Topic 8 Metabolism

8.1 Metabolism

8. Metabolic Metabolism is the web of all the enzyme-catalysed

1 pathways reactions in a cell or organism in order to maintain life
U1 consist of  Most chemical changes in a cell do not occur in one
chains and large jump, but result from a series of reactions known
cycles of as a metabolic pathway, with each step controlled by a
enzyme- specific enzyme
catalysed  A very simple, generalised metabolic pathway is
reactions. represented below

 Each arrow represents a specific enzyme that causes

one substrate to be changed to another, until the final
product of the pathway is formed
 Metabolic pathways are usually carried out in
designated compartments of the cell where the
necessary enzymes are clustered and isolated

Metabolic pathways are typically organised into chains or

cycles of enzyme-catalysed reactions
 Ex of chains: Glycolysis in cell respiration
 Ex of cycles: Krebs cycle in cell respiration and Calvin
cycle in photosynthesis

8. Enzymes lower Every chemical reaction requires a certain amount of

1 the activation energy in order to proceed
U2 energy of the  Substrates must pass through a transition state before
chemical they are converted into products
reactions that  Energy is required to reach the transition state – this is
they catalyse. activation energy (EA)
 Activation energy is used to break or weaken bonds in
the substrates

When an enzyme catalyses a reaction, the substrate bind to

the active site
 This binding stresses and destabilises bonds in the
substrate, reducing the overall energy level of the
substrate’s transitionary state  the activation energy
of the reaction is thus reduced
 The net energy change of the reaction is unaffected by
the involvement of the enzyme, but the rate of reaction
is greatly increased by the reduction in activation
energy
8. Enzyme Normal enzyme action
1 inhibitors can  In a normal reaction, a substrate binds to an enzyme via
U3 be competitive the active site to form an enzyme-substrate complex
or non-  The shape and properties of the substrate and active
competitive. site are complementary, resulting in enzyme-substrate
specificity
 When binding occurs, the active site undergoes a
conformational change to optimally interact with the
substrate (induced fit)
 This conformational change destabilises chemical
bonds within the substrate, lowering the activation
energy
 As a consequence of enzyme interaction, substrate is
converted into product at an accelerated rate

Enzyme inhibition
An enzyme inhibitor is a molecule that disrupts the normal
reaction pathway between an enzyme and a substrate by
preventing the formation of an enzyme-substrate complex
 Enzyme inhibitors can be either competitive or non-
competitive
 Inhibition of enzymes may be either reversible or
irreversible

A competitive inhibitor directly interferes with an enzyme’s

active site by binding to it so that the substrate cannot
bind
 Structurally and chemically similar to substrate
 Inhibitory effects can be reduced by increasing
substrate concentration
Ex of competitive:
 Relenza is a synthetic drug to treat individuals infected
by the influenza virus
 Viruses are released from infected cells when the viral
enzyme neuraminidase cleaves a docking protein
 Relenza competitively binds to the neuraminidase active
site and prevents cleavage of the docking protein
 Consequently, viruses are not released from infected
cells, preventing spread of influenza virus
 A non-competitive inhibitor binds to an enzyme at a
location other than the active site (allosteric site), causing
a conformational change in the enzyme so that the
substrate cannot bind
 As inhibitor is not in direct competition with the
substrate, increasing substrate levels cannot mitigate
the inhibitor’s effect
Ex of non-competitive:
 Cyanide is a poison which prevents ATP production via
aerobic respiration, leading to eventual death
 It binds to an allosteric site on cytochrome oxidase, a
carrier molecule that forms part of the electron
transport chain
 By changing the shape of the active site, the electron
transport chain cannot continue to function, and ATP is
not produced via aerobic respiration

8. Distinguish
1 different types
S1 of inhibition
from graphs at
specified
substrate
concentration.

8. Metabolic End-product inhibition is a form of negative feedback by

1 pathways can which metabolic pathways can be controlled
U4 be controlled  In end-product inhibition, the final product in a series of
by end-product reactions inhibits an enzyme from an earlier step in the
inhibition. sequence
 The product binds to an allosteric site and temporarily
inactivates the enzyme via non-competitive inhibition
 As the enzyme can no longer function, the reaction
sequence is halted, and the rate of product formation is
decreased

