Concept of Energy in Biosystems CELLS OBTAIN ENERGY BY THE OXIDATION OF FOOD STUFFS We already know that living objects require a continuous supply of free energy mainly for the following four purposes. (i) To synthesis macro molecules from simpler and smaller precursors. (ii) To transport molecules and ions actoss membranes against gradients. (iii) To perform mechanical work, as in the muscle contraction and (iv) To ensure fidelity of information transfer. The free energy of these processes is derived from the environment ‘The phototrophs obtain this energy by trapping light energy from the sun. On the other hand, the chemotrophs obtain it by the oxidation of food stuffs. This free energy is partly transformed into a special form before it is used for biosynthesis, transport, and fidelity. ‘The special carrier of free energy is adenosine triphosphate (ATP). ATP plays a central role in the transference of free energy from the exergonic (= energy yielding) to the endergonic (= energy requiring) processes in the cells. During breakdown of energy rich food stuffs or fuel molecules, some of the free energy is harnessed to make ATP from adenosine diphosphate (ADP) and inorganic phosphate (Pi), a process that requires input of free energy. ATP then donates much of its chemical energy to energy requiring processes (biosynthesis, transport etc.) by undergoing a breakdown to ADP and Pi. ‘Adenosine triphosphate (ATP) was found to be present in all types of cells- animal, plant and microbial. Although, first thought to be concerned with muscular contraction, only ATP has now been assigned many cell activities.Ka e MODERN COLLEGE CHEMISTRY (SEMESTER-V)’ ATP Musoular CO Is Active Genetic formation contraction Energy yielding transport Trapsfer oxidation of fuel molecules Fi The ATP Cycle in Cells Fritz A. Lipmann postulated that ATP is the primary and universal carrier of chemical energy in cells. He adequately stated that the small chemical units from which the material of organisms is built are glued together by an enormously versatile condensing reagent ATP”. He also first proposed the ATP cycle shown in a present form in figure 5.1. Adenosine triphosphate (ATP) and its successive hydrolysis products, adenosine diphosphate (ADP) and adenosine monophosphate (AMP) are nucleotides, consisting of an adenine, a ribose and a 3, 2 or 1 phosphate groups (s) respectively (Fig, 5.2). Phosphoanhydride yy ‘bonds —Phosphoester bonds 1 1 1 1 Trafic “O—P: = O--P=-O--CHp 0, i 1 1 ' i 1 1 1 a 1 ' I ' 1 t 1 ' Adenosine i Orr 1 AMP. Fig. 5.2. The structure of ATP, Indicating its relationship to ADP, AMP and adenosine. ATP, ADP and AMP occur not only in cell cytosol but also in mitochondria and the nucleus. In normal respiring cells, ATP make up about 70% or more of the sum of all three ademine ribonucleotides.\CONCEPT OF ENERGY IN BIOSYSTEMS EE ATP serves as the principal immediate donor of free energy in biological systems rather than as a storage of energy. In a typical cell, an ATP molecule is consumed within a minute of its formation. The turn over of ATP is very high. For example, a resting human consumes about 40 kg ATP in a day. During strenuous labour, the ATP is consumed at the rate of even 0.5 kg per minute. The endergonic processes such as biosynthesis, active transport etc. can occur only if ATP is continuously regenerated from ADP. Phototrophs harvest the free energy in light to regenerate ATP whereas chemotrophs form ATP by the oxidation of food stuffs. Hence ATP acts as universal currency of free energy in biological systems. METABOLISM This term applies to the assembly of biochemical reactions which are employed by the organisms for the synthesis of cell materials and for the utilisation of energy from their environments. Alternatively, metabolism of an organism or a cell is the sum total of all the enzyme catalysed reactions that occur * in an organism or a cell. The large number of reactions in a cell are organised into a relatively small number of pathways. It is a highly co-ordinated and purposeful cell activity in which multienzyme systems cooperate. For the magnitude of metabolism, we take the examples of micro-organisms and humans. 