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Introduction
Photosynthesis and respiration are fundamental biological processes photosynthesis produces the carbohydrate and oxygen that plants animals and many microbes rely upon respiration releases the energy stored in organic molecules in a usable form Photosynthesis Respiration Click on the button to select the topic
Photosynthesis
Select topic by clicking on the button General introduction Photosynthesis and chloroplast structure LightLight-dependent and independent stages Factors affecting photosynthesis Measurement of photosynthesis Extension material Summary
Introduction
Photosynthesis is probably the most important biological process process. Without it, where would organisms get the carbohydrate and oxygen they need for respiration? With the exception of chemosynthetic bacteria, all other organisms ultimately depend upon plants and their photosynthetic activity. So, the ultimate source of the energy used by most , gy y organisms is the sun, and photosynthesis forms the basis of most food chains.
You may be familiar with the terminology already, but: Autotroph They are self-feeders, using simple selfinorganic molecules to build complex organic molecules. Uses light energy to fix CO2 into sugar sugar. Uses chemical energy to fix CO2 into gy sugar, e.g. nitrifying bacteria. nitrifying Other feeders that must have complex organic molecules like carbohydrates, lipid and p p protein in their food.
The ultimate source of energy is the sun! Plants harvest solar energy and convert it i t Pl t h t l d t into chemical potential energy of photosynthetic products. These are a source of energy for the plants themselves and for the heterotrophs that feed off the plants. Respiration converts the chemical potential energy of organic matter into chemical p g potential energy of ATP. gy
Solar energy
Autotrophs trap light energy, p g gy converting it into chemical energy of organic molecules
Heterotrophs animals, fungi and bacteria g consume organic nutrients carbohydrate, lipid and protein
Respiration provides usable energy for cells ATP produced and stores energy in its bonds Heat energy lost/released because processes are inefficient Work uses the energy contained in the bonds of ATP
There are two sets of reactions in photosynthesis p y LightLight-dependent stage light energy is trapped by the photosynthetic pg pigments used to produce ATP and to split water into hydrogen and oxygen takes place in the membranes of th b f the chloroplasts LightLight-independent stage light is not necessary (hence used to be called the dark stages/reactions) energy trapped in the lightlight-dependent stages is used to reduce CO2 into sugar takes l t k place i th li id in the liquid component (stroma) of the chloroplast
Chloroplast structure
the chloroplast is an organelle found in photosynthetic eukaryotic cells it is where photosynthesis takes place light energy used to fix CO2 into sugar basic chloroplast structure was covered in Unit 1, so you should already be familiar with it
under the light microscope chloroplasts i hl l appear as biconvex discs (3(3-10 m diameter) little detail is evident
electron microscope has better resolution and magnification therefore more detail th f d t il can be seen
TEMofColeusblumei chloroplast.Starchgranulesand thylakoidsarrangedintogranaareclearlyvisible.Public domainimage:LouisaHowardandCharlesDaghlian
chloroplast are surrounded by a double membrane/envelope flattened sacs or thylakoids run through the internal solution or stroma the th sacs are stacked in t k di places. Here they are called grana
they are the site of photosynthesis. Grana trap light energy which is i used t fi CO2 i th d to fix in the stroma
Starch grain. Starch is insoluble and so it does not affect the water potential. Thylakoid. Flattened fluid-filled sac. The membranes hold pigments and carriers. Granum. A stack of thylakoids. Light-dependent stage takes place here. Stroma. Contains the enzymes that fix CO2. lightindependent stage.
Additional points
In the stroma are starch grains and lipids It also has small 18nm/70S ribosomes a loop of DNA What is the significance of each of these?
Starch grains and lipid droplets are for storage of photosynthetic product
they are osmotically inactive and so can be stored without affecting the water potential
LightLight-independent stage
Light is t Li ht i not needed ( d d (used d to be called the dark stages/reactions). g ) Energy trapped in the lightlight-dependent stages is used to reduce carbon dioxide to sugar. Takes place in the stroma stroma.
Click to select topic on aspects of the light-dependent lightand independent stages of photosynthesis LightLight-dependent stage Absorption and action spectra Cyclic photophosphorylation Non- li h t h Non N -cyclic photophosphorylation h l ti Photolysis of water LightLight-independent stage Effect of photosynthesis on Calvin cycle intermediates Mechanism of ATP production Sequencing the Calvin cycle
LightLight-dependent stage
Light energy is trapped by photosynthetic pigments
organic molecules which absorb certain wavelengths of light, but t th li ht b t not others absorbed light is used to produce usable energy light that is not absorbed is transmitted or reflected
Most of the pigment in the chloroplast is chlorophyll, of which there are two basic types
chlorophyll a and b
Chlorophyll
absorbs blue and red light strongly transmits or reflects green light
hence the green colour of most leaves
Accessory pigment includes various forms of chlorophyll a, chlorophyll b and others (carotene, xanthophyll, etc.). p y ) Pigments are arranged into light-harvesting clusters lightknown as photosystems. In photosystems, several hundred pigment molecules absorb and then funnel or feed the light they absorb into i t a reaction centre which contains primary ti t hi h t i i pigment.
sometimes the main light harvesting pigment is referred to as antenna pigment
P700 is at the reaction centre of PS I, whilst P680 is at the reaction centre of PS II.
The light-dependent stages include the production of lightATP and splitting of water (photolysis) to produce H+ ions ions The H+ ions combine with NADP, reducing it. The Th production of ATP can be either cyclic or nond ti f b ith li noncyclic, depending upon the pattern of electron flow in the photosystems in cyclic, they end up where they started from. in non-cyclic they dont! non-cyclic don t!
