AP BIOLOGY · UNIT 3 · CELLULAR ENERGETICS

How a cell turns food into ATP

Every cell in your body is pumping, building and moving right now, and every one of those jobs is paid for in ATP. Respiration remakes it, releasing energy from glucose and fat a step at a time. This is how it works with oxygen and without, where the protons come in, and how to measure it in a tube of peas.

AP Biology 3.53 lessons10 chaptersdry lab

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ribosomesDNAmitochondrionglucoseoxygencarbondioxidewaterpumpingbuildingmovingATPATPATP ribosomesDNAmitochondrionglucoseoxygencarbon dioxidewaterpumpingbuildingmovingATPATPATP
Glucose and oxygen in, carbon dioxide and water out: the mitochondrion makes ATP, and every job in the cell spends it.

01Learn

Ten chapters. Read one, play its figure, and make each prediction before you open the answer. The film after chapter 2 follows one glucose through a muscle cell; then try keeping a muscle cell running yourself.

  1. 1Fuel in, ATP out
  2. 2With oxygen and without
  3. 3A rate you cannot see
  4. 4Taking hydrogen away
  5. 5Two ATP in, four out
  6. 6Giving NAD+ back
  7. 7The carbon leaves as carbon dioxide
  8. 8Down the chain
  9. 9Protons through the turbine
  10. 10Fat or sugar?

3.5.A Respiration releases the energy in food as ATP

Fuel in, ATP out

Cell respiration is how a cell makes ATP, using energy released from carbon compounds. It happens in every living cell, all the time, because ATP is used all the time and only a little is kept (ATP and coupling, in the enzymes guide). That includes plants: a leaf cell respires by day and by night.

The fuels are called respiratory substrates. The main two are glucose and fatty acids, but a cell can use a wide range of carbon compounds, amino acids among them when food is short. Glycogen in liver and muscle, and starch in a plant, are broken down to glucose first.

Respiration releases the energy a step at a time, each step helped by its own enzyme at body temperature. Burning glucose is combustion. It starts with glucose and oxygen and ends with carbon dioxide and water, as respiration does, and it releases the same total energy. But combustion releases it all at once, as heat and light, and makes no ATP. One is controlled and useful to a cell, the other is not.

Respiration is not gas exchange. Respiration is chemistry inside cells. Gas exchange is oxygen and carbon dioxide diffusing across a surface, in the lungs, the gills or a leaf, with no reaction and no energy released. The carbon dioxide you breathe out was made in the mitochondria of your cells, and it only leaves through your lungs. Breathing moves the air; that is not respiration either.

The map under this chapter is the whole of respiration on one page: where each stage happens, what goes in, what comes out and where the ATP is made. Come back to it whenever a stage is new.

Chapters 4 to 10 open up each stage, and each starts with a small copy of the map, marking where you are.

Watch out “Respiration” in everyday English means breathing. In biology it is the chemistry that makes ATP in every cell. Breathing moves air; gas exchange moves gases across a surface; neither releases energy.
Watch out Respiration does not “make energy”. Energy is released from the fuel and transferred to ATP; none is created.

A leaf in bright light takes in carbon dioxide and gives out oxygen. Is it respiring?

Burning a fuelglucoseoxygencarbon dioxidewaterheat and lightfood on amounted needleReleased all at once, as heat and light.No ATP is made.Cell respirationa cellglucoseoxygencarbon dioxidewaterATPATPATPan enzymefor each stepenergy, step by stepReleased a step at a time, each step its own enzyme:ATP is made, and some heat.Gas exchangeair in anair sacbloodred bloodcellsoxygencarbon dioxidecarbon dioxidefrom the body's cellsDiffusion across a surface: no reaction,and no energy released. Burning a fuelglucoseoxygencarbondioxidewaterheat and lightfood ona needleReleased all at once,as heat and light.No ATP is made.Cell respirationa cellglucoseoxygencarbondioxidewaterATPATPan enzymeeach stepenergy, stepby stepReleased a step at a time,each step its own enzyme:ATP is made, and some heat.Gas exchangeair in anair sacblood:red blood cellsoxygencarbon dioxidecarbon dioxide fromthe body's cellsDiffusion across a surface:no reaction,and no energy released.

Burning: glucose and oxygen in, carbon dioxide and water out. The energy is released all at once, as heat and light, and no ATP is made.Cell respiration: glucose and oxygen in, carbon dioxide and water out. The energy is released a step at a time, each step with its own enzyme, and ATP is made.Gas exchange: oxygen diffuses from the air in an air sac into the blood, and carbon dioxide the other way. No reaction, and no energy released.

Three different things. What goes in, what comes out, and what is released?
ribosomesDNAcytoplasmmitochondrionits electronsare passed in2 pyruvatematrixinner membranefolded into cristae2 acetyl CoAglucose2 ATPnet: 2 used, 4 made2 NADHglycolysisno more ATPlactatein yeast: ethanoland carbon dioxideno oxygen2 carbon dioxide2 NADHpyruvate oxidation4 carbon dioxide2 ATP6 NADH2 FADH2Krebscyclepumps protons into theintermembrane spaceoxygenwaterelectron transportchainprotons flow backabout 26 to 28 ATPATP synthaseintermembrane spaceabout 30 to 32 ATPin all, from one glucoseribosomesDNAcytoplasmmitochondrion2 pyruvatefolded innermembraneglucose2 ATPglycolysisno more ATPlactatein yeast: ethanoland carbon dioxideno oxygen6 oxygen6 waterabout 28 to 30 more ATP6 carbon dioxidewith oxygenabout 30 to 32 ATPin all, from one glucose
Where each stage happens, what goes in and what comes out. Tap a stage to read what happens there.