1. Bacteria (microorganism), ¢.g.; Escherichia coli can double in number every 40 minutes in a culture medium containing glucose and inorganic salts. The components of the medium are depleted and very little is added to the medium by the cells. Each cell contains hundreds to thousands of molecules of each of about 2500 different proteins and 1000 types of organic compounds and a variety of nucleic acids. Clearly, the bacterial cells participate in a variety of metabolic activities in a remarkable way. 2. Humans (adults) maintain a constant weight for about 40 years. In this period, a total of about 60 quintals of solid food and 45000 litres of water are metabolised. But still the body weight and body composition remain almost constant. In all cells, metabolism enables the cell to perform its vital functions. It performs the following specific functions. (i) to obtain chemical energy from the degradation of energy rich nutrients or from captured solar energy. (ii) to convert nutrient molecules into precursors of cell macromolecules. (iii) to assemble these precursors into proteins, lipids, polysaccharides, nucleic acids and other cell components. (iv) to form and degrade biomolecules required in specialised functions of cells. These features of metabolism are closely inter-related since the synthesis of the molecules, that are component of cell, requires an input of energy, while at the same time, it is obvious that the cell components are needed to provide the energy supply to control intracellular solute concentrations. These components makeup the cell membrane and its constituent transport proteins. The specialised functions also require biosynthetic processes as well as supply of energy. Catabolism refers to the pathways breaking down food materials into simpler compounds and resulting in the release of energy contained in them. The process of anabolism utilise food materials and energy to synthesis cell components.EK MODERN COLLEGE CHEMISTRY (SEMESTER-V) Functions of Metabolism The enzyme catalysed chemical reactions, that bring about transformations of en ee compounds vital to the organism, constitute metabolic pathways. The mmolernies or d a oe ds which participate in these reactions are called metabolic intermediates or metabolites. a ea esis of cell constituents begins with the metabolic pathways which supply the building bloc ra 1 major cell constituents are complex carbohydrates, the proteins, the lipids and the nucleic acids. They are formed from relatively simple starting materials. In mammalian cells, these key metabolites come from one of the following three sources. (@) Absorption of the metabolites from outside the cell and by implication, if we consider the organism as a whole, by absorption of the metabolites as dietary constituents from the alimentary tract. (i) Release of the metabolite from a source stored in the cell. The metabolite may be released either from the molecule used solely for storage (such as glycogen) or from a molecule, that has another function within the cell. (iii) The metabolite may be formed by the metabolism of a simpler precursor. However, the Precursor must be absorbed by the cell or derived from a source stored within the cell. Following are the key functions of the metabolism. (@) Metabolism enables the cell to convert same of the energy found in the nutrients into a form which will support biosynthesis and maintenance of cell’s other energy requiring processes. (ii) Biosynthetic reaction pathways in the cell require an input of energy and this will normally be in the form of ATP or a reduced co-enzyme (NADH*, NADPH*). It is noteworthy that the usefulness of ATP (Adenosine triphosphate) as an energy source is not a consequence of any special or high energy form of the phosphate bond,but is merely a function of how far the hydrolysis of ATP is displaced from equilibrium. Metabolism cells have five reaction schemes which are capable of supplying cells with adequate amounts of energy in one of these forms. These energy conserving pathways are described below: 1. The glycolytic pathway: This converts glucose into pyruvate or lactate. 2. The citric acid cycle : This converts acetate units into carbon dioxide. 3. The pentose phosphate pathway : This converts glucose-6-phosphate into pentose phosphate and reduces NADP coenzyme. 4. B-oxidation of fatty acids : This converts fatty acids into CO2 and also reduces the coenzymes. 