End-product inhibition functions to ensure levels of an

essential product are always tightly regulated
 If product levels build up, the product inhibits the
reaction pathway and hence decreases the rate of
further product formation
 If product levels drop, the reaction pathway will proceed
unhindered and the rate of product formation will
increase
8. End-product Isoleucine is an essential amino
1 inhibition of acid, meaning it is not synthesised
A1 the pathway by the body in humans. Plants and
that converts bacteria synthesise isoleucine from
threonine to threonine in a five-step reaction
isoleucine. pathway.
 As concentration of isoleucine
builds up, it binds to the
allosteric site on the enzyme in
the first step of the chain,
threonine deaminase
 This acts as a non-competitive
end-product inhibitor, and the
rate of isoleucine formation is decreased

8. Use of Malaria is a disease caused by pathogen Plasmodium

1 databases to falciparum
A2 identify  The increasing resistance of P. falciparum to anti-
potential new malarial drugs such as chloroquine drives the need to
anti-malarial develop new anti-malarial drugs
drugs.  The maturation and development of the parasite in both
human and mosquito hosts is coordinated by specific
enzymes
 By targeting these enzymes for inhibition, new anti-
malarial drugs and medications can be produced

Scientists have sequenced the genome of infectious

species of plasmodium and used it to determine the
parasite’s proteome (entire set of proteins expressed by a
genome)
 From the proteome, enzymes involved in parasitic
metabolism have been identified as potential targets for
drug inhibition

These enzymes may be screened against a bioinformatic

database of chemicals to identify potential enzyme
inhibitors
 Once a promising compound is identified, it may be
chemically modified to improve its binding affinity and
lower its toxicity
 In one study, over 300,000 chemicals were screened to
identify 19 new chemicals that might function as
inhibitors
8. Calculating The rate of an enzyme-catalysed reaction can be calculated
1 and plotting and plotted according to the time taken for the reaction to
S2 rates of proceed
reaction from  The time taken can be measured according to either the
raw amount of product formed or amount of substrate
experimental consumed
results.  Reaction rate is the inverse of time taken, meaning that
the reaction rate is higher when less time is taken
1
 Formula for rate of reaction: Rate=
timetaken ( s )
 Factors which can influence the rate of an enzyme-
catalysed reaction include temperature, pH, and
substrate concentration
 8.2 Cellular respiration

8.2 Cellular Metabolism is the web of all the enzyme-catalysed

U1 respiration reactions in a cell or organism in order to maintain life.
involves the The reactions involve:
oxidation and  Catabolic pathways, which result in the breakdown of
reduction of complex molecules into smaller molecules. Ex: cellular
electron respiration
carriers.  Anabolic pathways, which result in the synthesis of
more complex molecules from simpler molecules. Ex:
photosynthesis

Oxidation and reduction can be compared as follows

(OILRIG):

Electron carriers are substances that accept and give up

electrons are required. They often link oxidation and
reduction reactions in cells.
 The main electron carrier in cellular respiration is NAD.
In photosynthesis, a phosphorylated version of NAD is
used – NADP

Reduction of coenzyme NAD

NAD initially has a positive charge and exists as NAD+. It
accepts two electrons in the following way:
 Two hydrogen atoms are removed from the substance
that is being oxidised (oxidation = loss of hydrogen)
 One of the hydrogen atoms is split into a proton and an
electron
 NAD+ accepts the electron and the proton (H+) is
released  NAD
 NAD accepts both the electron and proton of the other

hydrogen atom
This reaction demonstrates that reduction can be achieved
by accepting atoms of hydrogen, because they have an
electron. Oxidation can therefore be achieved by losing
hydrogen atoms.

8.2 Phosphorylatio Phosphorylation is the addition of a phosphate molecule

U2 n of molecules (PO43-) to an organic molecule
 makes them  For many reactions, the purpose of phosphorylation is
less stable. to make the phosphorylated molecule more unstable;
i.e. more likely to react

Adenosine triphosphate (ATP) is a high-energy molecule

that functions as an immediate power source for cells
 One molecule of ATP contains three covalently bonded
phosphate groups – which store potential energy in
their bonds
 Phosphorylation makes the molecule less stable, and
hence ATP is readily reactive
 When ATP is hydrolysed to form ADP + Pi, the energy
stored in the terminal phosphate bond is released for
use by the cell

Two functions of ATP:

1. Provides energy for cell metabolism
2. Phosphorylates other molecules by releasing
phosphate group  rendering them more reactive
Ex of a metabolic process that uses ATP: first reaction in
glycolysis