Chlorophyll absorbs light with wavelengths 400-700nm p y g g 400photosynthetically active radiation or PAR for short
Relative absorbance a e
400
500
700
How are different wavelengths absorbed by the different photosynthetic pigments? Chlorophyll a and b have peaks of absorption in the blue and red regions of the spectrum with very little absorption in the green region. In contrast, carotene has a broad band of absorption from , p blue to green and extends the wavelengths that are available to the plant. It is likely that accessory pigments also protect the chlorophyll when light is very strong. What effect does wavelength (light quality) have on the rate ff ( ) of photosynthesis? Absorption spectra should not b confused with th action Ab ti t h ld t be f d ith the ti spectrum, which is a plot of photosynthesis against wavelength. wavelength Not surprisingly there are peaks of action surprisingly, (photosynthesis) where there are peaks of absorption!
Why do the leaves of trees turn yellow-orange in autumn? yellowThe Th magnesium i th chlorophyll i withdrawn so th i in the hl h ll is ithd the chlorophyll loses its green colour and the colour of the accessory pigment can be seen seen.
Cyclic photophosphorylation
Cyclic photophosphorylation only involves PSI Light is absorbed by pigments and passed to P700
This Thi excites an electron which moves t a hi h it l t hi h to higher energy state and are emitted from the chlorophyll a The electrons pass to an electron acceptor (FD) and then back to P700 via another electron acceptor (PQ) and a series of electron carriers which make up the electron transport chain or ETC As electrons travel along the ETC energy is lost, enough energy to generate ATP from ADP and inorganic P (Pi) i i
FD
PQ E
e-
C P700 PSI
ADP+ Pi ATP
NonNon-cyclic photophosphorylation
This involves both PSI and PSII. The process has become known as the Z-scheme. ZLight is absorbed by both photosystems, electrons excited and emitted from the primary pigments and transmitted to the electron acceptors and transferred t itt d t th l t t dt f d along the ETC. P700 gets its electrons from PS II whilst P680 gets its electrons from the splitting of water.
PS II has a water-splitting enzyme water-
As in cyclic photophosphorylation
energy is released used to generate ATP from ADP and Pi
FD NADP + 2H+ PQ H2O 2eADP ADP+ Pi 2H+ O2 P680 PSII ATP E 2eT C P700 PSI = reduced NADP
Cyclic Non-cyclic
Non-cyclic PSI & PSII Light, ADP, Pi, water, NADP ATP, NADPH2, O2 Water splitting enzyme present associated with PSII
Light, ADP, Pi
Products
ATP
Other points
Photolysis of water
Photolysis of water is vital to photosynthesis. PSII includes a water-splitting enzyme: waterH2O ------> 2H+ + 2e- + O2 ------> The H+ ions combine with electrons from PSI and NADP to produce reduced NADP this is then used together with ATP in the light-independent stages to lightreduce CO2 into carbohydrate. carbohydrate
Photolysis of water and production of reducing power can be demonstrated by the Hill reaction DCPIP can reaction. substitute for NADP, receiving electrons and becoming reduced: g Oxidised O idi d DCPIP blue H2O Reduced DCPIP R d d colourless O2
LightLight-independent stage
The light-independent stage takes place in the lightstroma of the chloroplast
contains the necessary enzymes key enzyme is ribulose bisphosphate carboxylase (RUBISCO)
CO2 combines with ribulose bisphosphate to produce 2 molecules of glycerate phosphate (GP)
also known as phosphoglyceric acid/phosphoglycerate and PGA
GP is not a sugar, but is reduced into one using the energy and reduced NADP produced during the lightlightdependent stages
Calvin cycle:
CO2
RuBP
Unstable intermediate
GP
GP
redNADP redNADP ADP ATP Triose P NADP NADP ATP ADP+Pi Triose P
Most of the triose produced is used to regenerate the CO2-acceptor ribulose bisphosphate. However
one turn of the Calvin cycle generates 1 spare fixed carbon two turns generate 2 spare fixed carbons (could be used to make acetyl CoA and then lipid or amino acids) three turns generate 3 spare fixed carbons or one spare triose six turns generate 6 spare fixed carbons or two spare triose enough for one hexose
Clearly levels of RuBP, GP and TP can fluctuate depending upon the prevailing conditions and the amount of photosynthesis occurring. occurring
Ribulose bisphosphate
The levels of RuBP are affected by a number of conditions
decrease when photosynthesis is rapid consumed as CO2 is added to it regenerated by th C l i cycle so l t d b the Calvin l levels reach a l h steady state
Paradoxically, the high levels of photosynthesis under these conditions reduce the amount CO2 available and increase the amount of O2
makes it more likely that O2 occupies the active site of the enzyme
inhibits photosynthesis increases photorespiration
What type of enzyme inhibition is responsible for photorespiration? Explain. Competitive inhibition Oxygen and carbon dioxide compete for the active site What effect might photorespiration have upon crop p productivity? y Reduce it Photorespiration consumes RuBP, wasting the p , g energy that was used to make it. It also needs regenerating, this uses more energy Note: despite being called photorespiration, it is not respiration as it produces no ATP whatsoever.
Consider the example of a plant that has been in wellilluminated conditions and then the light is switched off.
GP
TP
Inthelight
Inthedark
Inthelight
In the light:
concentrations of GP and TP reach a steady state.
Concentra ationofGP PandTP
GP TP
GP is made when CO2 is added to RuBP by the enzyme RUBISCO. TP is made from GP using ATP and reduced NADP from the light-dependent stage.
In the dark:
GP is still made (RUBISCO is still active) but it is not used to active), make TP
ATP and reduced NADP are not available. levels of GP increase and TP decrease. Note GP plateaus when all of the RuBP is consumed.