5. Oxidation phosphorylation : This is the electron transport phosphorylation process in which molecular oxygen is used to oxidise the coenzymes, which are reduced in the other four pathways, with the production of water and the conversion of ADP plus phosphate into ATP. Metabolism provides key metabolites which are required for the synthesis of many essential cell components. In addition to supplying the cell with energy, these central pathways provide precursors required for the formation of carbohydrates, proteins, lipids and nucleic acids which the cell is "NADH stands for Nicotinamide adenine dinucleotide + NADPH stands for Nicotinamide adenine dinuclotide phosphate‘CONCEPT OF ENERGY IN BIOSYSTEMS cae composed of. Most of the pathways are found in almost all mammalian cells, although, some reactions which occur in specialised cells have been incorporated for the sake of completeness e.g.; the metabolism of galactose and fructose, the formation of urea and glycogen and the hydrolysis of glucose-6-phosphate. Glycolysis, which is responsible for the conversion of glucose into 3-carbon compounds, illustrates the role of these metabolic pathways. The glycolytic pathway supplies the cell with ATP and also provides the acetyl-coenzyme-A and glycerol-3-phosphate required to form the phospholipids needed for the cell membranes. The pentose phosphate pathway provides a different sort of example since it is responsible for supplying energy (as (NADPH) for fatty acid synthesis, but it also supplies the phosphoribosyl-1-pyrophosphate needed for the formulation of nucleotides, the precursors of nucleic acid. Catabolism and Anabolism (a) Catabolism. It is the conversion of complex organic substances into more simple compounds (CO> + HO) by living cells. It is the destruction or the breaking down phase of metabolism. Such reactions may field energy in the form of ATP. Catabolism involves such processes as glycolysis, glycogenolysis, proteolysis and oxidation of fatty acids (6) Anabolism. It is a part of constructive metabolism and involves the synthesis of living things of complex molecules from simple ones in the cell. For example, proteins, nucleic acids, polysaccharides and other polymers from their manomers presursors. The energy of these processes is derived from hydrolysis of ATP. Anabolism includes the process of building of body tissues from simple materials supplied by the production of food digestion. CATABOLIC PATHWAYS OF FATS PROTEINS AND CARBOHYDRATES Catabolic process involves the conversion of complex organic substances into more simple compounds i.e. carbon dioxide and water by living cells. A. Metabolic pathways of Proteins It involves protein metabolism and involves the general aspects of amino acid degradation. (a) Aminoacid degration. Amino acid degradation forms major metabolic intermediates that can be converted into glucose or be oxidised by citric acid cycle. Based on their catabolic products, amino acids are classified as glucogenic or ketogenic. (i) Glucogenic are those amino acids that generate precursors of glucose (pyruvate or citric acid cycle intermediate). Examples are Ala, Arg, Asp, Asn, Cys, Glu, Gin, Gly, Hist, Met, Pro, Ser, Thr and Val. (ii) Ketogenic are those aminoacids that are degraded to acetyl-CoA and acetoacetyl-CoA because they give rise to ketone bodies. Lys, Phe, lie, Trp and Tyr are both glucogenic and ketogenic. Table 5.1 Classification of Amino acids. Glucogenic Katogenic | Both glucoganic and katogenic Alanine, Arginine, Asparagine, Isoleucine, Lysine, Phenylalanine, Aspartate, Cysteine, Glutamate, Tryptophan, Tyrosine. Glutamine, Glycine, Histidine, Methionine, Proline, Serine, Threonine, Valinesz | MODERN COLLEGE CHEMISTRY (SEME It is important to note that nearly all the aminoacids yield on degradation cither an intermediate of the citric acid cycle or pyruvate or acetyl-CoA. Leu, Lys, Phe, Trp and Tyr degrade to give acetoacetic acid, Table 5.2 End products of Amino acid metabolism. Final product Amino acid (s) Glycine, Alanine, Serine, Cystine and Threonine (2) Leucine (2) Lysine (4), Leucine (4), Phenyl alanine (4) Tryptophan (4) and Tyrosine (4) Arginine (5), Glutamic acid, Glutamine, Histadine (5) and Proline Isoleucine (4), Methionine and Valine (4) Phenylalanine (4) and Tyrosine (4) Asparagine and Aspartic acid Pyruvic acid Acetyl-CoA Acetaocetic acid (or its CoA ester) a-ketoglutaric acid Succinyl-CoA Fumarate Oxaloacetic acid 1. Ten Amino acids are degraded to Acetyl-CoA. The carbon skeletons of ten amino acids yield acetyl-CoA which enter