3.5.A With oxygen and without

With oxygen and without

cytoplasmmitochondrionglycolysislactateno oxygencarbon dioxidewaterabout 28 to 30 more ATPwith oxygenyou are here

Both kinds of respiration start the same way, in the cytosol. A glucose molecule is split into two molecules of pyruvate in a process called glycolysis, which releases enough energy to make a small amount of ATP. After that the routes divide, according to whether there is oxygen, and they differ most in their yield of ATP for each glucose.

Aerobic respiration goes on with oxygen. The pyruvate goes into a mitochondrion and is broken down completely, to carbon dioxide and water, and a large amount of ATP is made. The word equation is glucose + oxygen → carbon dioxide + water. It needs mitochondria and oxygen, and it can use fatty acids and other compounds as well as glucose.

Anaerobic respiration in humans stops early. When there is not enough oxygen, the pyruvate stays in the cytosol and is turned into lactate. The word equation is glucose → lactate. It needs no mitochondria, no oxygen, and makes no carbon dioxide, but only glucose can be used: fatty acids cannot be respired without oxygen.

When does a human cell respire anaerobically? When muscles work so hard that the blood cannot bring oxygen fast enough: a short sprint, a heavy lift. Anaerobic respiration gives ATP at once, without waiting for oxygen, but only for a short time. Muscles do not switch from one route to the other. Aerobic respiration goes on, and anaerobic respiration adds to it.

Why make lactate at all, rather than stopping at pyruvate? Chapter 6 explains.

A sprinter’s leg muscles run short of oxygen. Per glucose, how much ATP does anaerobic respiration make compared with aerobic?

Remember it Anaerobic in humans is LOW

  • LLactatethe waste product
  • OOnly 2 ATPper glucose
  • WWithout oxygenin the cytosol
Watch out Glycolysis happens in the cytosol in both routes; only what follows it happens in the mitochondria.
Watch out The waste product in human muscle is lactate, not carbon dioxide. People often call it lactic acid. Lactate is not what makes muscles ache the next day.
capillarycytosol of a muscle cellmitochondrionglucoseglycolysis2 ATP2 pyruvatewith oxygenoxygencarbon dioxidewaterATP made per glucoseabout 30 to 32one tile, one ATP (the last two dashed)2 lactatelactate into the bloodlittle oxygen: the mitochondrion can do littleATP made per glucose2, from glycolysisone tile, one ATPglucose + oxygen → carbon dioxide + waterglucose → lactate capillarymuscle cellglucoseglycolysis2 ATP2 pyruvatemitochondrionwithoxygenoxygencarbon dioxidewaterATP made per glucoseabout 30 to 32one tile, one ATP2 lactatelactate into the bloodlittle oxygen:little done hereATP made per glucose2, from glycolysisone tile, one ATPglucose + oxygen →carbon dioxide + waterglucose → lactate

Plenty of oxygen: glucose is split into two pyruvate in the cytosol, and the mitochondrion breaks them down to carbon dioxide and water. About 30 to 32 ATP per glucose.Not enough oxygen: the pyruvate becomes lactate in the cytosol and goes out into the blood. 2 ATP per glucose, from glycolysis.

One glucose in a muscle cell. Switch the oxygen supply and follow the pyruvate.
Aerobic and anaerobic respiration in humans
AerobicAnaerobic
Substratesglucose, fatty acids and other carbon compoundsglucose (carbohydrate) only
Oxygen needed?yesno
ATP per glucoseabout 30 to 322
Waste productscarbon dioxide and waterlactate
Where in the cellglycolysis in the cytosol, the rest in the mitochondriathe cytosol
Mitochondria needed?yesno
Word equationglucose + oxygen → carbon dioxide + waterglucose → lactate
Watch one glucose give up its energy in a muscle cell. Then turn the oxygen off.
oxygenwithout oxygenATPa phosphateribosomeglycogenmyofibrilglucosemitochondrionsodiumionsglucosetransporterscapillarylactate42 ATP + lactate33ADP 7about 30 ATPsodium-potassiumpumpglycolysisflow cutlactate02 ATP + lactate0ADP 40about 30 ATPsodium-potassiumpumpcapillarylactate222 ATP + lactate14ADP 26about 30 ATPsodium-potassiumpumpflow cutNADHNADHNADHNADHNADHNADHmake lactatefree NAD+: 0 of 69ADP 31no oxygen: idlesodium-potassiumpump
  1. With oxygen, glycolysis splits each glucose in the cytosol and its two pyruvate go into the mitochondrion: about 30 ATP in all. Without oxygen, glycolysis alone: 2 ATP and lactate.
  2. Cut the supply: how long does a working fiber's ATP last?
  3. The sprint: the oxygen cannot keep up. Where does the extra ATP come from?
  4. No oxygen and no lactate made: watch the NAD+.

3.5.A Measuring the rate of respiration

A rate you cannot see

Respiration happens inside cells, so you cannot watch it. You measure it from what goes in or comes out: oxygen taken up, carbon dioxide given out, or fuel used. The rate of respiration is the amount changed divided by the time taken, and for living material it is given per gram per minute as well, so that samples of different mass can be compared.

A class can do it three ways.