Inthelight
Inthedark
Inthelight
Key theory/concept
the basis of the mechanism for ATP production in chloroplasts also the mechanism for ATP production in respiration
NADP
NADPH + H+
ADP + Pi
H+ ATP
H+ O2 H+
H+ H+ H+ H+
In a nutshell:
transfer of electrons through the electron transport chain releases energy h i l
this energy is used to pump protons (H+ ions) out of the stroma and into the thylakoid space generates a region with a high concentration of H+ ions
photolysis of water also generates H+ ions H+ ions then flow back into the stroma
through a membrane protein
contains an ATP synthase so generates ATP
Side view of the special lollipop S f vessel Port f P t for injecting the 14CO (as 2 14CO ) NaH 3
Front view of the special lollipop f vessel Port for injecting the 14CO (as 2 NaH14CO3)
Li ight source
Li ight source
Calvin reasoned that 14C label would appear first, and in the largest amount, in the first products of photosynthesis. h t th i Label would then be passed on to the subsequent products of photosynthesis in the order in which they were produced. The biochemical sequence he identified is now commonly referred to as the Calvin or C3 cycle.
He separated and identified products of photosynthesis paper chromatography to separate autoradiography to locate and identify (chromatograms placed on photographic ( h t l d h t hi film/paper. Radioactivity caused fogging when it was developed) He produced graphs that clearly show the sequence of 14C fixation during photosynthesis
Time
Note: N t sometimes th C l i cycle i called th C3 cycle b ti the Calvin l is ll d the l because the first stable product is glycerate phosphate
Click on the button to select the limiting factor of photosynthesis: Light intensity and wavelength Carbon dioxide C b di id Temperature Water Nutrients Interaction of factors Controlling p g productivity y
At LSP, some factor other than light will be limiting, e.g. e g temperature or CO2?
Light intensity
Rate +
_ Light intensity
Wavelength
Light is absorbed by the plant's photosynthetic pigments Chlorophyll absorbs light with wavelengths 400400700nm (so-called Photosynthetically Active Radiation (soor PAR for short) Not all wavelengths are absorbed equally red and blue are absorbed strongly green is absorbed weakly most is reflected or transmitted Why do plants appear green?
Relative absorbance a e
400
500
700
photosynthesis
Photosynthesis is proceeding at maximum rate. Some factor other than CO2 is limiting g photosynthesis.
CO2 concentration
As much CO2 is being produced by respiration as is being consumed by photosynthesis. Net photosynthesis = zero.
Under most circumstances CO2 limits photosynthesis Note: when plants are water stressed they close stomata
to li it t limit water loss t l but it also limits CO2 uptake therefore affects photosynthesis
Temperature
Photosynthesis is enzyme controlled. There are two important enzymes involved in photosynthesis:
ribulose bisphosphate carboxylase (RUBISCO) waterwater-splitting enzyme of PSII
Photosynthesis Doubling of rate per 10oC rise in temperature up to the optimum. Q10 of 2.
Optimum temperature
Photosynthesis proceeds at fastest rate.
Denaturation
Irreversible damage to enzymes means rate falls rapidly to zero.
Temperature / oC
Water
An adequate supply of water is needed:
water is a substrate in photosynthesis
however, most of the plants water requirements are to replace water lost by transpiration.
In salt marshes there is plenty of water but plants suffer physiological drought:
plants cannot generate a sufficiently low water potential to absorb all of the water needed. they have xerophytic adaptations.
Nutrients
Plants need mineral nutrients for photosynthesis. May be needed:
to make chlorophyll for photosynthetic enzymes
Key minerals are: y Magnesium in chlorophyll i hl h ll Iron for chlorophyll ETC components
Interaction of factors
Photosynthesis is affected by a number of factors, the one in least supply limits the overall process Consider interaction of light intensity temperature CO2 concentration
photosynthesis
Increasing light intensity increases photosynthesis until saturation is reached and some other factor becomes limiting Raising the temperature raises the rate of photosynthesis h t th i doubling it? Q10 = 2 Raising CO2 raises the rate of photosynthesis f it is the substrate
At low light intensities, increasing the temperature has no effect on photosynthesis gradient is the same and so the Q10 must be 1 Photochemical reactions are not affected by temperature (Q10 = 1) t t EnzymeEnzyme-controlled reactions are affected by temperature (Q10 = 2) this observation led early plant physiologists to suggest that photosynthesis consisted of lightlightdependent and light-independent stages/processes light-
Carbon dioxide
Providing extra CO2 should boost photosynthesis If CO2 concentration > 0.5%, ,
can cause stomata to close have the opposite of the desired ff t! d i d effect!
Shade on bright days Provide the optimum intensity and duration for the crop
Temperature
Raise temperature by p y
limiting air movement transparent
lets light in, stops heat loss
Water/humidity
Automated watering: g
hydroponics
supply water and nutrients
sprinklers
to water
humidifiers
maintain humid atmosphere keep stomata open
The control of abiotic factors is not cheap. Farmers must balance the extra costs of glasshouse cultivation with the extra revenue generated. If it costs more to increase yield than the extra y y yield g generates in income for the farmer, he will soon go out of business. Major costs j The structure
glass/poly tunnel
Running costs g
heat, light, etc.
