels drop below the fasting level after 30-60 minutes and return to normal after 90-120 minutes. Q.57. Write a note on the biochemical aspects of diabetes mellitus. Ans. * Diabetes mellitus is a chronic, metabolic disorder that manifests as hypergly- caemia and glycosuria and is characterised by a relative or absolute lack of insulin, resulting in altered carbohydrate, fat and protein metabolism. Fig. 4.27 represents the biochemical manifestations. © Symptoms are polyuria, polyphagia, polydipsia, sudden loss or weight, asthenia, weak- ness, nonhealing wounds, ketoacidosis causing acetone breath, and infections. * Causes © Congenital, i.e. hereditary * Acquired © Surgical, i.e. due to pancreatectomy, splenectomy Medical, i.e. destruction of B-cells by alloxan Decreased insulin secretion by B-cells (IDDM) Sensitivity of target cells to insulin as in obesity (NIDDM) Autoimmunity against B-cells of pancreas Insulin ¥ ¥ and glucagon * + Glucose uptake 4 Proteolysis 4 Lipolysis by liver Hyperglycaemia + Plasma 4 Plasma FFA and Glucosuria Glucogenic and ketogenic 4 ketogenesis in ‘amino acids liver ‘Osmotic diuresis Np loss in urine Ketonuria Polyuria Water and electrolyte Dehydration Acidosis Coma Death Fig. 4.27 Profile in diabetes mellitus.| L Medical Biochemistry: Preparatory Manual for Undergraduates © Gestational, ile. any degree of glucose intolerance diagnosed in pregnancy © Classification © Primary © Secondary © Primary diabetes mellitus can be classified as follows: (i) Glinical includes IDDM (Type-I) and NIDDM (Type-II). (ii) Subclinical, i.e. abnormal GTT. (iii) Latent: Example is the occurrence of hyperglycaemia under stress. (iv) Potential Potentiality for diabetes due to family history. © Secondary diabetes mellitus is due to: = Pancreatic diseases such as chronic pancreatitis, pancreatectomy, hae- mochromatosis. = Insulin receptor abnormality such as congenital lipodystrophy. = Drug:induced by the use of diuretics, antibiotics, contraceptive pills and steroids. - Endocrine disorders other than those of pancreatic origin. © Treatment * In IDDM cases, insulin is given subcutaneously. * In NIDDM cases, oral hypoglycaemic drug, e.g. tolbutamide, diet control, exercise and reduction of obesity are recommended. © Complications © Atherosclerosis leading to acute myocardial infarction, gangrene in legs and cerebral stroke. * Microangiopathies leading to retino-, neuro- and nephropathy. * Susceptibility to infections and nonhealing wounds. Q58. Mention the other glucose tolerance tests. Ans. « Intravenous GTT is done when abnormalities of glucose absorption are suspected. * A cortisone test is used for detecting latent diabetes or prediabetes. Q,59. How is blood glucose measured? Ans. * In clinical laboratories, plasma glucose levels are now measured using enzy- matic methods, e.g. glucose oxidase method. Automated and semiautomated analysers are used for this purpose. © Sometimes, when blood glucose levels are required outside the laboratory, glucose reagent strips are used. These strips are impregnated with reagent which reacts with glucose and gives a colour reaction which is assessed. © Such methods are not very accurate, but can be used when rapid or frequent measurements of glucose are required. Q.60. What are the inborn errors of carbohydrate metabolism? Ans. In instances where the regulatory mechanism fails, the body starts to suffer certain diseases. They are as follows: © Hereditary defects of lactase, sucrase and a-dextrinase lead to intolerance of their corresponding substrates, e.g. lactose, sucrose and dextrins. alactosidase For example: Lactose eM, Giicose + Galactose (lactase) ‘© Lactose intolerance: Many Asians and adult blacks suffer from lactose intolerance. They are deficient in lactase. Lactose is not utilised in the body. AccumulationChapter 4 — Carbohydrate Metabolism of lactose leads to bloating, diarrhoea and dehydration. In such cases, lactose should be avoided in the diet. Malabsorption of lactose can be determined by measuring the Hy content of the patient's breath after a test dose of lactose has been consumed. * Defects in fructose metabolism: Fructose intolerance is due to the absence of enzyme, aldolase while fructosuria results due to absence of fructokinase. * Glycogen storage diseases: The causes and characteristics of the glycogen storage diseases are given in Table 4.5. © Mucopolysaccharide metabolism: Deficiency of enzymes (hydrolytic) involved in the degradation of glycosaminoglycans (GAGs) leads to certain disorders. For example, Hunter syndrome occurs due to iduronate sulphatase deficiency. * G

