MODULE 6 CELLULAR METABOLISM

OBJECTIVES: At the end of the lesson, the students should be able to: 1. 2. 3. 4. Explain the role of energy in life. Describe the structure of ATP molecule Explain the role of ATP in life processes. Describe the major events of glycolysis and aerobic respiration.

Lesson 6.1 ATP and Energy Transfer

Almost all of our daily activities require energy and energy conversions. Energy conversions involve the release of energy stored in foods. This energy storage molecule used by most organisms is called adenosine triphosphate or ATP. Molecules of ATP transfer much of the energy needed by organisms for life processes. If electricity can supply power for many different types of appliances, ATP also do the same job. It provides energy required by many diffferent types of chemical reactions in the biological system.

CHEMICAL ENERGY AND FOOD How much stored energy is actually present in food? One gram of sugar glucose (C6H12O6) , when burned in the presence of oxygen, releases 3,811 calories of heat energy. A calorie is the amount of energy needed to raise the temperature of 1 gram of water 1 degree Celsius. The calorie ( capital C ) that is reflected in food labels is actually kilocalorie, or 1000 calories. Kilocalorie is the amount of heat energy needed to raise the temperature of 1 kg of water by 1 degree Celsius. A piece of apple pie contains about 365 Calories . The slice of pie contains about as much energy as 313 gram of the explosive TNT ( trinitrotoluene ).

ENERGY STORAGE AND RELEASE

Energy released from the breakdown of food is stored in the molecules of ATP. The energy in ATP is used for all the organisms activities. A molecule of ATP has three major parts: ribose ( pentose sugar), adenine and phosphate groups. Ribose is a five carbon sugar molecule and adenine is a nitrogen-containing base present in a nucleotide of RNA and DNA. The ribose sugar is linked to adenine and forms a nucleoside molecule called adenosine. The three phosphates groups are linked to the adenosine molecule as shown in figure 6.1 The bonds that connect the three phosphate groups contain great amount of energy. When these bonds are broken, the energy stored in the bonds is transferred to other chemical reactions. The transfer of this energy, fuels most biochemical reactions. Energy is stored and released for cell use through, a series of reactions that creates and breaks down molecules of ATP. Energy is released when an enzyme called ATPase breaks the end phosphate of an ATP molecule. The release of phosphate from ATP forms a molecule called adenosine diphosphate or ADP, plus a free phosphate. The formation of ADP from ATP releases the energy for the cellular activities that support life. However, the ADP molecules are converted back to ATP when free phosphate and energy re-form molecules of ATP. This re- formation of ATP occurs at a very fast pace approximately 10,000,000 times per second in each cell. Figure 6.1 Structure of ATP

Lesson 6.2

Glycolyis and Aerobic Respiration

Performing your daily activities without food will make you weak. But how does the food you eat converted to energy, so that you and your cells can do work. We say that our cells burn the foods that we eat. This means that the foods that you eat are oxidized or they react with oxygen converting the complex molecules of food to simple molecules of carbon dioxide and water with accompanying released of energy.. . The process by which food molecules are broken down to release energy is called cellular respiration. The different foodstuffs follow a specific chemical pathway in order to convert the chemical energy stored into a molecule of ATP. For example, the chemical pathway followed by glucose is different from the chemical pathway of amino acids and fatty acids. Cellular Metabolism Cells have to carry out tens of thousands of chemical reactions to maintain their proper function. This is known as metabolism. All of these reactions need energy, but where does the energy come from? Since the beginning of evolution, cells have developed highly efficient metabolic systems. The specific mechanisms vary in different cell types. For example, plant cells derive their energy from photosynthesis ( anabolism) and animal cells rely on aerobic respiration to fulfill their energy needs.(catabolism). Although the mechanisms differ, the end goal of all metabolic systems is the same, to produce ATP molecules.

Cellular Respiration Cells, like all things, need energy to operate. CD players use batteries for energy, cells use adenosine triphosphate (ATP) for energy. Unlike CD players, cells can produce their own energy. People don't eat ATP, though. Their food consists mostly of proteins, lipids, and polysaccharides (sugars) which need to be broken down into ATP.