  • Oxygen uptake in a respirometer: germinating seeds in a sealed tube, with potassium hydroxide to absorb the carbon dioxide they give out, and a drop of colored liquid in a capillary whose movement shows the change in gas volume.
  • Carbon dioxide output: a gas sensor or data logger in a sealed chamber, or hydrogencarbonate indicator changing color. With yeast, the rise of a dough or the gas collected over a sugar solution works too (the dry lab's dough mode).
  • Fuel used: the fall in the glucose in a yeast culture, measured with test strips.

The capillary is a long thin cylinder, so the volume of gas is πr² × the distance the drop moved, where r is half the bore. A bore of 1 mm holds 0.785 mm³ for every mm.

A second tube, the same in every way but with glass beads in place of the seeds, is the control. Anything that moves its drop, such as the gas warming or cooling or the air pressure changing, is not respiration, so its movement is taken off the seeds’.

What changes the rate? Temperature: respiration is enzymes at work, so it speeds up as it warms until they begin to denature (topic 3.2). How far germination has gone: dry seeds hardly respire, and soaked ones respire more each day. The mass of the seeds, and the kind of seed. With yeast: the sugar it is given, the temperature, and how much oxygen there is. The seeds are kept in the dark so that they cannot photosynthesize and hide the gas changes.

In an experiment on any of these, the one thing you change on purpose is the independent variable (the temperature, say), the rate you measure is the dependent variable, and everything else you hold the same is a controlled variable: the mass and age of the seeds, the volume of the tube, the time. You may also be asked to work out a rate from someone else’s numbers: the steps are the same, as in the second figure.

Watch out The drop does not move because the seeds give out carbon dioxide: that is removed. It moves because of what the seeds take up.
Watch out Germinating seeds are not “breathing”. They have no lungs; their cells respire, and the gases cross their surfaces by diffusion.

In the respirometer the seeds respire and the potassium hydroxide absorbs their carbon dioxide. Which way does the drop move?

water baththermometergerminatingseedsglass beads:the controlwire gauzepotassiumhydroxidebung andscrew clipcapillary and colored drop0100200mmscale thermometerwater bathgerminatingseedsglass beads:the controlwire gauzepotassiumhydroxidebung andscrew clipcapillary and colored drop0100200scale, mm

Tap a part of the respirometer.

Try it on the bench The dry lab has this respirometer set up and ready: run it, take readings and work out the rate.

germinating seeds, 3.0 g010203040mm24 mmdrop at 0 minat 4 minglass beads: the control010203040mm2 mmcorrected distance = 24 − 2 = 22 mmvolume = π r2 × distance = 3.14 × 0.52 × 22 = 17.3 mm3rate = 17.3 mm3 ÷ 4 min ÷ 3.0 g = 1.44 mm3 g−1 min−1a bore of 1 mm, so r = 0.5 mm germinating seeds, 3.0 g010203040mm24 mmglass beads: the control010203040mm2 mmdrop at 0 minat 4 mincorrected distance = 24 − 2 = 22 mmvolume = π r2 × distance= 3.14 × 0.52 × 22 = 17.3 mm3rate = 17.3 mm3 ÷ 4 min ÷ 3.0 g= 1.44 mm3 g−1 min−1a bore of 1 mm, so r = 0.5 mm
  1. Read both drops at the start and the end.
  2. Take off the control’s movement.
  3. Turn distance into volume with the bore.
  4. Divide by time and by mass.

3.5.A Redox and the hydrogen carrier NAD

Taking hydrogen away

In respiration, energy is released from fuel molecules by taking hydrogen off them. To see why, you need two words. Oxidation is the loss of electrons, and reduction is the gain of electrons. They always happen together: what one molecule loses, another gains. A reaction that does both is a redox reaction.

A hydrogen atom is a proton and an electron. So when hydrogen is removed from a molecule, which is called dehydrogenation, it loses electrons too, and has been oxidized. The enzymes that do it are dehydrogenases.

The hydrogen has to go somewhere. It is accepted by NAD+, a molecule built from nucleotides, like ATP. When NAD+ accepts hydrogen it is reduced, and is then called NADH. NADH carries the hydrogen, and the energy that comes with it, to where it is used (the electron transport chain, chapters 8 and 9), and is turned back into NAD+ there. So NAD+ is a hydrogen carrier.

A cell has only a little NAD+, so it must be unloaded and used again and again, as ATP is. Every stage of respiration that follows either loads NAD+ or depends on NAD+ being given back (chapter 6).

Why does it matter? A large part of the energy in a fuel is in its hydrogen and the electrons that go with it. Respiration is, step by step, taking hydrogen off the fuel and passing it to oxygen: that is the role of hydrogen, and of oxygen, in the release of energy.

Remember it Oxidation and reduction

  • OilOxidation Is Lossof electrons
  • RigReduction Is Gainof electrons
Watch out NADH carries hydrogen, protons and electrons together, and the energy that comes with them. It is not ATP and it is not “energy” itself.

A dehydrogenase takes two hydrogens off a molecule of fuel. What has happened to that molecule?

hydrogen atomprotonelectronsubstrate(fuel)active sitedehydrogenase (an enzyme)NAD:a hydrogen carrierNADNADtwo hydrogens:protons and electronsNADoxidized: lost electronsNADH:gained electronsNADNADH,to the chainNADNAD comes backfor more hydrogen atomprotonelectronsubstrate(fuel)dehydrogenase (an enzyme)its active site holds the fuelNAD:a hydrogencarrierNADNADtwo hydrogens:protons and electronsNADoxidized:lost electronsNADH:gained electronsNADNADH,to the chainNADbackfor more
  1. A substrate, an enzyme, and NAD+.
  2. Hydrogen is removed: dehydrogenation. Each hydrogen is a proton and an electron.
  3. The substrate is oxidized; NAD+ is reduced. A redox reaction.
  4. NADH carries the hydrogen away, and NAD+ comes back for more.

