Lesson Objective: Describe aerobic and anaerobic respiration and the requirement for such conditions.

5.1 TYPES OF RESPIRATION

There are 2 types of cell respiration, namely a) Aerobic Respiration Which requires O2, summarized by the following equation: C6H12O6 + 6O2 6CO2 + 6H2O + Energy

b) Anaerobic Respiration which does not require O2; the equations of the reaction are different in plants & animals.
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 In Plants C6H12O6

2C2H2OH + 2CO2 + 2ATP (ethanol)

In Animals C6H1206

2CH3CH(OH)COOH + 2ATP (lactic acid)

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Adenosine triphosphate (ATP)

Adenosine triphosphate (ATP) is a nucleotide molecule. It consists of a base adenine, a pentose sugar ribose, combined with three phosphate groups.

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 When ATP is hydrolysed, ADP & phosphate (Pi) are produces & energy is released.

ATP + H2O

ATPase

Hydrolyse

ADP + Pi +30.6kJ
Per mol ATP

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DEFINITION
Cellular respiration refers to a series of biochemical reactions that take place in the cells. This process involves the breakdown of organic molecules to liberate energy (ATP).

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Lesson Objective: Describe the importance of aerobic respiration during oxidation of glucose which involve glycolysis, Krebs cycle & electron transport chain.

5.2:Aerobic Respiration
There are 4 main stages in aerobic respiration; i) Glycolysis ii) Link Reaction iii) The Krebs Cycle iv) Electron Transport Chain In eukaryotes, glycolysis occurs in the cytoplasm The other processes take place in the mitochondria.

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Lesson Objective: Outline glycolysis from glucose to pyruvate with the yield of ATP & reduced NAD+.

GLYCOLYSIS
Glycolysis is the breakdown of the glucose (6C) molecule in a number of enzymecontrolled steps into 2 molecules of pyruvate (3C). The process takes place in the cytoplasm of cells & does not require O2. There is a net production of 2 ATP molecules per molecule of glucose.
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Lesson Objective: Show the steps where the energy, ATP and NADH are produced.

STAGE 1

STAGE 2

STAGE 3 (X 2)

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An outline of the main stages of glycolysis

Stage 1

: Phosphorylation of Glucose

The glucose molecule is phosphorylated, receives a high energy phosphate from ATP to increase its energy level to become glucose-6phosphate. ATP Glucose ADP + Pi glucose-6-phosphate
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2. Glucose-6-phosphate is rearranged to become the isomer fructose-6-phosphate. 3. The frutose-6-phosphate is further activated by the addition of another phosphate group from ATP.
ATP ADP + Pi

Fructose-6-phosphate

fructose-1,6diphosphate

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Stage 2 The fructose-1,6-diphosphate produced is split (lysis) into gluceraldehyde-3-phosphate & its isomer dihydroxyacetone phosphate. Fructose-1,6-diphosphate G3P + DHAP
Isomerase

:Breakdown of fructose diphosphate

G3P DHAP (3C) (3C) DHAP rearranges into another molecule of G3P (2 molecules of G3P produced go through identical reaction).

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Stage 3

:Oxidation of G3P

G3P is oxidised, hydrogen atoms are removed, NAD+ is reduced to become NADH. An inorganic phosphate (Pi) attached to increase the energy of glycerate-1,3diphosphate. One phosphate from each glycerate-1,3diphosphate is transferred to ADP to form ATP.

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 The 3-phosphoglycerate is rearranged to form 2phosphoglycerate. Removal of water produces phosphoenolpyruvate (PEP). The 2nd phosphate is transferred to ADP form ATP. Phosphoenol pyruvate is converted to pyruvate (pyruvic acid). A summary of ATP & NADH production during glycolysis of a glucose molecule is

2 molecules of ATP (4-2 molecule is that were used in stage 1. 2 molecules of NADH.
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Lesson Objective: Out line the link reaction to explain the conversion of pyruvate to acetyl coenzyme A (2C), before entering the Krebs cycle.

2. LINK REACTION
Aerobic respiration takes place when O2 is available. Pyruvate easily enters the matrix of the mitochondria. Pyruvate (3C) formed at the end of glycolysis is decarboxylated (removal of CO2) & is oxidised (the removal of hydrogen atoms) to form 2carbon acetate (2C). The acetate combines with coenzyme A (Co A) to form 2-carbon acetyl coenzyme A (Acetyl-Co A) which then enters into the Krebs Cycle.

