1. To know the importance of chemical energy in biological processes 2. To understand the role of ATP 3. To draw the structure of ATP 4. To understand the stages in aerobic respiration: glycolysis, link reaction, Kreb’s cycle and the electron transport chain 1. Movement e.g. movement of cilia and flagella, muscle contraction 2. Maintaining a constant body temperature to provide optimum internal environment for enzymes to function 3. Active transport – to move molecules and ions across the cell surface membrane against a concentration gradient 4. Anabolic processes e.g. synthesis of polysaccharides from sugars and proteins from amino acids 5. Bioluminescence – converting chemical energy into light e.g. ‘glow worms’ 6. Secretion – the packaging and transport of secretory products into vesicles in cells e.g. in the pancreas In pairs: Draw this grid on one Miniw’board. Put or on different sides of a second mwb (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) (15) (16) (1) Active transport uses carrier proteins (5) Phagocytosis is a type of endocytosis (9) Simple diffusion uses ATP (13) Endocytosis involves bulk transport out of a cell (2) Active transport needs ATP (3) Osmosis occurs from lower to higher water potential (7) Active (6) Facilitated transport occurs diffusion needs from lower to ATP (14) Facilitated higher conc. uses (10) diffusion Endocytosis channelbulk proteins (11) Pinocytosis involves is a type of transport exocytosis into a cell (14) Facilitated diffusion uses channel proteins (15) Diffusion occurs up a concentration gradient (4) Passive transport methods use ATP (8) Facilitated diffusion uses carrier proteins (12) Diffusion stops when equilibrium is reached (16) Exocytosis uses ATP Why is energy needed within cells? Allows chemical reactions to take place BUILD UP (synthesis) or BREAKDOWN of molecules In order to do this, energy is required to make and break bonds Where does the energy come from? The SUN is the ultimate source of energy for nearly all living organisms (the exceptions being a few deep sea chemosynthetic bacteria) Autotrophs make their own food (organic compounds) using carbon dioxide Heterotrophs assimilate energy by consuming plants or other animals Autotroph Organisms that can synthesise complex organic molecules from simple ones. There are two types of autotroph, depending on how they obtain their energy: i. Phototrophs: Autotrophs that use light energy e.g. Plants. ii. Chemotrophs: Autotrophs that use inorganic chemical energy e.g. sulphur bacteria ATP What provides the energy within cells? ATP…Adenosine Tri Phosphate Common to ALL living things Any chemical that interferes with the production or breakdown of ATP is fatal to the cell and therefore the organism •Chemical energy is stored in the phosphate bonds The role of ATP (adenosine triphosphate) The short term energy store of the cell Often called the ‘energy currency’ of the cell because it picks up energy from food in respiration and passes it on to power cell processes. ATP made up of: Adenine (a base) Ribose (a pentose sugar) 3 phosphate groups ATP Structure ATP is a nucleotide made from: 1. The nitrogenous base Adenine 2. A pentose sugar Ribose 3. Phosphate groups ATP: Function 1. It is a coenzyme involved in many enzyme reactions in cells. 2. It is the major energy currency of cells entrapping or releasing energy in most metabolic pathways. 3. The energy is released from ATP in a single step and in a small manageable amount. 4. It is a small molecule so will diffuse rapidly around the cell to where it is needed. 5. It is one of the monomers used in the synthesis of RNA and, after conversion to deoxyATP (dATP), DNA. ATP: and energy When the third phosphate group of ATP is removed by hydrolysis, a substantial amount of free energy is released, the exact amount depends on the conditions. For this reason, this bond is known as a "high-energy" bond. The bond between the first and second phosphates is also "high-energy". But note that the term is not being used in the same sense as the term "bond energy". In fact, these bonds are actually weak bonds with low bond energies. ATP + H2O -> ADP + Pi ADP is adenosine diphosphate. Pi is inorganic phosphate. * Hydrolysis: Decomposition of a substance by the insertion of water molecules between certain of its bonds. Food is digested by hydrolysis) *Free energy: The energy that can be harnessed to do work. How does ATP provide the energy? Chemical energy is stored in the phosphate bonds, particularly the last one To release the energy, a HYDROLYSIS reaction takes place to break the bond between the last two phosphate molecules Catalysed by ATP-ase ATP is broken down into ADP and Pi For each mole of ATP hydrolysed, about 34kJ of energy is released Some is lost, but