Pest control
conditions in glasshouses favour disease
So, from the following equation th t summarises S f th f ll i ti that i photosynthesis 6CO2 + 6H2O + light energy C6H12O6 + 6O2 Which are the substrates? CO2 and water d t Which are the products? glucose/carbohydrate and O2 l / b h d t d
Traditionally, O2 evolution or CO2 consumption would be used to measure photosynthesis Suggest how each could be measured CO2 use pH change
CO2 is an acid gas remove it and pH i increases
O2 evolution bubble count collect gas evolved and measure volume oxygen electrode
Oxygen production can be measured using a photosynthometer O2 collects at the bottom of the flared capillary tube bubble of oxygen is drawn into the capillary tube by the syringe length measured against the scale
flaredendcapillarytube thermometer O2 bubble
syringe scale Ligh htsource shootofpondweed,e.g.Elodea pondwaterordiluteNaHCO pond water or dilute NaHCO3 beakerofwater/waterbath /
The distance moved is proportional to the rate of p photosynthesis, more specifically to the net oxygen y , p y yg produced by photosynthesis! What other information is needed to calculate the volume of oxygen produced and the actual rate of oxygen production? internal diameter/radius of the capillary tube crosscross-sectional area of the capillary tube time What formula would you apply to measure the rate of oxygen production? r2 x distance moved / time Do any factors need controlling so that we are confident in our estimate of photosynthesis? Working in pairs/small groups, describe and explain the effect of each factor and its control.
Factor Temperature
Effect if uncontrolled
Control measure
Light
Carbon dioxide
Other
Factor Temperature
Effect if uncontrolled Temperature influences photosynthesis increases if temperature rises but does not exceed the optimum. p Thermal expansion of gas. Variation in light results in variation in photosynthesis photosynthesis. Note: if light close to photosynthometer, it is likely to heat it. it Variation in CO2 produces variation in photosynthesis. CO2 is usually limiting, but if too high can cause stomatal closure. Amount of plant material Age of material Physiological state well watered or wilted healthy or chlorotic
Control measure Water bath or room temperature, with frequent checks on temperature. Choose optimum.
Light
Exclude extraneous light blinds to windows, room lights windows lights. Standardise wattage and distance from light. Choose saturating light. Immerse in HCO3- solution or allow HCO3- solution to equilibrate with air. Choose saturating CO2.
Carbon dioxide
Other
Hopefully, in addition to control of temperature, light and CO2 availability, you have decided to control the amount of plant material and its age/condition. A t f l t t i l d it / diti duplicate apparatus, but without the pondweed, could also be included as a control control.
It is possible to investigate the effect of varying temperature or light or CO2 whilst keeping the other factors constant constant. Suggest how you could vary the factor: Temperature
Use a water bath
thermostat control thermometer to check
Light
Vary distance from light
inverse square law I = 1/d2
Room temperature?
Light quality () (
Prism to split white light Monochromatic filters
need to check intensity with light meter
CO2
Hydrogen carbonate solution
aquatic p q plant
immerse in solution
terrestrial plant
solution equilibrates with l ti ilib t ith atmosphere
Accessory pigments and algal zonation Leaf structure adaptations to photosynthesis and gas exchange Sun and shade adaptations of leaves
Accessory pigments
You already know that: Pigments are classified as either primary pigments (P680 and P700) or accessory pigments Accessory pigments are often present but their colour is i masked b th d k chlorophylls k d by the darker hl h ll
xanthophyll (pale yellow) and carotene ( t (orange) (orange)
Some other accessory pigments are seen more easily in the algae
seaweeds show a zonation of types with depth
Green algae
possess chlorophyll need large amounts of red and blue light green algae near the surface
Brown algae B l
accessory pigment is coloured brown (fucoxanthin) red and blue light absorbed more efficiently brown algae at depths 10-25m g p
Red algae
accessory pigment is coloured red (phycoerythrin) absorb and use the mainly blue light found at depth red algae at greater depths
Leaf structure
The leaf is the main photosynthetic organ Basic leaf structure was covered in Cells, Exchange and Transport. Briefly: It is broad and thin
large surface area short diffusion path
It has chlorenchyma thats cells with p , p y chloroplasts, i.e. photosynthetic tissue Stomata are present for gas exchange
it must absorb light it must absorb carbon dioxide and remove waste oxygen
contains pigments large surface area thin leaf minimises diffusion distances gas exchange pores or stomata are present vascular tissue present to transport water and organic nutrients
it must have a water supply and be able to export organic nutrients it has made
upper epidermis
parenchyma
phloem
palisade mesophyll
long cylindrical cells, fewer cell walls for light to arranged at right angles to pass through and be upper epidermis attenuated thin cell walls long narrow air spaces between the cells numerous chloroplasts restricted to a thin layer of cytoplasm cyclosis low diffusion resistance large surface area for gas exchange rapid photosynthesis large vacuole pushes chloroplasts to periphery prevents damage due to excessive light
spongy mesophyll
irregular shape and irregularly packed p present, but chloroplasts p fewer in number moist cell walls help with gas exchange
large air spaces and large surface area for gas g exchange photosynthesis but less p y than in palisade create humid internal atmosphere and so increases chance of water loss from the leaf
xylem
cells elongate, dead h ll hollow cylinders j i d li d joined end-toend-to-end walls strengthened with lignin pits present
fine tubes aid water flow rather like pipe sections th lik i ti prevents collapse during transpiration allows movement of water and dissolved minerals out of xylem i l t f l
phloem
translocates organic nutrients cells elongated thin cytoplasm sieve plates connect the cells
this is an active process, requiring energy fine tubes help flow low resistance to flow form long tubes for transport and allow free llow movement mo ement from one sieve tube to another
stomatal structure
guard cells surround th d ll d the stoma (singular) each guard cell is sausagesausageshaped and contains chloroplasts cell wall facing the stoma is thicker than the rest of the cell wall during opening, the guard cell absorbs water thick inner cell wall resists stretching so cell becomes more sausage-shaped sausagesausage-shaped guard cells contain chloroplasts
Sun and shade-adapted plants shadeSome plants are adapted to bright light whilst others are adapted to low light. Even on the same plant:
some leaves will be adapted to bright light
e.g. those i th crown and e.g. th in the d
some to shade
e.g e g. those at the base e.g.