You might also like

• B Y: - Idr. Megha Gaur BDS I

  

  No ratings yet

  B Y: - Idr. Megha Gaur BDS I

  89 pages
• CARBOHYDRATE ABSORPTION (Ref)

  

  No ratings yet

  CARBOHYDRATE ABSORPTION (Ref)

  22 pages
• Heme Metabolism and Synthesis Pathways

  

  No ratings yet

  Heme Metabolism and Synthesis Pathways

  138 pages
• Metabolic Pathways & Regulation

  

  No ratings yet

  Metabolic Pathways & Regulation

  27 pages
• Lipoprotein Metabolism

  

  No ratings yet

  Lipoprotein Metabolism

  23 pages
• Protein Metabolism Introduction

  

  No ratings yet

  Protein Metabolism Introduction

  36 pages
• Glycogenesis: Presenters: Casilang, Michelle Anne Marina Ferrer, Julia Sophia Untalan, Jonalyn Ydeo, Revenne

  

  No ratings yet

  Glycogenesis: Presenters: Casilang, Michelle Anne Marina Ferrer, Julia Sophia Untalan, Jonalyn Ydeo, Revenne

  12 pages
• 7 - Enzymes

  

  No ratings yet

  7 - Enzymes

  15 pages
• Essentials of Biochemistry Carbohydrates Slides

  

  No ratings yet

  Essentials of Biochemistry Carbohydrates Slides

  51 pages
• Cholesterol Synthesis, Transport, and Excretion: Abdul Salam M Sofro Faculty of Medicine YARSI University

  

  No ratings yet

  Cholesterol Synthesis, Transport, and Excretion: Abdul Salam M Sofro Faculty of Medicine YARSI University

  39 pages
• Blood Formation and Fluid Regulation

  

  No ratings yet

  Blood Formation and Fluid Regulation

  56 pages
• Medical Biochemistry (Chapter 3)

  

  No ratings yet

  Medical Biochemistry (Chapter 3)

  25 pages
• Introduction To Biochemistry Carbohydrates

  

  No ratings yet

  Introduction To Biochemistry Carbohydrates

  49 pages
• 5 Enzyme Kinetics-Inhibition

  

  No ratings yet

  5 Enzyme Kinetics-Inhibition

  40 pages
• CARBOHYDRATES

  

  100% (3)

  CARBOHYDRATES

  38 pages
• Understanding the TCA Cycle Process

  

  No ratings yet

  Understanding the TCA Cycle Process

  10 pages
• Red Blood Cell Membrane Structure & Function

  

  No ratings yet

  Red Blood Cell Membrane Structure & Function

  30 pages
• Metabolism: Carbohydrate Metabolism: Inge Holsbeeks

  

  No ratings yet

  Metabolism: Carbohydrate Metabolism: Inge Holsbeeks

  89 pages
• Metabolism-1-Harper's Illustrated Biochemistry

  

  100% (1)

  Metabolism-1-Harper's Illustrated Biochemistry

  4 pages
• Glycogen Metabolism

  

  No ratings yet

  Glycogen Metabolism

  35 pages
• Body's Buffer Systems Explained

  

  No ratings yet

  Body's Buffer Systems Explained

  5 pages
• Glycolysis and Gluconeogenesis Learning Goals: 1. Learn The Reactions

  

  100% (2)

  Glycolysis and Gluconeogenesis Learning Goals: 1. Learn The Reactions

  2 pages
• Hexose Mono Phosphate (HMP) Shunt

  

  No ratings yet

  Hexose Mono Phosphate (HMP) Shunt

  28 pages
• Metabolism of Nucleoproteins Part I

  

  No ratings yet

  Metabolism of Nucleoproteins Part I

  50 pages
• Pentose Phosphate Pathway

  

  No ratings yet

  Pentose Phosphate Pathway

  4 pages
• Enzyme Action and Metabolism Overview

  

  No ratings yet

  Enzyme Action and Metabolism Overview

  43 pages
• Cholesterol Synthesis and Regulation

  