 The conversion of glucose to glucose-6-phosphate

absorbs energy and the hydrolysis of ATP releases
energy
 Because the reactions are coupled, they proceed
simultaneously

8.2 In glycolysis, Glycolysis (cytosol) is the controlled breakdown of

U3 glucose is carbohydrates, occurs in the cytosol, and uses no oxygen
converted to  In glycolysis, a hexose sugar (6C) is broken down into
pyruvate. two molecules of pyruvate (3C)

8.2 Glycolysis Stages of glycolysis (substrate-level phosphorylation, lysis,

U4 gives a small oxidation, and ATP formation)
net gain of  A hexose sugar (typically glucose) is phosphorylated by
ATP without two molecules of ATP to form a hexose biphosphate.
the use of
oxygen.

Net gain of
ATP: 2 –
produces 4 but
uses 2 to
initiate

Phosphorylation makes the hexose sugar less stable

and more reactive
  The less stable hexose biphosphate is lysed into two 3-
carbon sugars (lysis)
 Two hydrogen atoms are removed from each of the 3C
sugars to reduce NAD+ to NADH (+H+). Two molecules of
NADH are produced in total (one from each 3C). As
NADH is formed, released energy is used to add an
inorganic phosphate to each 3C compound (oxidation)
 Enzymes then remove the phosphate groups so that
they can be added to ADP to synthesise ADP, using

energy released by the reduction of NAD+ (ATP

formation)
The end result is the formation of four molecules of ATP
(two molecules used to initiate glycolysis so net ATP = 2),
two molecules of NADH, and two molecules pf pyruvate, in
the ionised form of pyruvic acid

8.2 In aerobic cell Glycolysis occurs in the cytoplasm and produces small
U5 respiration, amounts of ATP and a product known as pyruvate
pyruvate is  If no oxygen is available, the pyruvate enters into
decarboxylate anaerobic respiration, which occurs in the cytoplasm
d and oxidised. and does not result in any further production of ATP
 The products of anaerobic respiration are lactate or
ethanol and carbon dioxide

If oxygen is available, the pyruvates undergo aerobic

respiration in the mitochondria of the cell
 It is decarboxylated and oxidised in the mitochondrion,
resulting in the production of a large number of ATPs,
carbon dioxide, and water

This occurs in several stages: Link reaction, Krebs cycle,

Oxidative phosphorylation (electron transport chain &
chemiosmosis)

8.2 In the link Link reaction

U6 reaction, Pyruvate enters the matrix of the mitochondria via active
pyruvate is transport
converted into 1. Pyruvate is decarboxylated, a reaction involving the
acetyl loss of a carbon in the form of carbon dioxide, to form
coenzyme A. the 2C acetyl group. CO2 is released as waste.
2. Acetyl group is then oxidised with the reduction of
Net gain of NAD+ to form NADH (+H+)
ATP: 0 3. Acetyl group combines with coenzyme A to form acetyl

coenzyme A (acetyl CoA)

 If ATP levels in the cell are high, acetyl CoA can be
synthesised into a lipid for storage
 If cellular ATP levels are low, acetyl CoA enters the
Krebs cycle

8.2 In the Krebs The Krebs cycle (matrix) occurs in the matrix of the
U7 cycle, the mitochondrion and is referred to as a cycle as it begins
oxidation of and ends with the same substance.
acetyl groups 1. The 2C acetyl CoA from the link reaction combines with
is coupled to a 4C compound to produce a 6C compound
the reduction 2. The 6C compound is oxidised and decarboxylated by
of hydrogen the reduction of NAD+ to form a 5C compound. The
carriers, carbon is combined with oxygen and released from the
liberating cell as carbon dioxide.
carbon 3. The 5C compound is oxidised and decarboxylated to
dioxide. form a 4C compound, with another NAD+ reduced to
form NADH. Again, removed carbon is combined with
Net gain of oxygen and released as CO2.
ATP: 2 4. The 4C compound undergoes various changes involving
several processes, including reduction of a NAD+ to
form another NADH, reduction of FAD to form FADH2,
and phosphorylation of ADP to form ATP. The 4C
compound is changed during these steps to reform the
starting 4C compound of the cycle, allowing for the
cycle to restart.
 For each molecule of glucose, the Krebs cycle will run
twice, as:
1 glucose  2 pyruvates  2 acetyl CoA  cycle runs
twice