Photosynthesis
Sun leaf note the relatively high compensation point and saturation point
Shade leaf note the relatively low compensation point and saturation point. Also the dip at high light intensities
SunSun-adapted Smaller surface area Leaf blade is thicker Palisade layer may b P li d l be 23 cells thick Fewer chloroplasts Less accessory pigment High LCP High LSP
Shade-adapted ShadeSurface area greater Thinner leaf blade Palisade layer only 1 P li d l l cell thick More chloroplasts More accessory pg pigment Low LCP Low LSP (may even suffer at the highest intensities)
Summary
Photosynthesis is an y important biological process
it produces the carbohydrate and oxygen relied upon by many organisms takes place in chloroplasts
return to P700 via electron transport chain lost l t energy li k d t ATP linked to production
nonnon-cyclic photophosphorylation
involves both PSI and PSII light absorbed by PS
highhigh-energy electrons emitted
LightLight-dependent stage
takes place in the thylakoids
light energy is harvested used to produce ATP and reducing power (reduced NADP)
cyclic photophosphorylation
involves light-harvesting lightclusters called photosystem I P700 as primary pigment light absorbed passed to P700 high energy electrons emitted
PSII has water-splitting waterenzyme: H2O 2H+ + 2e- + O2 electrons from water replace those lost by P680 H+ pass to NADP, reducing it
LightLight-independent stage
takes place in the stroma uses ATP and reducing power from the lightlightdependent stage In the Calvin cycle, CO2 is added to ribulose bisphosphate uses ribulose bisphosphate carboxylase (RUBISCO) b l C6 intermediate produced
most TP used to regenerate RuBP, but some used for carbohydrate, carbohydrate lipid and amino acid synthesis
Paradoxically, Paradoxically conditions that favour photosynthesis can actually reduce it!
Light intensity
as light intensity increases photosynthesis increases light compensation and saturation points are evident at light saturation point, another factor limits photosynthesis
Temperature
at low temperatures photosynthesis is limited, raising t i i temperature t increases rate to an optimum above optimum rate declines due to enzyme denaturation Note: at low light intensities temperature has no effect
photochemical processes are not affected by temperature
Investigating photosynthesis
p possible to measure photosynthesis by
CO2 uptake or O2 production
Accessory pigments
increase the range of g wavelengths of light that can be used
zonation of algae green brown red
must identify
independent variable, dependent variable and control variables
Some leaves are adapted for full f ll sun, others f shaded th for h d d conditions
sun leaves have higher LCP and LSP than shade leaves
Respiration
Select topic by clicking on the button General introduction Role of ATP Structure of ATP Stages of respiration Respiratory substrates p y Measurement of respiration Summary
Introduction ...
All living things need energy if they are to stay alive. It is needed for a range of living or biological processes such as: h
Active transport Mechanical work
muscle contraction
Anabolic reactions
synthesis complex molecules from simpler building blocks
As you are aware, the ultimate source of the energy used by most organisms is the sun. Plants harvest solar energy
convert it into chemical potential energy of photosynthetic products products.
Respiration converts the chemical potential energy of organic matter into chemical potential energy of ATP.
ATP is used by cells for work. It is the universal energy currency of the cell. Energy released by respiration is transferred to the highhigh-energy bond of ATP, which can then be used for work. k We will briefly look at the role of respiration (ATP) in
active t ti transport and t d metabolic reactions
Active transport
You are familiar with active transport from your AS and so are the experts! So What is active transport? The movement of materials against their concentration gradient What is needed for it to occur? A carrier and energy in the form of ATP
Many materials are moved against a concentration gradient: nutrients (digestive system) salt (loop of Henl in kidney) sodium and potassium (in nerve and other sensitive cells) to name but a f few.
SodiumSodium-potassium pump
Most cells have sodium-potassium pumps in their sodiumplasma membranes. They pump sodium ions out and potassium ions in. They are essential for creating the resting potential of nerve cells, without which action potentials/nerve ll ith t hi h ti t ti l / impulses could not be generated!
Role in metabolism
Cells need ATP energy to drive metabolism. Protein synthesis and DNA replication are anabolic reactions
build large complex molecules from simpler building blocks protein is built from amino acid monomers DNA is built from nucleotide monomers
nucleotides are a sugar, phosphate, base combination
Amino acids and nucleotides have to be activated with two extra phosphate groups before they can be built into a new protein or DNA.
A high-energy covalent bond is made between highthe carboxyl end of the amino acid and the 3 end of the tRNA makes an aminoacyl tRNA k i l tRNA.
AA AA + + P P P Adenosine + P Adenosine + P P
ATP
Aminoacyl tRNA
AMP
Pi
The DNA molecule has unzipped (hydrogen bonds holding complementary bases have been broken by a helicase enzyme) and the organic bases are exposed. A ti t d nucleotides ( d Activated l tid (nucleoside t i h l id triphosphates) b h t ) base pair with th i i ith their complementary bases (AT; GC) and the DNA polymerase removes the two phosphates from the activated nucleotides. This provides the energy to make the phosphodiester bond between the remaining phosphate and the growing DNA molecule.
ATP structure
Energy is not made directly available to cells
if all the energy contained within a glucose molecule were released all at once l d ll t it could not be used and much would be wasted
It is a phosphorylated nucleotide ATP is known as the cells energy currency it is used b all li i organisms t carry energy f d by ll living i to from
energyenergy-releasing reactions to energyenergy-consuming reactions
ATP structure: t t
adenine d i adenosine ribose
Once made, th ATP moves away from the site of O d the f th it f synthesis to the place where it is needed
ATP is broken down into ADP and inorganic phosphate releases the energy contained within the bond catalysed by an ATPase
The energy released is then used for work. The ADP and inorganic phosphate is recycled recycled.