3.5.A Glycolysis

Two ATP in, four out

cytoplasmmitochondrionNADH and FADH2carried to the chainglycolysislactateno oxygenpyruvate oxidationKrebs cycleelectrontransportchainyou are herecytoplasmmitochondrionglycolysislactateno oxygencarbon dioxidewaterabout 28 to 30 more ATPwith oxygenyou are here

Glycolysis turns one glucose into two pyruvate, in the cytosol, in about ten steps. Each step has its own enzyme, and the product of one step is the substrate of the next: it is a metabolic pathway (topic 3.3). The course asks for the four stages, not for the names of the molecules in between.

  • Phosphorylation: two ATP each give a phosphate group to the glucose. That costs 2 ATP, but a sugar carrying two charged phosphates is made reactive and ready to split.
  • Lysis: the 6-carbon sugar with its two phosphates splits into two 3-carbon sugars, each with one phosphate.
  • Oxidation: hydrogen is taken off each 3-carbon sugar and accepted by NAD+, so two NADH are made for the glucose. The energy released lets each 3-carbon sugar take on a second phosphate, a free one from the cytosol, not one from ATP.
  • ATP formation: each 3-carbon sugar passes its two phosphates to ADP, making ATP directly, and ends as pyruvate.

Glycolysis spends 2 ATP at the start. How many does it make by the end, from one glucose?

Remember it The four stages of glycolysis, in order

  • PenguinsPhosphorylation2 ATP spent
  • LoveLysis6 carbons split into 3 and 3
  • OrangeOxidationhydrogen to NAD+
  • AnchoviesATP formation4 ATP made
Watch out Glycolysis does not need oxygen, and it does not happen in the mitochondrion.
In the cytosolcarbonphosphatehydrogenan enzyme: each step has its own1 Phosphorylation2 Lysis3 Oxidation4 ATP formationglucose6 carbons2 ATP2 ADP6C sugar withtwo phosphates3C sugarphosphateNADNADHNADNADHfreephosphate3C sugar withtwo phosphates2 ADP2 ATP2 ADP2 ATPpyruvate3 carbons12345667891078910ATP spent2NADH made2ATP made4net gain2 ATP In the cytosolcarbonphosphatehydrogenan enzyme: each step has its ownglucose6 carbons1 Phosphorylation2 ATP2 ADP6C sugar withtwo phosphates2 Lysistwo 3C sugarphosphates3 Oxidation2 NAD2 NADHfreephosphatetwo 3C sugars,2 phosphates each4 ATP formation4 ADP4 ATPtwo pyruvate3 carbons each12345678910ATP spent2NADH2ATP made4net gain2
  1. One glucose, in the cytosol.
  2. Phosphorylation: 2 ATP spent, the sugar made reactive.
  3. Lysis: split into two 3-carbon sugars.
  4. Oxidation: hydrogen to NAD+, making 2 NADH.
  5. ATP formation: 4 ATP made, so 2 gained.
  6. Ten steps, each with its own enzyme: two pyruvate, on to the link reaction.

3.5.A Fermentation: giving NAD back

Giving NAD+ back

cytoplasmmitochondrionNADH and FADH2carried to the chainglycolysislactateno oxygenpyruvate oxidationKrebs cycleelectrontransportchainyou are herecytoplasmmitochondrionglycolysislactateno oxygencarbon dioxidewaterabout 28 to 30 more ATPwith oxygenyou are here

Glycolysis needs NAD+. Every glucose that goes through it turns two NAD+ into NADH. A cell has only a little NAD+, so if NADH is not turned back into NAD+, glycolysis stops very quickly. Giving NAD+ back is called the regeneration of NAD+.

With oxygen, NADH is unloaded at the electron transport chain in the mitochondrion (chapters 8 and 9). Without oxygen it cannot be, so a cell needs another way to give NAD+ back.

In human muscle, pyruvate takes the hydrogen from NADH and becomes lactate. NAD+ is given back, so glycolysis can go on, with a net yield of 2 ATP per glucose. Making lactate releases no extra energy: its whole point is to give NAD+ back.

Yeast follow the same pathway as far as pyruvate. Without oxygen, each pyruvate loses a carbon as carbon dioxide, and what is left takes the hydrogen from NADH to become ethanol. NAD+ is given back, and the yield is again 2 ATP per glucose. Only the last step differs from humans, and so the final products do.

Bakers and brewers use what yeast make. In baking, yeast respire the sugars in the dough and the carbon dioxide forms bubbles that stretch it, so the dough rises; the ethanol evaporates in the oven. In brewing, yeast in a sugary liquid made from barley or grapes, with little air, make ethanol, which is kept, while the carbon dioxide escapes or is kept in for fizz. Early in a dough or a brew there is oxygen and the yeast respire aerobically, so carbon dioxide is given off from the start; as the oxygen runs out, anaerobic respiration takes over. Anaerobic respiration in yeast is often called fermentation.

Without oxygen, why does a muscle cell turn pyruvate into lactate instead of leaving it as pyruvate?