the citric acid cycle directly. Out of these, five are degraded to acetyl-CoA via Pyruvate. These five are alanine, cysteine, glycine, serine and tryptophan. The other five amino acids are converted into acetyl CoA and or acetoacetyl-CoA which then cleave to acetyl-CoA. These amino acids i.e., Leucine, Lysine, Phenylalanine, Tryptophan and Tyrosine come under this class. (i) Alanine gives pyruvate directly on transamination with a-ketoglutarate, ii) The side chain of tryptophan is broken to give Alanine and thus pyruvate. (ii) Cysteine is converted to pyruvate by removal of sulphur atom and then involves transamination. (iy) Serine is deaminated to pyruvate by serine dehydratase and so on. 2. Five Amino acids are converted into a-ketoglutarate. The carbon skeletons of arginine, histidine, proline, glutamate, glutamine enter the citric acid cycle at a-keto glutarate. Out of these, proline, glutamate and glutamine have five carbon sheletons. The cyclic structure of proline is opened by oxidation of the carbon farthest from the carboxyl group to create schiff s base and the hydrolysis of the Schiff s base to a linear semialdehyde. glutamate-y-semialdehyde. It gets further oxidised at the same carbon to give glutamate, Deamination of glutamate produces the citric acid cycle intermediate. 3. Four Aminoacids are converted into Succinyl-CoA. The carbon skeletons of methionine, Isoleucine, threonine and valine are degraded by pathways that produce succinyl-CoA. Methyl malonyl-CoA is an intermediate in the break down of these four amino acids, Methionine donates its methyl group to one of the many possible acceptors through S-adenosyl methionine and three of the four remaining atoms of its carbon skeleton are converted into those of propionate as propionyl-CoA. 4. Three branched chain Amino acids are degraded in extrahepatic tissues. We know that most of the amino acids are mainly catabolised in liver, the three aminoacids with branched side chains, viz. leucine, isoleucine and valine are oxidised as fuels Primary in the extrahepatic tissues such as muscle, adipose, kidney and brain tissue. These amino acids share the enzymes amino transferase and branched chain a-keto dehydrogenase to form acyl-CoA. 5. Two Aminoacids are degraded to Oxaloacetate. The carbon skeletons of Asparagine and Aspartate finally enter the citric acid cycle via oxaloacetate. Asparagine is hydrolysed by the enzyme(CONCEPT OF ENERGY IN BrosvsteMs oe asparaginase to NH and aspartate. Asparatate then undergoes transamination reaction with a a-keto glutarate to yield glutamate and oxaloacetate. Hence we see that all protein amino acids lose their N-atoms and then degrade through dehydrogenation, decarboxylation and other reactions to give portions of their carbon skeleton in the form of five central metabolites that can enter citric acid cycle. In citric acid cycle, they are fully oxidised to carbon dioxide and water. It may be noted that ATP is generated by oxidative phosphorylation and hence amino acids centribute to the total energy supply of the organisms. () Deamination. It is an enzyme catalysed process in which an amino (-NH;) group is removed from an amino acid or other amine by the action of deaminating enzymes. A number of nucleotide bases undergo spontaneous loss of their exocyclic amino group (deamination). For example, under typical cellular conditions, deamination of cytosine (in DNA) to uracil occurs in about one of every 10’ cytidine residues in 24 hours. This corresponds to about 100 spontaneous events per day in a mammalian cell. Deamination of adenine and guanine occurs at about 1/100th this rate. The slow cytosine deamination reaction seems innocuous but almost certainly the reason why DNA contains thymine rather than uracil. The product of cytosine deamination (Uracil) is readily recognised as foreign in DNA and is removed by a repair system. Some deamination processes are shown below : NH2 He Na HN’ ns HN ae) AIL} LI JD N N Ny I I | Cytosine Uracil 5-Methyicytosine Thymine NHo 9 9 e N, N H na \ Hl SS. «tit iN uN S ly lL : dy | 2s NTN WNT tan SN of SN NN | I Hi HI Adenine Hypoxanthine Guanine Xanthine If DNA normally contained uracil, recognization of uracils resulting from cytosine deamination would be more difficult and unrepaired. Uracils would lead to permanent sequence changes-as they were paired with adenines during replication. It may be