Energy-producing reactions (respiration) allow energy to be transferred into the bond energy of ATP
31 kJ mol-1 31 kJ mol-1 14 kJ mol-1
ATP
ADP + P
AMP + P
Adenosine + P
31 kJ mol-1
31 kJ mol-1
14 kJ mol-1
Energy-consuming Energy consuming reactions (work e g active transport) use the energy (work, e.g. released from the bond energy of ATP. Realistically, it is only the first part of this sequence that is important for respiration, ATP production and use for work.
ATP
ADP +
ATP is made from ADP and inorganic phosphate using energy from energyenergy releasing reactions such as respiration. ATP synthetase catalyses this process.
ATP
ADP +
ATP is broken down into ADP and inorganic phosphate, phosphate releasing the energy contained within the bond. ATPase catalyses this process. The energy is then i th used f d for work. k
ATP
ADP +
Stages of respiration
Respiration is the process whereby organic molecules are broken down (oxidised) and the energy released used as f l f synthesis of ATP l d d fuel for th i f ATP. The main fuel is glucose, although other fuels can be used lipid protein Respiration/oxidation of energy rich organics like glucose can happen:
in the presence of oxygen in the absence of oxygen
Aerobic
in the presence of oxygen glucose is completely oxidised a lot of energy is released for the synthesis of a lot of ATP glucose + oxygen carbon dioxide + water + energy (2870 kJ)
Anaerobic A bi
in the absence of oxygen glucose i not f ll oxidised l is t fully idi d only a little energy is released, so much less ATP is produced glucose glucose ethanol + carbon dioxide + energy (150 kJ) lactate + energy (150 kJ)
Overall aerobic respiration can be summarised by an equation: word equation glucose + oxygen carbon dioxide + water + energy chemical equation C6H12O6 + 6O2 6CO2 + 6H2O + 2870 kJ
Glycolysis takes place in the cytoplasm and does not require O2 Link, Krebs and oxidative phosphorylation take place in the mitochondrion and do require O2
Coenzyme A
often abbreviated to CoA transfers acetate from the link reaction into Krebs cycle
Click to select topic on aspects of the stages of respiration Glycolysis Link reaction Krebs cycle Oxidative h O id ti phosphorylation h l ti Experimental evidence for chemiosmosis Mitochondrial structure and function Anaerobic respiration Energy release in respiration Efficiency of respiration
Glycolysis ...
Glycolysis is the first stage of respiration It takes place in the cytoplasm It is the first stage of both aerobic and anaerobic respiration and doesnt need O2 therefore it is doesn anaerobic (sometimes called anaerobic glycolysis) bi ( ti ll d bi l l i ) Essentially the stages of glycolysis split glucose (C6) to produce 2 molecules of pyruvate (C3) 2ATP (net) (C3), and 2 reduced NAD (sometimes referred to as NADH+ + H+ or NADH2)
Glycolysis includes a phosphorylation stage and an oxidation stage. In the phosphorylation stage
Glucose is phosphorylated by adding 2 phosphates
uses up 2 molecules of ATP l l f produces hexose bisphosphate
During the oxidation of triose phosphate enough energy is released to make 4x ATP
glycolysis
2ATP
Phosphorylation stage
2ADP
NAD
ADP ATP
NAD
Oxidation O id ti stage
NADred
Link reaction
Link reaction takes place in the mitochondrial matrix It connects glycolysis to Krebs cycle
happens when oxygen is available
link reaction
NAD
CO2 CoA
NAD red
CoA
Krebs cycle
Krebs cycle takes place in the mitochondrial matrix
it completes the oxidation of respiratory substrate
In K b I Krebs cycle l
the acetyl group is added to oxalo-acetate oxaloproduces citrate
Note: organic chemicals from other sources can feed into Krebs
acetyl groups from the oxidation of fatty acids organic acids following deamination of amino acids.
CoA
Krebs cycle
CoA
CO2
CO2
Oxidative phosphorylation
Oxidative phosphorylation also takes place in the mitochondrion It is the final stage of respiration
involves the transfer of hydrogen and then electrons along a transport chain chain. hydrogen is passed into the transport chain from
reduced NAD and reduced FAD.
When the l t Wh th electrons are transferred to O2, H+ i t f dt is removed from the mitochondrial solution to reduce it to water. water Note:
reducing power in reduced NAD (NADH2) and reduced FAD (FADH2) is used to generate ATP free oxygen is the terminal electron acceptor.
oxidative phosphorylation
Hydrogen transferred NADH2 FAD NAD FADH2 red ox ox cyt y red Electrons transferred between cytochromes red cyt ox ox cyt y red O2 H 2O ATP ATP ATP
The traditional story was that: that: as the protons, and then electrons are passed along the chain of hydrogen and electron carriers there is a loss of energy. the l th loss of energy is coupled t ATP synthesis f i l d to th i reduced NAD has sufficient energy for 3 ATPs reduced FAD has sufficient energy for 2 ATPs. ff f ATPs.
Chemiosmosis
The traditional view is not exactly what happens. It is true that:
reduced NAD and reduced FAD pass their hydrogen into the transport chain the h d th hydrogen splits i t H+ i lit into ions and electrons and th t d l t d that the electrons travel down the electron transport chain .
However, However it is now generally accepted that energy is used to pump protons from the matrix into the mitochondrial intermembrane space. p The protons then flow back into the matrix
via an ATP synthase y so generating ATP.