Watch out Yeast without oxygen make ethanol and carbon dioxide, not lactate; humans make lactate, not ethanol or carbon dioxide.
Watch out Lactate is not a poison that must be got rid of straight away. It is carried in the blood and later used as a fuel, or made back into glucose in the liver.
glucoseglycolysisnet 2 ATP2 pyruvateNAD in the cellNADNADNADNADNADNADNADHNADH piles upNADHNADHNADHNADHNADHNADHNothing gives NAD back, so the store runs dryand glycolysis stops: no more ATP.NADNAD back to the store: glycolysis goes on2 lactatebloodMuscle: pyruvate becomes lactate, in the cytosol.Lactate is carried away in the blood; no extra ATP.2 CO22 ethanolbaking: carbon dioxidemakes the dough risebrewing: the ethanolis keptYeast: pyruvate loses a carbon as carbon dioxide, and ethanol is left. glucoseglycolysisnet 2 ATP2 pyruvateNAD in the cellNADNADNADNADNADNADNADHNADH piles upNADHNADHNADHNADHNADHNADHNothing gives NAD back, so the storeruns dry and glycolysis stops:no more ATP.NADNAD back to the store:glycolysis goes on2 lactatebloodMuscle: pyruvate becomes lactate,in the cytosol. Lactate is carriedaway in the blood; no extra ATP.2 ethanol2 CO2baking: carbon dioxidemakes the dough risebrewing: the ethanolis keptYeast: pyruvate loses a carbon ascarbon dioxide, and ethanol is left.

Glycolysis alone: each glucose turns two NAD+ from the store into NADH. Nothing gives NAD+ back, so the store runs dry and glycolysis stops.Muscle: pyruvate takes the hydrogen from NADH and becomes lactate. NAD+ goes back to the store, so glycolysis goes on.Yeast: pyruvate loses a carbon as carbon dioxide, and what is left takes the hydrogen from NADH, becoming ethanol. NAD+ goes back to the store.

Glycolysis with no oxygen. Watch the NAD+ in the store, then see what each cell does about it.

3.5.A The link to the Krebs cycle, and the cycle

The carbon leaves as carbon dioxide

With oxygen, pyruvate is carried from the cytosol into the matrix, the fluid inside the inner membrane of the mitochondrion. Two sets of reactions follow, one after the other.

First, each pyruvate (3C) is decarboxylated, which takes a carbon off as carbon dioxide, and oxidized: hydrogen is removed and accepted by NAD+, making NADH. The 2-carbon acetyl group that is left goes on into the Krebs cycle. Fatty acids and other macromolecules are broken down to acetyl groups too, and join the same route (chapter 10). All of this happens twice for each glucose, once for each pyruvate.

In the Krebs cycle, the acetyl group is added to a 4-carbon compound. The 6-carbon compound that results is taken round a cycle of reactions that ends by remaking the 4-carbon compound. Round the cycle, two carbons leave as carbon dioxide, and the compound is oxidized four times: three times the hydrogen goes to NAD+, and once to FAD, another hydrogen carrier, making FADH2. One ATP is made. The cycle turns twice for each glucose: 4 carbon dioxide, 6 NADH, 2 FADH2 and 2 ATP in all. You do not need the names of the compounds or the enzymes.

Each turn of the Krebs cycle takes in a 2-carbon acetyl group. Why does the cycle not fill up with carbon?

Watch out The Krebs cycle makes little ATP directly, only one a turn. Its main product is NADH, which the chain turns into most of the ATP.
Watch out The carbon dioxide you breathe out comes from the carbons of the fuel, not from the oxygen you breathe in. The oxygen goes to water, at the end of the chain.
cytosolmatrixintermembrane spaceinside a mitochondrionpyruvate3CCO2NADNADHacetyl2C acetyl groupLink reaction4C4C6C6C5C4C4C4CNADNADHNADNADHADPATPFADFADH2NADNADHCO2CO2Krebs cycletwo turns a glucosefor one pyruvatefor one glucoselinkKrebscarbon dioxide1224NADH1236FADH20012ATP0012 inside a mitochondrioncytosolintermembrane spacematrixpyruvate (3C)Link reactionCO2NADNADHacetyl2C4C4C6C6CreducedNADreducedNADATPreducedFADreducedNADCO2CO25C4C4C4CKrebscyclefor one pyruvatefor one glucoselinkKrebscarbon dioxide1224NADH1236FADH20012ATP0012
  1. Pyruvate goes into the matrix.
  2. Before the cycle: a carbon off as CO2 (decarboxylation), hydrogen to NAD+ (oxidation). The acetyl group goes on.
  3. Acetyl (2C) + a 4-carbon compound → a 6-carbon compound.
  4. Round the cycle: two decarboxylations, four oxidations (three to NAD+, one to FAD), one ATP.
  5. The 4-carbon compound is remade: a cycle.
  6. Two turns per glucose: all six carbons gone as CO2.
Eight steps to sortWhich kind of reaction is each step? What is lost or gained?Hydrogen removed from a3-carbon sugar in glycolysisNAD accepts hydrogen andbecomes NADHA carbon leaves pyruvate ascarbon dioxideGlucose gains a phosphate fromATPHydrogen removed from acompound in the Krebs cycleOxygen accepts electrons at theend of the chainA 6C compound loses a carbon ascarbon dioxide, on the wayround the cycleADP gains a phosphate, makingATPOxidationReductionDecarboxylationPhosphorylation

Pick a step (tap it, or Tab to it and press Enter), then pick the bin it belongs in.

0 of 8 sorted

Name the reaction: eight steps of respiration and four kinds of reaction. Which bin does each belong in, and what tells you?