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Link reaction

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 The process is called oxidative decarboxylation or the link reaction as it links glycolysis to Krebs Cycle. There are 2 acetyl Co A molecules formed since one glucose molecule produces 2 pyruvate molecules. (The link reaction is sometimes included as part of the Krebs Cycle).

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Lesson Objective: Outline the Krebs cycle occuring in the mitochondrion, explaining that citrate is reconverted to oxaloacetate in a series of reactions.

3. KREBS CYCLE
Acetyl Co A (2C) combines with oxaloacetate (4C) in a condensation reaction to form citrate (6C). A coenzyme (Co A) is released. Citrate rearranges by the removal of a water molecule & the addition of water to form its isomer isocitrate (6C). Isocitrate is oxidised, yeilding a pair of electrons that reduce a molecule of NAD+ to NADH.
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Lesson Objective: State the steps where the energy and ATP, NADH and FADH2 are produced.

Then the oxidised intermediate is decarboxylated, yielding a five carbon molecule called -ketoglutarate (5C). Second oxidative-decarboxylation of ketoglutarate takes place. This produces succinyl coenzyme A (4C), C02 & NADH. Substrate level phosphorylation take place. Succinyl Co A is converted to succinate (4C). The energy released is used for phosphorylation of GDP forming GTP. GTP transfer its phosphate group to ADP forming ATP.
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 Succinate is oxidised to fumarate (4C), 2H atoms are transferred to FAD to form FADH2. Fumarate becomes hydrated by addition of water is converted to malate (4C). Malate is oxidised regenerating oxaloacetate (4C) & NAD+ is reduced to NADH. Oxaloacetate can be used to combine with acetyl Co A & the cycle is repeated.
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Co A

(4C)

(6C) (6C)

(4C)

KREB CYCLE
(5C)

(4C)

(4C)

(4C)

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Lesson Objective: Explain the processes involved in decarboxylation and dehydrogenation and describe the role of NAD+ and FAD.

In this cyclic process, decarboxylation takes place twice, dehydrogenation takes place 4 times & formation of GTP from ADP & phosphate once. During the dehydrogenation process, 3 times NAD+ & one time FAD are used as H acceptors forming NADH + H+ & FADH2 respectively. From one Krebs Cycle; 3 NADH, one FADH2 & one GTP are produced. One glucose molecule produces 2 molecules of acetyl coenzyme. The cycle is turned twice for each glucose molecule broken down.
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Lesson Objective: Describe oxidative phosphorylation occurring in the mitochondrion including the role of oxygen.

4. THE ELECTRON TRANSPORT CHAIN

The electron transport system is a chain of electron acceptor embedded in the inner membrane of the mitochondrion. High energy electron removed from respiratory intermediates are carried by NADH & FADH2 to inner mitochondrial membrane. The folding of the inner membrane to form cristae increases its surface area, providing space for thousands of copies of the chain in each mitochondrion. During electron transport along the chain, electron carriers alternate between reduced and oxidized states as they accept and donate electrons.

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 Complex I: NADH Dehydrogenase ( Flavoprotein). Complex I is responsible for removing two electrons from NADH and transferring them to the electron carrier, ubiquinone (Coenzyme Q). NADH dehydrogenase also moves four protons from the mitochondrial matrix to the intermembrane space, beginning the production of a proton gradient.
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 Complex II: Succinate Dehydrogenase: Complex II removes electrons from succinate and transfers them to ubiquinone via FAD. Succinate dehydrogenase does not contribute to the proton gradient. Complex III: Cytochrome b-c Complex: Complex III removes two electrons form ubiquinone and transfers them to two molecules of the electron carrier, cytochrome c. The cytochrome b-c complex also moves four protons across the inner mitochondrial membrane, further contributing to the proton gradient.
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 Complex IV: Cytochrome Oxidase complex: Complex IV removes two electrons from the two molecules of cytochrome c and transfers them to molecular oxygen (The final electron acceptor) which combined with H+ ions to form water. O2 + 2e- + 2H+ H2 O

Cytochrome c oxidase also moves two electrons across the inner mitochondrial membrane, adding to the proton gradient.
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 As they pass along the electron transport chain, they lose much of their energy & the energy released is used to synthesise ATP. Another source of electron for electron transport chain is FADH2, the other reduced product of the Krebs cycle. FADH2 adds its electron to the electron transport chain at a lower energy level than NADH does (at ubiquinone).
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 Consequently, the electron transport chain provides less energy for ATP synthesis when the electron donor is FADH2. 3 ATP molecules are produced for every NADH that enters the electron transport chain.