Note:
despite th name d it the
chemiosmosis has nothing to do with movement of water or water potential g p gradients
O2
Oxidative phosphorylation
ADP + Pi
H+
Krebs
H+ H+
ATP ADP + Pi
H+
ATP
ATP
H2O
Consistent with the idea that H+ ions are actively transported across the membrane
active because a gradient exists and is maintained.
alkaline acid
When illuminated the medium becomes alkaline and thylakoids become acid (pH 4)
same pH
When isolated chloroplasts are illuminated the pH of the th medium i which th di in hi h they are suspended becomes higher and the thylakoids become lower (pH 4). This observation is consistent with the idea that hydrogen ions are pumped into the thylakoids from th stroma. th l k id f the t
When not illuminated the medium and thylakoids remain the same pH
H+
H+
pH 8 buffer + ADP + Pi
pH 4 buffer + ADP + Pi
If isolated chloroplasts are suspended in an acid medium in the dark, the chloroplasts and thylakoids become acid. The chloroplast envelope can be disrupted and the thylakoids separated from the rest of the chloroplast debris. The thylakoids are then placed in buffered medium with ADP and Pi.
if the suspending medium is p g pH 8 ATP is synthesised. if the suspending medium is pH 4 ATP is not made made.
ATP is made
ATP is made by the thylakoids if there is a gradient between the thylakoid and the surrounding medium and ADP + Pi are present
mitochondria are the site of respiration they are surrounded by a double membrane or envelope the inner membrane is highly folded into cristae (singular crista) ( i l crista) i t there is a fluid filled region called the matrix
link reaction and Krebs cycle take place in the matrix which contains the th necessary enzymes electron transport or oxidative phosphorylation takes place in the inner membrane which b hi h contains the carriers and stalked particles or p ATP synthetase
TEM of a mammalian lung showing a mitochondrion. The double membrane or envelope, folded inner membrane making the cristae and matrix are visible. Public domain image: Louisa Howard.
Intermembrane space
Protons are pumped into the intermembrane space using the energy released by electron transport. The proton flow back into the matrix via an ATP synthetase, generating ATP.
Matrix
Contains the enzymes of link reaction and Krebs cycle. Hydrogen passes to ETC, CO2 excreted.
Crista
Foldings provide a large internal surface area. The membrane contains the electron transport chain and the stalked particles (ATP synthetase).
Envelope
TEM of a mammalian lung showing a mitochondrion. The double membrane or envelope, folded inner membrane making the cristae and matrix are visible. Public domain image: Louisa Howard.
Double membrane that surrounds the mitochondrion. Outer mitochondrion membrane is smooth, inner one is highly folded.
Inner membrane: Highly folded Less permeable Studded with 9nm diameter spheres the so-called sostalked particles Spheres are the ATP p synthetase
The inner membrane has the electron transport chain ETC pumps H+ ions into the intermembrane space, so creating a gradient between the intermembrane space and the matrix H+ ions flow back into the matrix through the stalked particles which synthesise ATP
Anaerobic respiration
What happens if oxygen is not available? The last stages cannot take place because the electrons cannot be passed onto oxygen. So
the electron acceptors cannot accept any more from reduced NAD and reduced FAD NAD and FAD cannot be regenerated meaning there is no NAD or FAD to accept hydrogen from the link reaction and Krebs cycle respiration in the mitochondrion stops.
anaerobic respiration
ATP
reduced NAD
NAD
Pyruvate
Lactate
The oxygen needed by the hepatocytes to remove the l ti th lactic acid i k id is known as th oxygen d bt. the debt. debt
Ethanol is toxic
eventually alcoholic f t ll l h li fermentation stops t ti t
The oxidation of glucose is incomplete and so much energy remains in the ethanol
can be a significant intake of energy for drinkers
anaerobic respiration
ATP
reduced NAD
NAD
CO2
Pyruvate
Ethanal
Ethanol
Products
Significance/other
Realistically, any hexose could be the substrate as they can be converted from one to another. Products Significance/other Lactic acid Pyruvate accepts the hydrogen, regenerating NAD so that glycolysis can continue. Build up is toxic. Causes oxygen debt debt. Ethanol Carbon dioxide Pyruvate converted to ethanal which then accepts the hydrogen regenerating hydrogen, NAD so that glycolysis can continue. Build up is toxic. toxic Ethanol contains a lot of energy calorific intake!
Complete the energy b l C l h balance sheet f aerobic and h for bi d anaerobic respiration
Stage
Glycolysis
Aerobic
Anaerobic
subtotal Link/Krebs
subtotal
subtotal
subtotal b l Total
Stage
Glycolysis
Aerobic
2 ATP (net) 2 NADH2
Anaerobic
2 ATP (net)
subtotal Link/Krebs
subtotal
subtotal
subtotal Total
28 (34) 32 (38)
subtotal total
0 2
Efficiency of respiration
It is possible to calculate the efficiency of aerobic and anaerobic respiration Assume: 1 mole of sugar contains 2870kJ of energy. Each high energ phosphate bond of ATP prod ced energy produced contains 31kJ per mole. All 2870kJ is released in aerobic respiration whilst respiration, only 150kJ is released under anaerobic conditions.
Efficiency can be calculated as: usable energy produced x 100/ energy released Where:
usable energy is that transferred to ATP energy released is the total energy released Calculate the efficiency of aerobic and anaerobic respiration
Aerobic respiration:
usable energy produced x 100 / energy released Best: es (38 x 31) x 100 / 2870 = 41.0% Realistic: (32 x 31) x 100 / 2870 = 34.6%
Anaerobic respiration:
usable energy produced x 100 / energy released Best/realistic: es / ea s c (2 x 31) x 100 / 150 = 41.3%
When you have performed the calculations you might be surprised to find that both aerobic and anaerobic respiration h i ti have about 40% efficiency, i 40% of th b t ffi i i.e. f the energy released is trapped in ATP** That said, only about 5% of the energy contained in said glucose is actually released under anaerobic co d t o s conditions what happens to the rest? The 60% or so is lost as heat energy! lost Energy still present in the lactate or ethanol!