3.5.B The electron transport chain

Down the chain

cytoplasmmitochondrionNADH and FADH2carried to the chainglycolysislactateno oxygenpyruvate oxidationKrebs cycleelectrontransportchainyou are herecytoplasmmitochondrionglycolysislactateno oxygencarbon dioxidewaterabout 28 to 30 more ATPwith oxygenyou are here

NADH is made in three places: glycolysis, the link reaction and the Krebs cycle. The electrons they carry all end up at the inner membrane of the mitochondrion, which is folded into cristae to give room for many copies of what happens there.

In that membrane sits the electron transport chain: a series of electron carriers, some of them proton pumps. A pair of electrons is passed from NADH to the first carrier, and the NADH is turned back into NAD+, which goes back for more hydrogen. That is the transfer of energy. FADH2 from the Krebs cycle hands its electrons on too, one carrier further along, so it pumps fewer protons.

The electrons are passed from carrier to carrier, each holding them a little more tightly than the one before, so they always move on and never go back. A little energy is released at each step, like a ball dropping down a staircase. Oxygen, at the very end, holds them most tightly of all (chapter 9). You do not need to name the carriers: “the chain” and “proton pumps” are enough.

The pumps use that energy to move protons (hydrogen ions) from the matrix, across the inner membrane, into the narrow intermembrane space. They pile up there, so there are far more protons in the space than in the matrix: a proton gradient. The membrane does not let them leak back, so the gradient is a store of energy, like water held behind a dam. Chapter 9 lets it out.

Watch out The chain does not make ATP; it makes a proton gradient. And “proton” here means a hydrogen ion, not a hydrogen atom.

Electrons pass along the chain of carriers, releasing a little energy at each step. What is that energy used for?

proton (a hydrogen ion)electrona small electron carrierouter membraneintermembrane spaceinner membranematrixNADHfrom glycolysisNADunloaded: NAD againNADHfrom the link reactionNADHfrom the Krebs cycleto oxygen(chapter 9)FADH2from the Krebs cycleproton pumpsmany protons: a proton gradientfew protons in the matrix proton (a hydrogen ion)electrona small electron carrierouter membraneintermembrane spaceinnermembranematrixreducedNADfromglycolysisNADunloaded:NAD againreducedNADfrom thelink reactionreducedNADfrom theKrebs cycletooxygenreducedFADfrom the Krebs cycleproton pumpsmany protons:a proton gradientfew protonsin the matrix
  1. NADH from all three stages arrives at the inner membrane.
  2. It hands a pair of electrons to the first carrier, and becomes NAD+ again.
  3. Each step down releases a little energy, used to pump protons. FADH2 hands its electrons on one carrier later.
  4. Protons pile up in the intermembrane space: a proton gradient.

3.5.B Chemiosmosis and ATP synthase

Protons through the turbine

Protons can return to the matrix through one protein only: ATP synthase. It spans the inner membrane, with a channel through it, a part that turns, and a head in the matrix where ATP is made.

As protons flow down their gradient through the channel, they turn the rotor. The turning changes the shape of the head, which joins ADP and phosphate into ATP. ATP synthase couples the energy released by the flow of protons to the phosphorylation of ADP. Making ATP with the energy of a proton gradient across a membrane is called chemiosmosis. In cellular respiration the whole process, the chain with the synthase, is called oxidative phosphorylation.

The chain needs one more thing at its end. Oxygen is the terminal electron acceptor: it takes the electrons from the last carrier, and protons from the matrix, and becomes water, called metabolic water. That empties the chain so that electrons keep flowing, and taking protons from the matrix helps to keep the gradient steep. About 26 to 28 of the 30 to 32 ATP from one glucose are made here, by ATP synthase.

So ATP is made in two ways. A little is made directly, in glycolysis and the Krebs cycle, when a phosphate is passed from a substrate to ADP: substrate-level phosphorylation. The rest is made by ATP synthase, with energy brought to the chain by the carriers: about 2.5 ATP for each NADH and 1.5 for each FADH2. The ledger after the film adds it up, stage by stage.

Not every cell wants all of its energy as ATP. If protons leak back across the inner membrane by another route, without passing through ATP synthase, the electron transport chain still runs but the energy of the gradient is released as heat. This is uncoupling. Mammals that need to stay warm, such as newborns and hibernating animals, use it in their brown fat: their cells respire a lot of fuel and make a lot of heat.

This is not only a mitochondrion. The same kind of chain, with a proton gradient and ATP synthase, works in the thylakoid membranes of chloroplasts and in the plasma membrane of prokaryotes. Aerobic prokaryotes end their chain on oxygen, as we do. Anaerobic prokaryotes end it on other molecules instead, so the chain still runs and the gradient still makes ATP without oxygen.

How we know (extra) In 1961 Peter Mitchell suggested that the chain and the making of ATP were linked by a gradient of protons, not by some unknown high-energy chemical. Most biochemists doubted it for years. Experiments in the 1960s and 1970s, in which membranes given an artificial proton gradient made ATP, won them over, and he received the Nobel Prize in Chemistry in 1978.
Watch out ATP synthase is not a pump: protons flow through it, down their gradient, and that flow drives it. The pumps are in the chain.
Watch out Oxygen does not become carbon dioxide. It ends up in water. The carbon dioxide came from the fuel’s carbon, in the link reaction and the Krebs cycle.