  No ratings yet

  Cholesterol Synthesis and Regulation

  38 pages
• PC-1 Students Exam 2023

  

  No ratings yet

  PC-1 Students Exam 2023

  24 pages
• Overview of Proteins and Amino Acids

  

  No ratings yet

  Overview of Proteins and Amino Acids

  35 pages
• Isoenzymes

  

  No ratings yet

  Isoenzymes

  10 pages
• Anaplerotic and Cataplerotic Reactions of The Tca Cycle

  

  No ratings yet

  Anaplerotic and Cataplerotic Reactions of The Tca Cycle

  4 pages
• Protein Metabolism Insights

  

  No ratings yet

  Protein Metabolism Insights

  35 pages
• Cori Cycle and Muscle Metabolism Explained

  

  100% (3)

  Cori Cycle and Muscle Metabolism Explained

  69 pages
• Biochemistry Test Blueprint & Review

  

  No ratings yet

  Biochemistry Test Blueprint & Review

  10 pages
• Protein Digestion & Absorption

  

  No ratings yet

  Protein Digestion & Absorption

  20 pages
• Lecture Notes 3: Biochemistry: Biology A Eaton Rapids High School K.Coppins

  

  100% (1)

  Lecture Notes 3: Biochemistry: Biology A Eaton Rapids High School K.Coppins

  31 pages
• 8 Molecular Diagnostic Tools (Protein Biomarkers)

  

  No ratings yet

  8 Molecular Diagnostic Tools (Protein Biomarkers)

  29 pages
• Bilirubin Metabolism Overview

  

  100% (2)

  Bilirubin Metabolism Overview

  11 pages
• Nucleotide Metabolism

  

  No ratings yet

  Nucleotide Metabolism

  29 pages
• Metabolic Integration in Fed State

  

  No ratings yet

  Metabolic Integration in Fed State

  40 pages
• Nonprotein Nitrogen Compounds in Renal Tests

  

  No ratings yet

  Nonprotein Nitrogen Compounds in Renal Tests

  46 pages
• Glycogen Metabolism

  

  No ratings yet

  Glycogen Metabolism

  11 pages
• Classifications of Carbohydrates Slides

  

  No ratings yet

  Classifications of Carbohydrates Slides

  32 pages
• Carbohydrate Biochemistry Lab Guide

  

  No ratings yet

  Carbohydrate Biochemistry Lab Guide

  16 pages
• SubcellularFractionation Fa15

  

  No ratings yet

  SubcellularFractionation Fa15

  25 pages
• Protein Metabolism Overview

  

  No ratings yet

  Protein Metabolism Overview

  12 pages
• Uronic Acid Pathway

  

  No ratings yet

  Uronic Acid Pathway

  15 pages
• De Novo Synthesis of Fatty Acids

  

  100% (1)

  De Novo Synthesis of Fatty Acids

  29 pages
• Hemoglobin Biosynthesis and Catabolism

  

  No ratings yet

  Hemoglobin Biosynthesis and Catabolism

  42 pages
• 41 & 42 - Nucleic Acid Metabolism

  

  100% (2)

  41 & 42 - Nucleic Acid Metabolism

  56 pages
• Gluconeogenesis Explained

  

  100% (1)

  Gluconeogenesis Explained

  11 pages
• Protein 1

  

  No ratings yet

  Protein 1

  36 pages
• Lecture 2 - Amino Acids & Protein Chemistry

  

  No ratings yet

  Lecture 2 - Amino Acids & Protein Chemistry

  12 pages
• Galactose and Fructose Metabolism

  

  No ratings yet

  Galactose and Fructose Metabolism

  11 pages
• Lipids and Lipid Metabolism

  

  No ratings yet

  Lipids and Lipid Metabolism

  81 pages
• Integration of Metabolism 2019

  

  100% (1)

  Integration of Metabolism 2019

  68 pages
• Glycolysis: Pathway and Regulation Overview

  

  No ratings yet

  Glycolysis: Pathway and Regulation Overview

  16 pages
• Carbohydrate Digestion and Absorption

  

  83% (6)

  Carbohydrate Digestion and Absorption

  31 pages
• Lecture 1

  

  No ratings yet

  Lecture 1

  63 pages
• Unit 6A Carb Digestion Absorption Metabolism

  

  No ratings yet

  Unit 6A Carb Digestion Absorption Metabolism

  117 pages