Products from one glucose molecule resulting from two

Krebs cycles:
 2 ATP
 4 CO2
 6 NADH (which allow energy storage and transfer)
 2 FADH2

State of ATP  Net gain of 4 ATPs – while six are generated (four from
synthesis after glycolysis and two from the Krebs cycle), two are used
glycolysis and to start glycolysis
Krebs cycle  Each of these ATPs have been produced by substrate-
level phosphorylation.
 Ultimately, the breakdown of each glucose molecule
results in a net gain of 36 ATPs
 The electron transport chain is where most of the ATPs
from glucose catabolism are produced
8.2 Energy released In aerobic respiration, there are several points where
U8 by oxidation energy released by oxidation reactions is coupled to the
reactions is reduction of mainly NAD, but also FAD
carried to the
 Reduced NAD (NADH) is produced during glycolysis, the
cristae of inner
link reaction, and Krebs cycle
mitochondrial
membrane by  FADH2 is produced during the Krebs cycle
reduced NAD
and FAD. Final part of aerobic respiration is called oxidative
phosphorylation, because ADP is phosphorylated to
“Energy” as in produce ATP, using energy released by oxidation
reducing power.  Substances oxidised include the FADH2 generated in
FADH2 and the Krebs cycle and the reduced NAD (NADH)
NADH can be
generated in glycolysis, the link reaction, and the Krebs
oxidised,
cycle
allowing for the
reduction of  Thus, it can be said that these molecules “carry” the
another energy released in pre-oxidative phosphorylation stages
molecule. to the mitochondrial cristae

8.2 Transfer of Oxidative phosphorylation (32 ATPs per glucose molecule)

U9 electrons Oxidative phosphorylation synthesises ATP using energy
between derived from oxidation of reduced hydrogen carriers (NADH
carriers in the & FADH2) from the previous stages of cell respiration
electron (glycolysis, link reaction, Krebs cycle)
transport  Oxidative phosphorylation involves the electron
chain is transport chain and chemiosmosis
coupled to  Where most ATPs are produced and first stage where
proton oxygen is needed
pumping.  Occurs within mitochondrion, but unlike the Krebs
cycle which occurred in the matrix, oxidative
8.2 In phosphorylation occurs on the inner mitochondrial
U1 chemiosmosis membrane
0 protons diffuse  Inner membrane is arranged into cristae (folds), which
through ATP increases surface area available for oxidative
synthase to phosphorylation
generate ATP.

8.2 Oxygen is Oxidative phosphorylation occurs over a number of distinct

U1 needed to bind steps:
1 with the free 1. Proton pumps create an electrochemical gradient
protons to (proton motive force) – electron transport chain
form water to 2. ATP synthase uses the subsequent diffusion of protons
maintain the (chemiosmosis) to synthesise ATP
hydrogen 3. Oxygen accepts electrons and protons to form water
gradient.
Step 1: Generating a proton motive force via electron
Net gain of transport chain
ATP: 32  Hydrogen carriers (NADH and FADH2) are oxidised and
releases high energy electrons and protons (H+)
 These high-energy electrons are transferred to the
electron transport chain, which consists of several
membrane carrier proteins
 NADH + H+ supplies pairs of hydrogen atoms to the first
carrier in the chain, with NAD+ (after oxidation)
 returning back to the matrix
 The two hydrogen atoms are split to release two
electrons, which pass from carrier to carrier in the
chain
 Energy is released as electron passes from carrier to
carrier, and this energy is used to transfer protons (H+)
across the inner mitochondrial membrane into the
intermembrane space (from the matrix)
 As electrons continue to flow along the chain and more
protons are pumped across into the inner mitochondrial
membrane, an electrochemical gradient builds up (high
[H+] in intermembrane space, low in matrix), storing
potential energy (proton motive force)

Step 2: ATP synthesis via chemiosmosis

 Proton motive force will cause H+ ions to move down
their electrochemical gradient and diffuse back into the
matrix
 Diffusion of protons is called chemiosmosis and is
facilitated by the transmembrane enzyme ATP synthase
 As the H+ ions move through ATP synthase they trigger
molecular rotation of the enzyme, synthesising ATP by
phosphorylation of ADP

Step 3: Reduction of oxygen

 To allow electrons to
continue to flow, they
must be transferred to a
terminal electron
acceptor at the end of
the chain – in aerobic
respiration, this is
oxygen
 Oxygen briefly becomes
•O2-, but then combines
with two H+ ions from
the matrix to become
water
 In the absence of
oxygen, hydrogen carriers cannot continue to transfer
 energised electrons to the chain and ATP production is
halted (anaerobic respiration)