2 ATP molecules for every FADH2 that enters the electron transport chain.
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Lesson Objective: Explain the chemiosmosis theory.

Chemiosmosis Hyphothesis
Electrons released by the oxidation of substrate in the matrix flows down the electron transport chain. The energy released by the electron transport chain is used to pump hydrogen ions (H+) from the matrix into the intermembrane space. This builds up a transmembrane electro-chemical protons (H+) gradient.
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 The inner mitochondrial membrane is permeable to hydrogen ions. The gradient forces hydrogen ions to diffuse through the ATP synthetase (ATP synthase) complex down its electrochemical gradient. Their potential energy is used to synthesise ATP from ADP & Pi. The process is called chemiosmosis.
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Energy yield from the complete oxidation of glucose by aerobic respiration

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Mitochondrial shuttle system

The inner motochondrial membrane is not permeable to NADH. Therefore the NADH molecules produced in the cytosol during glycolysis cannot diffuse into the mitochondria to transfer their electrons to the electron transport chain. Unlike ATP & ADP, NADH does not have a carrier protein to transport it across the membrane.

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 In liver, kidney, & heart cells, a special shuttle system transfers the electrons from NADH through the inner mitochondrial membrane to the NAD+ molecule in the matrix. These electrons are transferred to the electron transport chain in the inner mitochondrial membrane, and up to three molecules of ATP are produced per pair of electron.
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 In skeletal muscle, brain & some other types of cells, another type of shuttle operates. Because this shuttle requires more energy than the shuttle in liver, kidney & heart cell, the electron are at a lower energy level when they enter the electron transport chain. They are accepted by ubiquinone rather than by NAD+ & so generate a maximum of 2 ATP molecules per pair of electrons. This is why the number of ATPs produced by aerobic respiration of 1 molecule of glucose in skeletal muscle cells is 36 rather than 38.

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5.3: Anaerobic Respiration : Fermentation & Application

In the absence of O2, anaerobic respiration occurs, for e.g: a) in the several types of fungi, bacteria & earthworms living in muddy & O2 deficient conditions. b) in skeletal muscles of vertebrates that are contracting actively.
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 Organisms that obtain energy from the breakdown of sugar through anaerobic respiration are known as anaerobes. During anaerobic respiration, glucose only breaks down into pyruvic acid when undergoing glycolysis. The pyruvic acid formed is not converted into acetyl Co A for entry into Krebs cycle as in aerobic respiration. Instead, the pyruvic acid is converted into ethanol or lactic acid.

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Lesson Objective: Explain what is meant by fermentation Describe alcohol and lactate fermentation

Fermentation
Anaerobic mechanisms of energy production which do not involved the respiratory chain or cytochromes are called fermentation Fermentation coverts glucose into lactic acid (lactate) in animal cells (Lactate fermentation) Fermentation converts glucose into ethanol & CO2 in plant cells, fungi cells (eg; yeast ) & bacteria (Alcohol fermentation).

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Lactate (lactic acid ) Fermentation

Occurs in certain fungi, bacteria & during strenuous activity in muscle cells of humans & other complex animal. In this alternative pathway, NADH produced during glycolysis transfers hydrogen atoms to pyruvate, reducing it to form lactate & NAD+ is regenerated.

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 If the amount of O2 delivered to muscle cells is insufficient to support aerobic respiration, the cells shift briefly to lactate fermentation. Lactate accumulation in the muscle causes fatigue & muscle cramps. The O2 that is required to break down the lactate is known as the oxygen debt & is repaid by deep & rapid breathing at the end of strenuous activity.