noted that cytosine deamination would gradually lead to a decrease in G = C base pairs and an increase in A = U base pairs in the DNA of all cells. Over the millenia, cytasine deamination could eliminate G = C pairs and the genetic code that depend on them. ‘An important reaction in deoxyribonucleotides is the hydrolysis of the N-B-glycosyl bond between the base and the pentose. This occurs at a higher rate for purines than for pyrimidines. As many as one in 10° purines are lost from DNA every 24 hours under typical cellular conditions. (c) Transamination. The first step in the catabolism of most L-amino acids (once they have reached the liver), is removal of the a-amino groups, promoted by enzymes, called aminotransferases or transaminases. In these transamination reactions, the a-amino group is transferred to the a-carbon atom of a-ketoglutarate, leaving behind the corresponding a-keto acid analog of the amino acid. There is no net examination (loss of amino groups) in these reactions, because the @-ketoglutarate becomes aminated as the a-amino acid is deaminated.MODERN COLLEGE CHEMISTRY (SEMESTER The effect of transamination reactions is to collect the coo~ coo amino groups from many different amino acids in the form | of L-glutamate. The glutamate then functions as an amino ie HeN=—c_y group donor for biosynthetic pathways or for excretion Mes by pathways that lead to the elimination of nitrogenous waste | | products. Cells contain different types of amino transferases. CHp CH Many are specific for q-ketoglutarate as the amino group bo ea acceptor but differ in their specificity for the L-amino acid. oon ae The reactions catalysed by aminotransferases are freely reversible having equilibrium constant, almost equal to 1. All amino transferases have the same prosthetic group and the same reaction mechanism. The prosthetic group is pyridoxal phosphate (PLP), the co-enzyme form of pyridoxamine vitamin in Bg. Pyridoxal phosphate functions as an intermediate carrier of amino groups at the active site of amino transferases. It undergoes reversible transformations between its aldehyde form, pyridoxal phosphate, which can accept an amino group and its aminated form, pyridoxamine — L-Amino acid mea phosphate, which can donate its amino group to an a-ketoacid. Thus, the incoming amino acid binds to the active site, donates its amino group to pyridoxal phosphate and departs in the form of an a-keto acid. The incoming a-ketoacid then binds, accepts the amino group from pyridoxamine phosphate and departs in the form of an aminoacid. B. Metabolic pathways of Facts (Fatty acid metabolism) The oxidation of long chain fatty acids to acetyl-CoA is an energy yielding pathway in many organisms and tissues. In mammalian heart and liver, it provides about 80% of the energetic needs under all physiological circumstances. The electrons removed from fatty acids during oxidation pass through the respiratory chain, driving ATP synthesis, The acetyl-CoA produced from fatty acids may be completely oxidised to CO) in the citric acid cycle resulting in further energy conservation. In liver, acetyl Co A may be converted to ketone bodies, water soluble fuels exported to the brain and other tissues when glucose is not available. In higher plants, acetyl-CoA serves primarily as biosynthetic precursor and only secondarily as fuel. Mitochondrial oxidation of fatty acids takes place in three steps : (i) The long chain acid is oxidised to yield acetyl residues in the form of acetyl - CoA . (ii) The acetyl residues are oxidised to CO} via, the citric acid cycle. (iii) Electrons derived from the oxidations in step (i and step (ii) are passed to 2 vid the mitochondrial respiratory chain, providing the energy for ATP synthesis by oxidative phosphorylation. B-oxidation pathway. First Stage. In A-oxidation pathway, the fatty acids undergo oxidative removal of successi¥? two carbon units in the form of acetyl-CoA; starting from the carboxyl end of the fatty acy! chai For example, palmitic acid (16 carbons) undergoes seven passes through this oxidative sequen each pass, two carbon atoms are lost as acetyl-CoA. After seven cycles, the last two carbons left as acetyl CoA. Thus, the overall result is the conversion of 16-carbon palmitic acid to 8 carbon acetyl-CoA molecules. The formation of each acetyl-CoA molecule requires the remove! four hydrogen atoms from the fatty acyl moiety by the action of dehydrogenases.