Summary of ATP We have now described the complete catabolism of one

production in molecule of glucose.
aerobic cellular  Raw materials were glucose and oxygen, but many
respiration enzymes, carriers, other molecules are also involved
 Products are carbon dioxide, water, and ATP
Glycolysis  link
reaction  Krebs
cycle  electron
transport chain

8.2 Analysis of Types of aerobic reactions

S1 diagrams of  Aerobic respiration involves three main types of
the pathways reactions – decarboxylation, oxidation, and
of aerobic phosphorylation
respiration to
deduce where
decarboxylatio
n and
oxidation
reactions
occurs.
Decarboxylation:
 Carbon atoms are removed from the organic molecule
(glucose) to form carbon dioxide
 Aerobic respiration involves the complete combustion
of glucose (6C) – so 6 CO2 molecules are produced

Oxidation:
 Electrons and hydrogen ions are removed from glucose
and taken up by hydrogen carriers (NAD+  NADH, FAD
 FADH2)
 Hydrogen carriers are oxidised in the electron transport
chain, where the energy released is used to make ATP
 Electrons and hydrogen ions are then taken up by
oxygen (reduction) to form water molecules
 12 hydrogen carriers are produced so 6 oxygen
molecules (O2) are required (12 × O = 6 × O2)

Phosphorylation
 Phosphorylation of hexose sugar at the beginning of
glycolysis
 Energy released from the breakdown of glucose is used
to phosphorylate ADP to make ATP
 Net total of 4 ATP molecules are produced directly via
substrate level phosphorylation during glycolysis (4 – 2)
 and Krebs cycle (2)
 Remaining 32 ATPs are produced indirectly via
oxidative phosphorylation via the electron transport
chain

 NADH molecules in the matrix (from Krebs cycle)

donate electrons to the start of the chain and produce 3
ATP each
 NADH molecules from the cytosol (from glycolysis)
produce 2 ATP each as they donate electrons later in
the chain
 FADH2 also only produce 2 ATP each for the same
reason

8.2 The structure Mitochondria are the powerhouses of the cell –

U1 of the synthesising large amounts of ATP via aerobic respiration
2 mitochondrion  All eukaryotic cells possess mitochondria, this is where
is adapted to oxidative phosphorylation occurs
the function it  Aerobic prokaryotes use the cell membrane to perform
performs. oxidative phosphorylation

Structure of the mitochondrion is adapted to the function it

performs:
 Outer membrane: outer membrane contains transport
proteins that enable the transport of pyruvate from the
cytosol
 Inner membrane: contains the electron transport chain
and ATP synthase (used for oxidative phosphorylation)
 Cristae: folds of the inner membrane, increasing the
SA:Vol ratio  more available for surface for oxidative
phosphorylation
 Intermembrane space: small space between
membranes that accumulates protons to create a
proton gradient for oxidative phosphorylation
 Matrix: central cavity that contains appropriate
enzymes and a suitable pH for the Krebs cycle to occur
8.2 Annotation of
S2 a diagram to
indicate the
adaptations of
a
mitochondrion
to its
functions.

8.2 Electron Electron tomography is a technique by which the 3-

A1 tomography dimensional internal structure of a sample can be
used to modelled
produce  Samples are repeatedly imaged using a transmission
images of electron microscope
active  Following each image, the sample is tilted to a different
mitochondria. angle relative to the electron beam
 Images are then compiled and used to computationally
reconstruct a 3D representation, called a tomogram
 8.3 Photosynthesis