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 The lactate formed is removed to other tissues and dealt with by one of two mechanisms :
it is converted back to pyruvate The pyruvate then proceeds to be further oxidised, finally producing a large amount of ATP. it is converted back to glucose in the liver

The process of conversion of lactate to glucose is called gluconeogenesis, uses some of the reactions of glycolysis (but in the reverse direction) and some reactions unique to this pathway to re-synthesise glucose.
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Lesson Objective:

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Alcohol Fermentation
E.g: Yeast carried out alcohol fermentation when deprived of O2. They have enzymes that decarboxylate pyruvate, releasing CO2 & forming a twocarbon compound called acetaldehyde. NADH produced during glycolysis transfers hydrogen atoms to acetaldehyde, reducing it to form ethyl alcohol (ethanol) & NAD+ regenerated to be reused.
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 Both alcohol fermentation & lactate fermentation are highly inefficient because the fuel is only partially oxidised. This is because a considerable quantity of energy still remains trapped in the ethanol or lactic acid molecules. A net profit of only 2ATPs is produced by the fermentation of one molecules of glucose, compared with up to 36-38ATPs when O2 is available.
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Lesson Objective: Discuss the importance of fermentation in industry.

Importance of Fermentation in Industry. Yeast

a) In the process of baking cakes & bread In this process, the flour dough is mixed with yeast. The CO2 gas produced from the fermentation process causes the dough to rise & give soft cake or bread texture when it is baked in the oven.

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b) In the process of beer & wine manufacture During the manufacture of beer, the enzyme (diastase) in the malt, rice or corn convert the starch in the cereal into maltose. The yeast mixture is then added to allow fermentation to take place. During fermentation, the enzyme maltase converts maltose into glucose. Glucose is then converted by the enzyme zymase into ethanol & CO2. Wine is made by the fermentation of yeast on grape or other fruit sugars.

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c) In the process of making Cheese & Yoghurt (Dairy Industry). Cheese is made from milk. Several type of bacteria & fungi are used to form different type of cheese through lactate fermentation.
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Yoghurt is made of concentrated milk. Lactose in the milk is fermented to lactic acid by lactic acid bacteria, e.g; Lactobacillus bulgaricus. Lactic acid gives the yoghurt its flavour (sour taste).

Lactobacillus

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Local fermented foods

Tempe is one of the most popular fermented foods in Indonesia, Malaysia and Singapore. It is traditionally prepared with soy beans or a certain variety of peanut fermented with mold, Rhizopus spp. The cultured soybeans or nuts are bound together by a thick white mycelium of new moldgrowth, to form a cake.
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 Another examples of local fermented foods are tapai, dadih, budu and cincalok. Ragi (yeast cake) is used by crushing it, and then mixing the powder with cooked, cooled ingredients such as glutinous rice (for tapai making). The mixture is fermented for a particular length of time, depending on the product being prepared.
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Lesson Objective: Briefly describe how lipid is oxidised through -oxidation.

Alternative sources of energy

The Oxidation of Fatty Acid Fats (or lipids), like triglycerides, are also metabolized to produce energy. Triglycerides Glycerol + Fatty Acids. Glycerol Glyceraldehyde Pyruvic Acid Acetyl CoA Kreb's Cycle Each fatty acid molecule is oxidized by a process called -oxidation. This reaction not only produces lots of Acetyl CoA (or acetate), but lots of hydrogen. The Acetyl CoA goes through the Kreb's Cycle, while the hydrogen go through Oxidative Phosphorylation. The process takes place inside the mitochondrial matrix.
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Lesson Objective: Briefly describe how protein is oxidised through transamination and deamination.

Each gram of lipid contains 9 kcal (38kJ), more than twice as much energy as 1 g of glucose or amino acid, which have about 4kcal (17kJ) per gram. The oxidation of amino acid When carbohydrate & lipid reserve have been exhausted, amino acid derived from protein digestion are also can used as fuel molecules. Amino acids are transformed into one of the metabolic intermediates that are fed into glycolysis or the Krebs cycle.
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 Proteins first hydrolysed into their monomers amino acids & deaminated (their amino group {-NH2} are removed). The amino group is converted to urea & excreted. The carbon chain is metabolised & eventually is used as a reactant in one of the steps of respiration. The amino acid alanine, for eg., undergoes deamination to become pyruvate, the amino acid glutamate is converted to ketoglutarate & the amino acid aspartate yields oxaloacetate. Pyruvate enters aerobic respiration as the end product of glycolysis, & -ketoglutarate & oxaloacetate both enter aerobic respiration as intermediates in the Krebs Cycle.

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