‘CONCEPT OF ENERGY IN BIOSYSTEMS en ees In this stage of fatty acid oxidation, the acetyl residues of acetyl-CoA are cose 19 2 ia clic acid cycle, Acetyl-CoA derived from fatty acid oxidation enter a common y to ion alongwith acetyl-CoA derived from glucose (via glycolysis and pyruvate oxidation). ae hird Soe First stage and second stage of fatty acid oxidation produce the electron carriers, \ Hp. These in the third stage donate electrons to mitochondrial respiratory chain and el ectrons are carried to oxygen. This flow of electrons is coupled with phosphorylation of ADP to ATP. Thus, the energy released as a result of fatty acid oxidation is conserved as ATP. ‘The A-oxidation of saturated fatty acid has four basic steps. First step : a, B-dehydrogenation of Acyl -CoA. Four enzyme catalysed reactions make up the first stage of fatty acid oxidation. First step includes the oxidation of acyl-CoA by an acyl-CoA dehydrogenase to produce an enoyl-CoA with a trans double bond between a, and f-carbon atoms. I i RCH, - CH, — CH» - C-S-CoA+FAD —+ RCH, ~ CH = CH- C-S~CoA+FADH, Trans-A?-enoyl-CoA AG=-4.8 kcal mol’. Three acyl-CoA dehydrogenases are found in the matrix of mitochondria. They all have FAD as a prosthetic group. FAD is Flavin adenine denucleotide. It is a riboflavin derivative that functions in oxidation-reduction reactions. Second Step. Hydration of a, f-unsaturated acyl-CoA. Here a mole of water is added to the double bond of trans-A?-enoyl-CoA to form L-stereo isomer of -hyroxyacyl-CoA. The reaction is catalysed by enoyl-CoA hydratase. It has broad specificity with respect to the length of the acyl group. ° ° Ml i R-CH, -CH=CH-C-S-CoA+H,0 ==> R-CH, -CH-CH-C-S-CoA | 1 OH H Lis Bhydroxyl acyl-CoA Third Step. Oxidation of b-hydroxy acyl-CoA. Here L-B-hydroxyl acyl-CoA is dehydrogenated to form f-ketoacyl-CoA by the action of an enzyme, f-hydroxy acyl-CoA dehydrogenase. It is specific for the L-stereo isomer of the hydroxyacyl substrate. NAD+ is the electron acceptor in this reaction and the NADH thus formed, donates its electrons to NADH dehydrogenase. a I RCH -CH-CH-C-S-CoA+NAD' <== RCH, - CO CH, -C-S-CoA+NaDH+H" rot f-ketoacy-CoA OH H AG = +3.75 kcal mol”. Fourth Step. Thiolysis. Thiolysis is the splitting by thiol (SH-) group aided by enzymatic catalysis. This step brings about the cleavage of -ketoacyl-CoA by the thiol group of a second mole of CoA which gives acetyl-CoA and an acyl-CoA shortened by two carbon atoms, The thiolytic cleavage is catalysed by the enzyme acyl-CoA acetyl transferase which has broad specificity. The shortening of a fatty acyl-CoA derivative by two carbon atoms can be represented by the equation. i i RCH, -CO-CH, - C-S-CoA+Enz-SH <==» RCH,CO-S—Enz +CH;—C-S—CoA B-ketoacy!-CoA Ue) ‘Acyl Enz AcetyCoAMODERN COLLEGE CHEMISTRY (SEMESTER-V; ~Co-S-CoA+Enz-SH Acyl-CoA By repeated turns of the cycle, a fatty acid is degraded to acetyl-CoA molecules with one being produced every tum till in the last cycle, two are produced. R-CHp -Co-S-Enz+CoA-SH === RCH t R—CH2—CH2—C—S—CoA (Acyl-CoA) 9° FAD I ‘Acyl-CoA dehydrogenase R—C—S—CoA DH i CHs —C —S—CoA q (Acetyi-CoA) trans R—CH=CH—C—S—CoA Thiolase (6-Enoyl-CoA) CoA- SH: EnoyCoA J t1.0 hydratase ° a t 1 R I CHp—C—S—CoA R—CHOH —CH2—C—S—CoA (@-Ketoacyl-CoA) (B-Hydroxyacyl-CoA) NADH + H* NAD (6-Hydroxyacyl-CoA) dehydrogenase Fig. 5.3. The f-oxidation cycle for fatty acids. C. Carbohydrate metabolism-Glycolysis Glycolysis is an enzymatic breakdown of glucose to Pyruvic acid. In aerobic respiration, glycolysis yields pyruvic acid at the Kreb’s cycle while in anaerobic respiration of carbohydrates, the pyruvic acid is converted into lactic acid. The major function of carbohydrates in metabolism is as a fuel to be oxidised and provide energy for other metabolic purposes. The carbohydrate is utilised by cells mainly as glucose. The three principal monosaccharides resulting from digestive processes are glucose, fructose and galactose. Glucose is also produced from other dietary components by the liver. Both fructose and galactose are easily converted to glucose by the liver. The break down of glucose molecule occurs in a series of enzyme catalysed reactions to yield two molecules of the three carbon compound pyruvate. During the sequential reactions of glycolysis, some of the free energy released from glucose is converted in the form