8.3 Light- Photosynthesis uses light energy, carbon dioxide and water
U1 dependent
reactions
take place in
the to form glucose, water, and oxygen. The overall equation is:
intermembran  Water occurs on both sides because 12 molecules are
e space of consumed and 6 are produced
the  Photosynthesis is essentially the reverse of respiration,
thylakoids. and is an anabolic process
 Occurs in organisms known as autotrophs. Non-
8.3 Reduced
photosynthetic and non-chemosynthetic organisms are
U2 NADP and
referred to as heterotrophs. They must obtain their food
ATP are
from other organisms
produced in
the light-
Photosynthesis occurs in two major stages:
dependent
 Light-dependent reaction
reactions.
 Light-independent reaction
8.3 Light-
U3 independent The light-dependent reaction
reactions The light-dependent reaction occurs in the thylakoids or
take place in grana of the chloroplast. A stack of thylakoids makes up a
the stroma. granum (pl. grana).
 Light supplies energy for this reaction to occur –
8.3 Absorption of although the ultimate source of light is the sun, plants
U4 light by may survive well on light sources other than the Sun
photosystems
generates To absorb light, plants have special molecules called
excited pigments. There are several different pigments in plants,
electrons. and each effectively absorb photons of light at different
wavelengths
8.3 Photolysis of
 The two major pigment groups are chlorophylls and
U5 water
carotenoids
generates
 These pigments are organised on the membranes of the
electrons for
thylakoids in regions known as photosystems
use in the
light-
Photosystems are the light-absorbing regions on thylakoids
dependent
are include:
reactions.
 Chlorophyll a molecules
8.3 Transfer of  Accessory pigments
U6 excited  A protein matrix
electrons
occurs The reaction centre is the portion of the photosystem that
between contains:
carriers in  A pair of special chlorophyll a molecules
thylakoid  A protein matrix
membranes.  A primary electron acceptor

8.3 Excited While photosynthetic bacteria only have one type of

U7 electrons
 from photosystem, modern-day plants have two types of
Photosystem photosystem. Each absorbs light most efficiently at a
II are used to different wavelength.
generate a  Photosystem I is most efficient at 700 nm – P700
proton  Photosystem II is most efficient at 680 nm – P680
gradient.
Overall, the two photosystems work together to bring about
8.3 ATP synthase a non-cyclical electron transfer. This occurs in several
U8 in thylakoids steps:
generates 1. A photon of light is absorbed by a pigment in
ATP using the Photosystem II and is transferred to other pigment
proton molecules until it reaches one of the chlorophyll a (P680)
gradient. molecules in the reaction centre.
8.3 Excited 2. The photon energy excites one of the chlorophyll a
U9 electrons electrons to a higher energy state. This energised
from electron is captured by the primary electron acceptor of
Photosystem the reaction centre.
I are used to 3. The excited electron from step 2 pass from the primary
reduce NADP. acceptor down an electron transport chain, losing
energy at each of the three electron carriers. The first
8.3 In the light- electron carrier is plastoquinone (PQ) and the middle
U1 independent carrier is a cytochrome complex.
0 reactions a 4. Energy lost from electrons moving down the electron
carboxylase transport chain drives chemiosmosis to bring about
catalyses the phosphorylation of ADP to produce ATP.
carboxylation 5. A photon of light is absorbed by a pigment in
of ribulose Photosystem I. This energy is transferred through
biphosphate. several accessory pigments until received by a
chlorophyll a molecule (P700). Similar to photosystem II,
8.3 Glycerate 3- this results in an electron with a higher energy state
U1 phosphate is being transferred to the primary electron acceptor. The
1 reduced to de-energised electron from Photosystem II replaces the
triose lost electron on the chlorophyll a molecule.
phosphate 6. After being accepted by the primary electron acceptor
using reduced at the reaction centre of Photosystem I, the excited
NADP and electron is passed down a second electron transport
ATP. chain that only involves one electron carrier –
ferredoxin.
8.3 Triose
7. The enzyme NADP reductase catalyses the transfer of
U1 phosphate is
the electron from ferredoxin to the electron carrier
2 used to
NADP+. Two electrons are required to reduce NADP+ fully
regenerate
to NADPH.
RuBP and
8. Water is lysed by an enzyme to produce electrons,
produce
hydrogen ions, and an oxygen atom. This process is
carbohydrate
driven by light energy and is called photolysis. The
s.
electrons are supplied one by one to the chlorophyll a
8.3 Ribulose molecules of the reaction centre. This is to supply
U1 biphosphate electrons for the continuation of the light-dependent
3 is reformed reaction.
using ATP.
 The light-
dependent

NADPH and ATP are the final products of the light-

dependent reaction
 These two products supply energy for the light-
independent reaction to occur
 Photolysis of water also shows the origin of the oxygen
released by photosynthetic plants

Note that step 4 mentions chemiosmosis (phosphorylation

of ADP to produce ATP) in photosynthesis 
photophosphorylation
 In photosynthesis, chemiosmosis occurs between
Photosystem II and Photosystem I
 The cytochrome complex (b6-f) pumps hydrogen ions
from the stroma (cytosol-like region of chloroplast) into
the thylakoid space
 This increases concentration of these ions, which then
passively move through the ATP synthase protein
channel, the proton motive force of which then provides
energy to phosphorylate ADP