**depends upon your calculated value for ATP produced!
Respiratory substrates
A respiratory substrate is an organic molecule that can be respired (oxidised) to produce usable energy in th form of ATP. i the f f ATP
carbohydrates (glucose) are important respiratory substrates
only one for some tissue e.g. brain cells and red blood cells.
glycerol
converted to pyruvate fed into Krebs cycle via the link reaction.
The relationship between the respiratory substrates and respiration is summarised in the following diagram. di Annotate your copy of it to indicate the reactions that occur. occur
glucose
triose phosphate reducing power ATP pyruvate CO2 glycerol lipid fatty acid
reducing power
Acetyl CoA
C4 acid
reducing power
ATP
So, from the following equation th t summarises S f th f ll i ti that i respiration C6H12O6 + 6O2 6CO2 + 6H2O + energy Which are the substrates? glucose/carbohydrate and O2 l / b h d t d Which are the products? CO2 and water d t
Traditionally, either O2 consumption or CO2 production would be used to measure respiration Suggest how each could be measured O2 consumption Oxygen electrode Use respirometer and CO2 scrubber to measure O2 used d CO2 production pH change
CO2 is an acid gas, add it and pH drops
A traditional way of measuring respiration is to measure the amount of oxygen used the more oxygen consumed in unit time, the faster the yg yg rate of respiration. There are several types of simple respirometer available, one type is illustrated in the diagram:
Manometer M t fluid/ink
Living organism (small invertebrate) in mesh wick CO2 scrubber e.g. NaOH
Respiration removes O2 and produces CO2 d d CO2 is removed by the scrubber The volume of gas in the chamber drops Manometer fluid moves M t fl id towards the chamber Movement proportional p p to O2 uptake
The distance moved is proportional to the rate of respiration, more specifically to the oxygen consumption What other information is needed to calculate the volume of oxygen consumed and the actual rate of oxygen consumption? internal diameter/radius of the capillary tube crosscross-sectional area of the capillary tube time What formula would you apply to measure the rate of oxygen consumption? p r2 x distance moved / time Do any factors need controlling so that we are confident in our estimate of respiration? Working in p p g pairs/small g p groups, describe and explain the effect of each factor and its control.
Factor Temperature
Effect if uncontrolled
Control measure
Atmospheric pressure
Other
Factor Temperature
Effect if uncontrolled Temperature influences respiration increases if temperature rises but does not exceed the optimum. p Thermal expansion of gas. Influences volume of gas and movement of the manometer fluid or ink high pressure moves ink towards the chamber, chamber low pressure away away. More/less CO2 scrubber in large/small respirometer affects efficiency of CO2 removal. More efficient with more scrubber and less volume Amount of respiring material Age of material Physiological t t Ph i l i l state
Control measure Water bath or room temperature, with frequent checks on temperature. Choose optimum. p Using a (de)compression chamber is not realistic! Use a control minus organism movement in control used to compensate... compensate Standardise!
Atmospheric pressure
Other
Glass beads
Living organism
NaOH
NaOH
Water bath
Fluctuations in temperature
cause fluctuation in respiration rate affect the volume of gas in the respirometer
and so the accuracy of measurement an increase in temperature causes thermal expansion p p cooling causes thermal contraction.
Summary
Plants, animals and microorganisms must respire
need ATP for
active transport ti t t
sodium potassium pump glucose
During respiration
substrates like glucose are b t t lik l oxidised
energy released used to produce ATP
metabolic reactions
DNA synthesis protein synthesis
Under aerobic conditions oxidation is complete Respiration divided into four stages:
glycolysis link reaction Krebs cycle and y oxidative phosphorylation
transfers energy
from energy producing reactions to energy consuming reactions
Respiration is
controlled b enzymes t ll d by involves coenzymes like NAD and acetyl coenzyme A
Glycolysis
takes place in the cytoplasm p y p starts with phosphorylation of glucose to hexose bisphosphate hexose bisphosphate splits to give 2x triose phosphate further oxidised to pyruvate yields
2x ATP (net) and 2x reduced NAD
decarboyxlated to produce acetate and reduced NAD acetate combined with coenzyme A to be carried to next stage
Krebs cycle
takes place in acetate (2C) added to oxaloacetate (4C) and produces citrate (6C) citrate is then reconverted to oxaloacetate
involves a series of
decarboxylations dehydrogenations and substrate level phosphorylation
Under aerobic conditions pyruvate is actively transported into the mitochondrial matrix Link reaction
pyruvate enters the link reaction
Oxidative phosphorylation
where most of the ATP made takes place in the inner membrane (cristae) (cristae) reducing power (h d d i (hydrogen) ) fed into oxidative phosphorylation from
reduced NAD and reduced FAD
hydrogen, hydrogen then electrons pass along carriers energy in the hydrogen and e- is used to produce ATP protons pumped into the intermembrane space flow back into the matrix via an ATP synthetase (stalked particle) chemiosomosis
matrix has
link and Krebs cycle enzymes small ribosomes and circular DNA
ethanol pathway
pyruvate converted to ethanal ethanal reduced to ethanol ethanol still contains much energy
Aerobic respiration
Under anaerobic conditions link, Krebs li k K b cycle and l d oxidative phosphorylation cannot continue Limited ATP production is possible if glycolysis continues NAD must be regenerated
lactate and ethanol pathway regenerate NAD t
lactate pathway
py pyruvate reduced to lactate note energy still present in lactate
Lipids are more highly reduced than amino acids, so produce more energy