of ATP and NADH. Fermentation is a general term for the anaerobic degradation of glucose or other organic nutrients to obtain energy, conserved as ATP, Because living organisms first arose in an atmosphere without oxygen, anaerobic breakdown of glucose is probably the most ancient biological mechanism for obtaining energy from organic fuel molecules, The breakdown of the six carbon glucose into two molecules of the three carbon pyruvate occurs in teji steps. The first five steps constitute the preparatory phase, (@ Preparatory phase. The various steps involved in this phase are ; (i) Glucose is first phosphorylated at the hydroxyl group on C - 6,(ii) The D-glucose-6-phosphate thus formed is converted to D-fructose- phosphate. (iii) The product in step (ii) is again phosphorylated and this time at C-I to give D-fractose -1, 6-bisphosphate. For both phosphorylations, ATP is the phosphoryl group donor. As all sugar derivatives in giycolysis are the D-Isomers, we usually omit the D-designation. (iv) Fructose-1, 6-biphosphate is split to yield two 3-carbon molecules, dihydroxy acetone phosphate and glyceraldehyde 3-phosphate. This is the ‘lysis’ step that gives the pathway its name. (v) The dihydroxy acetone phosphate is isomerised to a second molecule of glyceraldehyde-3-phosphate, This ends the first phase of glycolysis. It may be noted that two molecules of ATP are invested before the cleavage of glucose into 3-carbon pieces. Glucose 7 Preparatory phase HO—CH2 ¥ O. Phosphorylation of glucose Tes ne Wa and its conversion to priming 0 ‘Kon n/t glyceraldehyde 3-phosphate reaction ADP HONS "oH HOH Glucose 6-phosphate (i) Hexokinase @®—oO-—CH2 H Q (i) Phosphohexose isomerase a) H HONG Yon (ii) Phospho-fructokinase-1 on (Wv) Aldolase Fructose 6—-phosphate @®—O-—CH, CHe—OH _(v) Trlose phosphate isomerase ‘ATP J m priming mo C H\H-HO/oH reaction ADP yi OH oH Fructose 1,6-phosphate Cesvage @—0— chy -O\_ He -0-® of 6-carbon sugar phosphate to} ") H\H —-HO/oH two carbon oun OH oH phosphates ° O Glyceraldenyde phosphate © ©—O—CHz—CH—C\ IH + Dihydroxyacetone phosphate 0 cir 9 OneoH °glycolysis, the energy of ATP is invested, raising the free In short, in the preparatory phase of hs all the metabolised hexoses are energy content of the intermediates and the carbon chains of. converted into a common product, glyceraldehyde-3-phosphate. (b) Pay off phase. In the pay off phase of glycolysis, the energy gain occurs. It involves the follows steps : (vi) Each molecule of glyceraldehyde-3 -phosphate is oxidised and phosphorylated by inorganic phosphate to form 1, 3-biphosphoglycerate. (vii) Energy is then released as the two molecules of 1, 3-bisphosphoglycerate are converted to two molecules of pyruvate through further steps, (viii), (ix) and (x) Much of this energy is conserved by the coupled phosphorylation of four molecules of ADP to ATP. The net Oo Zz Giyceraldehyde 3-phosphate. @®-—o0—cHz— f= q& x oH Dihydroxyacetone phsophate O—CHz— C — CH20H . @-0- ot — oe © Pay off phase Oxidative conversion of Glyceraldehyde 3 — phosphate (2) _ glyceraldehyde 3-phosphate to Q-o- tHe ot Kets come wi 2P; OH formation of ATP and NADH vi M-enap* oxidation and | phosphorylation |} (vi) Glyceraldehyde 2.NADH + Ht ‘3-phosphate dehydrogenase 1, S-Biephoephoglcerate (2) (vit) Phosphoglycerate kinase first aTP- (i)|| > app forming reaction (vil) Phosphoglycerate (substrate-tevel oP mutase shosphorylati eee Z (x) Enolase hosph O—CHz— CH— < '3-Phosphoglycerate (2) ®- if o ns OH kinase (viii) ye) CHe— cH a 2-Phosphoglycerate (2) OH | oO ® oe NK 2H20 Phosphoenolpyruvate (2) second ATP— 2ADP forming reaction x (substrate-level phosphorylation) za CHe—o— Pyruvate (2)yield is two molecules of ATP per molecule of glucose used, because two molecules of ATP are invested in the preparatory phase, Energy is also conversed in the pay off phase in the formation of two molecules of NADH per molecule of glucose, In glycolysis, the important transformations are : (i) degradation of the carbon skeleton of glucose to yield pyruvate. (ii) phosphorylation of ADP to ATP by high energy phosphate compounds formed during glycolysis and (iii) transfer of hydride ion to NAD+, forming NADH. FATES OF PYRUVATE We know that pyruvate is formed by glycolysis. Glycolysis is further metabolised via cne of the three catabolic routes. In aerobic organisms or tissues (under aerobic conditions), glycolysis