Below is a comparison of chemiosmosis in respiration and

photosynthesis:
 The light-independent reaction
The light-independent reaction occurs within the stroma
(cytosol-like region in the chloroplast)
 The products of the light-dependent reaction (ATP &
NADPH) drives the light-independent reaction: ATP
provides energy and NADPH provides reducing power
*Names of all (can bring about reduction of another molecule by
listed oxidising itself)
compounds  Light-independent reaction responsible for production of
are important glucose

The light independent reaction involves the Calvin cycle,

which occurs in the stroma of the chloroplast. It begins and
ends with the same substance.
1. Ribulose biphosphate (RuBP), a 5C compound, binds to
an incoming carbon dioxide (CO 2) in a process called
carbon fixation, which is catalysed by an enzyme called
rubisco. The result is an unstable 6C compound.
2. The unstable 6C compound breaks down into two 3C
compounds called glycerate 3-phosphate
3. The 3C molecules are reduced by ATP and the oxidation
of NADPH to form other 3C molecules called triose
phosphate.
4. Triose phosphate molecules then have two pathways:
some leave the cycle to become sugar phosphates that
may become glucose and complex carbohydrates
(starch & cellulose). Most, however, continue in the
cycle to reproduce the originating compound of the
RuBP (step 5)
5. Triose phosphate molecules produce RuBP molecules
using ATP
 The Calvin cycle
(part of light-
independent

Coefficients in front of each compound involved shows

what is required to produce one molecule of a 6-carbon
sugar (glucose)
 12 triose phosphate molecules are required to produce 1
molecule of glucose or 6 molecules of RuBP
 18 ATP and 12 NADPH molecules are required to
produce 6 RuBP or 1 molecule of glucose

Triose phosphate is the pivotal compound in the Calvin

cycle:
 It may be used to produce simple sugars such as
glucose, disaccharides such as sucrose, or
polysaccharides such as cellulose or starch
 However, most of it is used to regain the starting
compound of the Calvin cycle, ribulose biphosphate
(RuBP)

Summary of photosynthesis
 Process of photosynthesis involves the light-dependent
and light-independent reactions
*Note:  Products of light-dependent reaction are ATP and
ATP  ADP + NADPH, which are needed for the light-independent
Pi is a reaction to proceed
reduction
reaction. This
due to the
decrease in
oxidation
 state.

 Thus, in a way, the light-independent reaction also

requires light to occur, but not directly
 NADP+ and ATP move back and forth in the chloroplast
from the thylakoids to the stroma in their reduced
(NADPH and ADP) and oxidised (NADP+ and ATP) forms

Final summary of light-dependent and light-independent

reactions:

8.3 Calvin’s The light independent reactions are collectively known as

A1 experiment to the Calvin cycle
elucidate the  Calvin’s elucidation of photosynthetic carbon
carboxylation compounds is commonly classed the ‘lollipop
of RuBP. experiment’

Lollipop Experiment
 Radioactive carbon-14 is added to a ‘lollipop’ apparatus
containing green algae
 Light is shone on the apparatus to induce
photosynthesis of the algae, which will incorporate
carbon-14 into organic compounds through the Calvin
cycle
 After different periods of time, the algae is killed by
running it into a solution of heated alcohol, stopping cell
metabolism
 Dead algal samples are analysed using 2D
chromatography, which separates out the different
carbon compounds
 Any radioactive carbon compounds on the
chromatogram were then identified using
autoradiography
 By comparing different periods of light exposure, the
order by which carbon compounds are generated was
determined
 Calvin used this information to propose a sequence of
events known as the Calvin cycle
8. The structure The structure of the chloroplast allows the light-dependent
3 of the and light-independent reactions
U chloroplast is  Extensive membrane surface area of thylakoids – allows
1 adapted to its greater absorption of light by photosystems
function in  Small space (lumen) within thylakoids – allows for
photosynthes quicker achievement of a proton concentration gradient
is.  Stroma region similar to the cytosol of the cell – allows
an area for the enzymes necessary for the Calvin cycle to
8. Annotations work in
3 of a diagram  Double membraned organelle – isolates the working parts
S1 to indicate and enzymes of the chloroplast from the cytosol of the
the cell
adaptations
of a
chloroplast to
its function.