Cellular respiration may be classified into two types depending on the need for oxygen. Aerobic respiration takes place in the presence of oxygen ( oxygen-dependent) while anaerobic respiration occurs in the absence of oxygen ( oxygen-independent ). The basic chemical reaction taking place during cellular respiration are the ff. 1. the consumption of ATP to provide the energy needed to start the breakdown of food. 2. dehydrogenation or the removal of hydrogen from the food molecules.

There are three major stages in converting food into ATP: food breakdown, glycolysis, and the Kreb Cycle.

Stage 1: Food Breakdown Before food can be converted into ATP, it must be broken down into simpler forms of sugar, lipid, or amino acids. Stage 2: Glycolysis The simple molecules from stage 1 must be converted into a intermediate product before it can be converted into ATP. Stage 3: Aerobic Respiration In this step, food is finally converted into ATP. Fermentation In the absence of oxygen, cells undergo fermentation to produce ATP.

The three major stages of cellular respiration is shown in the diagram below.

Stage 1: Food Breakdown

Before food can be processed into ATP, large polymeric molecules must be broken down into their basic units. For example, proteins must be broken down into amino acids and polysaccharides must be broken down into glucose. This step is known as digestion. Most of digestion takes place in the digestive tract of animals and food is broken down by secreted enzymes. Even after all of the molecules have been broken down into their basic units, most of the non-glucose molecules are subsequently converted into glucose so that they can be processed during glycolysis. The following table lists the food molecules and there basic unit. Complex Food Molecule Proteins Polysaccharides Fats (Lipids) Basic Units Amino Acids Simple Sugars (e.g. glucose) Fatty Acids and Glycerol

Stage 2: Glycolysis Glycolysis (glyco = sugar; lysis = breaking)

Goal: break glucose down to form two pyruvates Who: all life on earth performs glyclolysis Where: the cytoplasm

NOTE: this process does not require O2 and does not yield much energy Before glucose can be converted into ATP, it has to be broken down into two pyruvate molecules (the ionized form of pyruvic acid). This process is known as glycolysis. Glycolysis is a series of chemical reactions during which a fuel molecule ( usually glucose) is degraded and converted into two molecules of a 3-carbon compound called pyruvic acid. The series of chemical reactions involved in glycolysis is shown in figure 6.2 and 6.3 The symbol Pi stands for inorganic phosphate.

Glycolysis takes place in the cytoplasm and can occur without the presence of oxygen and is the primary energy source for most organisms. This process consumes two ATP molecules, and produces four ATP molecules and two NADH2+ molecules. Glycolysis is summarized below:

1. Glucose 6-phosphate is formed when the 6th carbon on the glucose molecule is phosphorylated by an ATP molecule. 2. Glucose 6-phosphate is converted into a 5carbon ring isomer, fructose 6-phosphate. 3. Fructose 6-phosphate is phosphorylated by another ATP to form fructose 1, 6diphosphate. 4. Fructose 1, 6-diphosphate is processed by an enzyme into two glyceraldehyde 3phosphate molecules. 5. Two molecules of glyceraldehyde 3phosphate are oxidized, losing hydrogen atoms and gaining phosphate groups to form 1, 3-diphosphoglycerate. Two molecules of NAD+ are converted into NADH2+ in the process. 6. Two 1,3-diphosphoglycerate molecules phosphorylate ADP (adenine diphosphate) to yield two molecules of 3-phosphoglycerate and two ATPs are produced. 7. The phosphate groups on 3phosphoglycerate move to the 2nd carbon, forming 2-phosphoglycerate. 8. The two 2-phosphoglycerate molecules are dehydrated and forms two high-energy phosphoenolpyruvate molecules. 9. The two phospoenolpyruvate phosphorylates two ADPs and produces two more ATPs and two molecules of pyruvate.

After the glucose molecule has been converted two pyruvate, it is then sent to the Kreb Cycle to be converted into more usable forms of energy.

The First Stage of Glycolysis

PREPARATORY PHASE: Phosphorylation of glucose and its conversion to Glyceraldehyde-3-phosphate This requires two ATP's

The Second Stage of Glycolysis

PAYOFF PHASE ( ENERGY-YIELDING PHASE): Oxidative conversion of glyceraldehydes-3-phosphate to pyruvate and the coupled formation of ATP and NADH

The Individual Reactions of Glycolysis with the Enzymes the Catalyze each reaction

Pathway of glycolysis from glucose to pyruvate. Substrates and products are in blue, enzymes are in green. The two high energy intermediates whose oxidations are coupled to ATP synthesis are shown in red (1,3-bisphosphoglycerate and phosphoenolpyruvate).