A cell’s mitochondria run out of oxygen. What happens to the electron transport chain?

proton (a hydrogen ion)electronintermembrane spaceouter membranematrixATP synthaseADPphosphateATPoxygenwaterNADHchemiosmosisthe gradient drives ATP synthase:about 26 to 28 ATP a glucose proton (a hydrogen ion)electronouter membraneintermembrane spaceATP synthasematrixADPphosphateATPoxygenwaterreducedNADchemiosmosisthe gradient drivesATP synthaseabout 26 to 28ATP a glucose
  1. One way back for the protons: ATP synthase.
  2. Protons flowing down the gradient turn it, and their energy joins ADP and phosphate: ATP.
  3. Oxygen takes the electrons and protons at the end, making water.
  4. Chemiosmosis: the gradient makes ATP, and oxygen keeps the chain flowing.
Watch the protons pile up, then turn the oxygen off and see what stops first. Then open the leak.
substrate-levelphosphorylationATP made directlycarried tothe chainoxidativephosphorylationthrough ATP synthasein all4 ATP10 NADH2 FADH2about 26 to 28 ATPabout 30 to 32 ATPfrom one glucose, with oxygencytoplasm2 ATPnet: 4 made, 2 used2 NADHabout 3 to 5 ATP1.5 to 2.5 eachglycolysismatrixnone2 NADHabout 5 ATPabout 2.5 eachpyruvateoxidationmatrix2 ATP6 NADHabout 15 ATPabout 2.5 each2 FADH2about 3 ATPabout 1.5 eachKrebs cycleGlycolysis makes its NADH in the cytoplasm, and passing its electrons into themitochondrion can cost some of their worth. Older books say 36 to 38 ATP: they count 3a NADH and 2 a FADH2.with oxygenabout 30 to 32 ATPglycolysis: 2in the mitochondrion: about 28 to 30without oxygen2 ATPpyruvate becomes lactate: no more ATPglycolysis: 2So without oxygen a cell uses about 15 times the glucose for the same ATP.
Every stage against the ATP it makes, directly or through the chain. Tap a stage to see it on the map.

3.5.A Lipids and carbohydrates as fuels

Fat or sugar?

Glucose and fatty acids are the two main fuels, and they are not equal. Lipids release about twice as much energy per gram as carbohydrates when they are respired: about 37 kJ in each gram against about 17.

They also take different routes. Carbohydrate goes through glycolysis, so it can be respired with oxygen or without. A fatty acid is cut into 2-carbon acetyl groups, which enter the Krebs cycle at once. Glycolysis is the one stage that works without oxygen, and a fatty acid never goes through it: no oxygen, no use for fat. Proteins can be used too: amino acids lose their nitrogen and the carbon skeleton joins the pathway as pyruvate or as an acetyl group.

In practice, muscles use fat for long, steady work, such as walking, a gentle jog or a migrating bird in flight, and carbohydrate for hard, fast work, when oxygen is short. That is why fat is the long-term store and glycogen the quick one (unit 1).

Gram for gram, fat releases about twice the energy of carbohydrate when respired. Why?

Watch out Fat cannot be used anaerobically: fatty acids enter at the Krebs cycle, which needs the chain, which needs oxygen. A sprint runs on carbohydrate.
OHOHHOHOHHHHOHCH2OHOHOglucosepalmitic acid, a fatty acidBy massC 40%H 7%O 53%C 75%H 13%O 12%carbonhydrogenoxygenEnergy released when a gram is respiredcarbohydrateabout 17 kJ g−1lipidabout 37 kJ g−1mitochondrioncytosolglucoseglycolysispyruvateno oxygen: lactate2 ATPlinkacetylfatty acidno oxygen: no routefat never enters glycolysiscut into 2Cacetyl groups8 from palmitic acidKrebs cycleand the chainoxygenneededOxygen used to respire a gramcarbohydratelipidabout 2.7 times as much OHOHHOHOHHHHOHCH2OHOHOglucoseBy massC 40%H 7%O 53%palmitic acid, a fatty acidC 75%H 13%O 12%carbonhydrogenoxygenEnergy released when a gram is respiredcarbohydrate: about 17 kJ g−1lipid: about 37 kJ g−1cytosolglucoseglycolysispyruvateno oxygen:lactate, 2 ATPfatty acidno oxygen:no route: itnever entersglycolysismitochondrionlinkacetylcut into 2Cacetyl groups8 from palmitic acidKrebs cycleand the chainoxygen neededOxygen used to respire a gramcarbohydratelipidabout 2.7 times as much

What is in them: glucose is about half oxygen by mass, palmitic acid about one eighth. A gram of lipid releases about 37 kJ, a gram of carbohydrate about 17.How they are respired: glucose goes through glycolysis in the cytosol, so it has a route without oxygen. A fatty acid is cut into acetyl groups in the mitochondrion, which needs oxygen. A gram of lipid uses about 2.7 times as much oxygen.