is only the first stage m the complete degradation of glucose. Pyruvate is oxidised, with loss of its carboxyl group as carbon dioxide, to form the acetyl group of acetyl co-enzyme A. The acetyl group is then oxidised completely to carbon dioxide by citric acid cycle. The electrons from these oxidations are passed to oxygen through a chain of carriers in the mitochondrion, to form water. The energy from the electron transfer reactions drives the synthesis of ATP in the mitochondrion. The second route for pyruvate is its reduction to lactate, via lactic acid fermentation. When vigorously contracting skeletal muscle must function under low oxygen conditions, NADH cannot be reoxidised to NAD*, but NAD* is required as an electron acceptor for the further oxidation of pyruvate. Under these conditions, pyruvate is reduced to lactate accepting electrons from NADH and thereby regenerating the NAD+ necessary for glycolysis to continue. Certain tissues and cell types convert glucose to lactate even under aerobic conditions and lactate is also the product of glycolysis under anaerobic conditions in some micro organisms. The third major route of catabolism forms ethanol. In some plant tissues and micro-organisms such as brewer’s yeast, pyruvate is converted into ethanol and carbon dioxide and the process is called ethanol fermentation. It is important to note that the oxidation of pyruvate is an important catabolic process but pyruvate has anabolic fates as well. It can provide the carbon skeleton for the synthesis of amino acid, alanine. CALORIFIC VALUE OF FOOD Most common chemical reactions used to produce heat are combustion reactions. The energy teleased when a fuel or food is burnt is known as its fuel value or calorific value. It is defined as : The amount of heat energy released by the complete combustion of one gram of the fuel. Itis common to report calorific values without their associated negative sign because all combustion reactions are exothermic. Furthermore, calorific values are reported on a gram rather than mole basis because fuels and foods are usually mixtures, These do not have any fixed molecular weight. Thermochemical equation for combustion of octane, CgHyg, @ component of gasoline is : 2CgHig (I) + 2502(g) —> 16CO2(g) + 18H30 (1) AH = - 10,920 2x 114 «, + 10,920 Therefore, calorific value of CgHig (I) is = 74 = 47.9 kJ [. Molecular weight of CgHig is 114]Calorific values of some important foods and fuels are given in table 5.3. Table 5.3 Caloritic values of some common foods and fuels. f Milk Wood : Charcoal Meat | Honey Kerosene i Methane Butter . Ghee Hydrogen Among the foods, carbohydrates are the principal sources of energy due to their high calorific value. However, the calorific values of proteins is quite low. The other constituents of our food such as vitamins, salts, etc. do not provide much energy. Proteins are used by body mainly as building materials for construction of organ walls, skin, hair etc. Vitamins also play an important role in life processes. Thus, a balanced diet must contain all the major constituents-carbohydrates, fats, proteins and a small amount of vitamins. The amount of energy, the body requires, varies considerably depending upon such factors as body weight, age and muscular activity. The average adult requires about 2500-3000 kcal (10,000- 13,000 kJ) a day. This is about the same amount of energy as that consumed by a 100-watt bulb operating for a 24 hour period. Among the fuels, hydrogen has the highest calorific value. However, due to some technical problems such as storage and transportation, it is not commonly used as domestic or industrial fuel. A pescaiptive Questions ee 1, How will you say that cells obtain energy by the oxidation of food stuffs ? 2. Comment on the statement that ATP acts as a universal currency of free energy in biological systems. 3. Give a detailed description of Metabolism. 4. Write a brief account of catabolism and Anabolism, 5. Write a few lines on the functions of Metabolism. 6. Write a note on the calorification value of food. 7. Give a detailed description on the catabolic pathways of fats, 8, Write an explanatory note on the metabolic pathways of carbohydrates. 9. Explain metabolism in proteins, 10. Write notes on the following : (a) Oxidation of food stuffs and cellular energy. (b) Catabolism and Anabolism (c) Metabolic pathways of carbohydrate.