Aerobic Respiration

The pyruvate produced in glycolysis undergoes further breakdown through a process called aerobic respiration in most organisms. This process requires oxygen and yields much more energy than glycolysis. Aerobic respiration is divided into two processes: the Krebs cycle, and the Electron Transport Chain, which produces ATP through chemiosmotic phosphorylation. The energy conversion is as follows: C6H12O6 + 6O2 -> 6CO2 + 6H2O + energy (ATP) There are also some important molecules that participate in the process of aerobic respiration. These molecules are the ff:

1. Adenosine Diphoshate ( ADP), and Adenosine Triphosphate (ATP) 2. Nicotinamide Adenine Dinucleotide ( NAD) - is a coenzyme derived

from niacin ( vitamin B5). It acts as hydrogen acceptor during the process of dehydrogenation.

3. Coenzyme A ( abbreviated as COASH) - is a coenzyme derived from

pantothenic acid ( vitamin B). It acts as carrier of acetyl group ( CH3C=O)

4. Flavin Adenine Dinucleotide ( FAD)

- also derived from riboflavin ( vitamin B2) and also acts as hydrogen acceptor.

Oxidation of Pyruvate and the Krebs Cycle (citric acid cycle, TCA cycle)

Goal: take pyruvate and put it into the Krebs cycle, producing NADH and FADH2 Where: the mitochondria There are two steps o The Conversion of Pyruvate to Acetyl CoA o The Krebs Cycle proper The Krebs cycle and the conversion of pyruvate to Acetyl CoA produce 2 ATP's, 8 NADH's, and 2FADH2's per glucose molecule

The pyruvate molecules produced during glycolysis contain a lot of energy in the bonds between their molecules. In order to use that energy, the cell must convert it into the form of ATP. To do so, pyruvate molecules are processed through the Kreb Cycle, also known as the citric acid cycle.

Pyruvate diffuses from the cytoplasm into the mitochondrion. The ff. chemical reactions take place ; 1. one atom of carbon dioxide is released from pyruvate, leaving behind a 2-carbon fragment. 2. This 2-carbon fragment combines with coenzyme A ( COASH) , forming a two-carbon carrier molecule called acetyl coenzyme A or acetyl COA 3. dehydrogenation which involves the release of hydrogen which in turn is accepted by NAD to form NADH.

The Oxidation of Pyruvate to form Acetyl CoA for Entry Into the Krebs Cycle

Krebs Cycle The series of reactions that bring about the breakdown of Acetyl CoA is known as the Krebs Cycle. It is names after its discoverer Hans Krebs. This set of reactions is also known as the Citric acid cycle because the first reaction involves the formation of a 6-carbon compound called citric acid which is a tricarboxylic acid. So it is also known as Tricarboxylic Acid Cycle.

The final steps in the breakdown of glucose, takes place in the Krebs Cycle as shown in Figure 6.3. The ff. chemical changes occur in the each turn of the Krebs Cycle:

1. acetyl 2.

3.

4. 5.

6. 7.

CoA combines with a 4-carbon compound called oxaloacetic acid to form a 6-carbon acid called citric acid. the 6- carbon compound releases CO2 and is converted to a 5carbon compound called ketoglutaric acid. Dehydrogenation also occur during this process wherein hydrogen atoms are removed and accepted by NAD to form NADH ketoglutaric acid in turn gives up CO2 forming a 4-carbon molecule called succinyl coenzyme A. Dehydrogenation also occur in this reaction. Succinyl CoA is converted to a 4-carbon compound called succinic acid with accompanying formation of ATP. Succinic acid in turn undergoes dehydrogenation and is converted to a 4-carbon compound called fumaric acid. This time, FAD acts as the hydrogen acceptor. Fumaric undergoes dehydration ( removal of water) to form a 4-carbon compound called malic acid. Malic acid then undergoes dehydrogenation and is converted back to oxaloacetic acid . NAD acts as the hydrogen acceptor.

Figure 6.4 The Krebs Cycle or Citric Acid Cycle

HYDROGEN TRANSPORT

You will notice that during cellular respiration of glucose, there are several events wherein hydrogen atoms are released ( dehydrogenation) . Where did the hydrogen atoms go?