Glucose and a fatty acid, side by side: what they are made of, and how each is respired.
Every stage in one film. Switch the fuel to a fatty acid, or the cell to yeast, then cut the oxygen.
ribosomesDNAcytoplasmmitochondrionits electronsare passed in2 pyruvatematrixinner membranefolded into cristae2 acetyl CoAglucose2 ATPnet: 2 used, 4 made2 NADHglycolysisno more ATPlactatein yeast: ethanoland carbon dioxideno oxygen2 carbon dioxide2 NADHpyruvate oxidation4 carbon dioxide2 ATP6 NADH2 FADH2Krebscyclepumps protons into theintermembrane spaceoxygenwaterelectron transportchainprotons flow backabout 26 to 28 ATPATP synthaseintermembrane spaceIn all, from one glucoseglucose + oxygencarbon dioxide + wateraerobic respirationabout 30 to 32 ATPglucoselactateanaerobic, in humans2 ATPglucoseethanol + carbon dioxideanaerobic, in yeast2 ATPribosomesDNAcytoplasmmitochondrion2 pyruvatefolded innermembraneglucose2 ATPglycolysisno more ATPlactatein yeast: ethanoland carbon dioxideno oxygen6 oxygen6 waterabout 28 to 30 more ATP6 carbon dioxidewith oxygenIn all, from one glucoseglucose + oxygencarbon dioxide + wateraerobic respirationabout 30 to 32 ATPglucoselactateanaerobic, in humans2 ATP
The whole map again, with the word equations and the totals. Tap a stage to go over it.

02Dry lab: Respirometer: peas and dough

Germinating peas in a sealed tube use oxygen, and a colored drop creeps along a scale as they do. Change one thing, read the drop each minute, and work out how fast the peas respire. Or switch to dough and race five sugars.

Your questionHow does one factor change the rate at which germinating peas take up oxygen? Start with temperature. Then try the dough: which sugar makes yeast respire fastest?
What you can changeOne independent variable at a time: the water bath's temperature, how long ago the seeds were soaked, the mass of seeds, or whether the potassium hydroxide is in. In dough mode, the sugar. Everything else stays fixed unless you change it on purpose.
What to recordThe drop's position on the scale every minute, in mm, for the peas and for the glass-bead tube beside them. Then the rate in mm³/g/min, three or more trials at each value, a mean and a standard deviation, and a graph.
glucosefructosesucrosestarch050100150none30 °C0:00min : s050100150200050100150200colored dropstart25 °C2.03 g0:00min : sbalanceclockseedsglass beadsseedsglass beads12341capillary2screw clip3potassium hydroxide4wire gauzepotassium hydroxide is corrosivethe bath is under a dark cover
  1. Yeast dough: five tubes, one sugar in each, read against a ruler every 5 minutes.
  2. Germinating seeds over potassium hydroxide, with a glass-bead tube beside them as the control. Both drops sit at the start mark: read each one every minute.
Run it on the page: set it up, press Start, read the drops each minute, and the table and graph fill in.

Open the bench full screen: the apparatus, its controls, your readings and the graph then fit on one screen.

Find out

  • Temperature. Run 5 to 40 °C in 5 °C steps, with 3-day seeds and 2.0 g, three trials at each. Then try 45 °C: what happens to the drop as the run goes on?
  • Waking up. At 25 °C, run dry seeds, then 1 to 5 days since soaking, then boiled seeds.
  • Per gram. At 25 °C, run 1 to 6 g of seeds. The table gives the rate for the whole tube: work out the rate per gram for each mass too, and plot both.
  • No potassium hydroxide. Run 3-day mung beans with it in, then with water in its place.
  • Which apparatus? At 25 °C, run 3-day seeds with the capillary and its drop, then with the U-tube manometer and its control tube. Which reads more precisely, and why? (A third, an insect over soda lime with a bubble in a pipette, is in the check questions.)
  • The clip. At 25 °C, leave the clip open for 0, 5 and 10 minutes before starting. Watch the bead tube's drop.
  • Five sugars. Switch to yeast dough at 30 °C and run the rack three times, so you have three tubes of each sugar, over 30 minutes.

Record

  • Every run's settings: what you changed, and everything you kept the same.
  • Both tubes' readings every minute, the seeds' and the glass-bead tube's, in the units your apparatus reads. For the dough, each top every 5 minutes.
  • Change one thing at a time.
  • Three or more trials at each value, then a mean and its standard deviation.
  • The bench’s graph plots each reading as it is taken. Work out each rate yourself, then press Calculate to check it.
  • For the means and their spread, use Graph your results under the lab, or download the CSV and plot it yourself.
  • The bench keeps each variable's results: choose it again to see them.
  • When you are done, choose one investigation and write it up: the question, the method, a table, a graph with error bars, and a conclusion that cites your data.

With the potassium hydroxide taken out, the drop beside the mung beans hardly moves. Why?

In the first two minutes the drop beside the glass beads moved 10 mm away from its tube, then stopped. What does that tell you?

Sucrose dough rose almost as fast as glucose dough, but starch dough barely beat dough with no sugar. Why?

03Check

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05Teacher resources

The guide as three one-hour lessons. Each has a plan to print (the timings, what to project and where to stop, the activities with what they need, what students get wrong) and slides to teach it from, in the same order, with the teacher's moves in the notes.

LessonCoversFocusChaptersLesson planSlides
1From glucose to pyruvate 3.5.A60 min Respiration as a controlled release of energy, glycolysis and the carriers, and the respirometer 1, 2, 3, 4, 5, dry lab
2The mitochondrion 3.5.A, 3.5.B60 min The Krebs cycle, the electron transport chain and chemiosmosis, built with a class model and finished on the ATP ledger 7, 8, 9
3Without oxygen, other fuels, free response 3.5.A, 3.5.B60 min Fermentation, fuels and a timed free response on respiration data 6, 8, 10, dry lab

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How this was made. The course structure, the activities and the teaching choices come from my own classroom, my presentations and my years of teaching IB and AP Biology. I use Claude, an AI, to help write and build the interactive guides, lesson plans, slides and practice questions, and to draw the diagrams and code the interactives. I direct the work and review it against the IB and AP specifications before it goes live.