If you will analyze the series of reactions during cellular respiration, the hydrogen atoms released are accepted by NAD forming NADH. This in turn transfers the hydrogen to the Electron Transport Chain (ETC) where large amount of ATP are generated. The ETC consists of series of hydrogen proton and electron acceptors as illustrated in figure 6.4

Dehydrogenation process is illustrated by the ff. equation:

NAD + 2H ( 2H
+

NADH + H + + 2 e)

The two electrons pass through a series of electron acceptor molecules, each one attracting electrons more strongly than the preceeding acceptor molecule.

So, the electron transport system is a series of chemical reactions into which all of the hydrogens removed from the fuel molecule during glycolysis, oxidation of pyruvate, and the Krebs Cycle, are fed and processed

Electron Transport Chain (ETC)

Goal: to break down NADH and FADH2, pumping H+ into the outer compartment of the mitochondria Where: the mitochondria During this step, the hydrogen ions (or a pair of electrons) are transported from one carrier to another and they are finally used to reduce oxygen to water. During this transfer of electrons, lot of energy is released which is in the form of ATP. ATP is thus an energy rich molecule and can be called the energy currency of the cell.

Fig 6.4 The Electron Transport Chain

The electron transport during aerobic respiration begins with the transfer of electrons from NADH and FADH2 collected on the inner mitochondrial membrane. The electrons move from the high energy level of electron acceptor molecules to the lower energy final acceptor, which is oxygen. Thus, the final product of hydrogen transfer is water.

2 ( 2H + + 2 e ) + O2

2 H2O

As electrons move from one acceptor molecule to another, they gradually give off energy. The cell uses some of this energy to generate ATP. .

THE TOTAL ENERGY YIELD

How much chemical energy does cellular respiration yield from a single molecule of glucose? Recall that glycolysis produces just 6 ATP molecules per glucose. In the absence of oxygen, that is all the energy that a cell can extract from each molecule of glucose.

In the presence of oxygen, everything changes. As shown in table 6.51, the complete breakdown of glucose through cellular respiration, including glycolysis , results in the production of 36 moles of ATP. Table 6.1 Summary of ATP production during the complete breakdown of one molecule of glucose

PROCESS

MOLES PRODUCED

OF

ATP

Glycolysis Pyruvate)

Glucose

to

Oxidation of Pyruvate

Krebs Cycle

24

TOTAL

36

ATP YIELD FROM COMPLETE GLUCOSE METABOLISM Chemical Steps Number of ATP molecules produced -2 4

Glucose Fru-1,6- diphosphate Oxidative phosphorylation 2moles Gly-3-Phosphate 1,3biphosphoglycerate Dephosphorylation 2moles 1,3-biphosphoglycerate pyruvate Oxidative Decarboxylation 2 moles pyruvate acetyl CoA Krebs Cycle TOTAL KREBS CYCLE CHEMICAL STEPS

4 6 24 36

Number of ATP molecules produced 3 3 1 2

Isocitrate Oxalosuccinate - ketoglutarate Succinyl CoA Succinyl CoA Succinate Succinate Fumarate

Malate Oxaloacetate TOTAL

3 12

The oxidation of one acetyl CoA produces 12 ATP molecules and one glucose molecule produce 2 acetyl CoA molecules.

Anaerobic Respiration

Goal: to reduce pyruvate, thus generating NAD+ Where: the cytoplasm Why: in the absence of oxygen, it is the only way to generate NAD+

Alcohol Fermentation - occurs in yeasts in many bacteria o The product of fermentation, alcohol, is toxic to the organism

Lactic Acid Fermentation - occurs in humans and other mammals o The product of Lactic Acid fermentation, lactic acid, is toxic to mammals o This is the "burn" felt when undergoing strenuous activity

The only goal of fermentation reactions is to convert NADH to NAD+ (to use in glycolysis). No energy is gained Note differences anaerobic respiration - 2 ATP's produced (from glycolysis), aerobic respiration - 36 ATP's produced (from glycolysis, Krebs cycle, and Oxidative Phosphorylation)

Thus, the evolution of an oxygen-rich atmosphere, which facilitated the evolution of aerobic respiration, was crucial in the diversification of life