the rest is useful and is used in cell reactions How ATP releases energy The 3 phosphate groups are joined together by 2 high energy bonds ATP can be hydrolysed to break a bond which releases a large amount of energy Hydrolysis of ATP to ADP (adenosine diphosphate) is catalysed by the enzyme ATPase (ATPase) ADP + Pi + 30 KJ mol-1 ATP (H2O) The 2nd phosphate group can also be removed by breaking another high energy bond. The hydrolysis of ADP to AMP (adenosine monophosphate) releases a similar amount of energy (ATPase) AMP + Pi + 30 KJ mol-1 ADP (H2O) AMP and ADP can be converted back to ATP by the addition of phosphate molecules The production of ATP – by phosphorylation - Adding phosphate molecules to ADP and AMP to produce ATP Phosphorylation is an endergonic reaction – energy is used Hydrolysis of ATP is exergonic - energy is released Advantages of ATP Instant source of energy in the cell Releases energy in small amounts as needed It is mobile and transports chemical energy to where it is needed IN the cell Universal energy carrier and can be used in many different chemical reactions What does this have to do with photosynthesis? ATP is both synthesised and broken down during photosynthesis! 6CO2 + 6H2O = C6H12O6 + 6O2 Light energy is required Chlorophyll Stored within chloroplasts 10-50 chloroplasts per plant cell An introduction… The Leaf Plant leaves areofflattened to The exchange gases through maximise theissurface areabyfor the the stomata regulated the absorption light. lie The guard cellsofwhich onupper eitherand side lower surfaces are covered bycells a of it. The palisade mesophyll waxy cuticle which the loss of are elongated and slows contain many water from thethis leaf. chloroplasts, is Beneath the mainthe cuticle lies the epidermis which photosynthetic area of the plant. provides some support for leaf. The spongy mesophyll hasthe large The lower epidermis has air spaces to allow for thesmall rapid pores called stomainthat forthe diffusion of gases andallow out of gaseous During the day leaf. Theexchange. veins in the leaf contain CO2 diffuses in and out,and during vascular tissue, the O2 xylem the night CO2 diffuses out and O2 phloem. The xylem provides in. Wateras vapour also escapes from support well as carrying water the it is this loss that andstomata mineraland nutrients. The phloem creates transpiration stream carries the away the products of drawing mineral nutrients the photosynthesis, primarily from sucrose, soil andrest up of into the plant. to the the plant. The Leaf Upper epidermis Palsade mesophyll Vein Vascular bundle Spongy mesophyll Lower Epidermis The Leaf Cuticle Upper epidermis Chloroplasts Palisade mesophyll Air space In spongy mesophyll Guard Cells and Stomata Stoma Guard Cells Lower epidermis Wynwood Reade, Martyrdom of Man, 1924 Photosynthesis – what we know (or should know!!...) “Building from light” Converts carbon dioxide into organic compounds Carried out by autotrophs All life either depends on it directly as a source of energy, or indirectly as the ultimate source of the energy in their food 6CO2 + 6H2O = C6H12O6 + 6O2 So how do we know all this?... The story starts a long time ago… Aristotle (384-322BC) Greek philosopher He proposed that plants, like animals, require food He concluded that green plants obtained their nourishment from the soil Aristotle’s theory was widely accepted until the 1600’s… Nicholas of Cusa (1401-1464) Cardinal of the Catholic Church Philosopher, mathematician, jurist and astronomer He planned but never carried out an experiment to determine whether or not plants consume the soil He proposed they did not Revolutionary!! Jean Baptiste van Helmont (15791644) Flemish physician and chemist Identified carbon dioxide, carbon monoxide, nitrous oxide and methane He was a doctor. He married a wealthy noblewoman and her inheritance enabled him to retire early from medical practice and concentrate on his chemical experiments Over 5 years, he carried out experiment originally planned by Nicholas of Cusa and concludes the increase in mass of the plant came from water. He does, however, ignore a slight decrease in soil mass Robert Hooke Invented the light microscope Observed both plant and animal cells ‘Stoma’- from the Greek word for mouth First observed by Malphighi Stoma were so named by Heinrich Link because of their appearance Their function was unknown to him though Edme Mariotte (16201684) French physicist and priest In 1660 he discovered the eye’s blind spot! In 1676 he hypothesised that plants synthesise their food from air and water Stephen Hales (1677-1791) Physiologist, chemist and inventor He studied the roles of air and water and their importance to plant and animal life He wrote that plant leaves “very probably“ take in nourishment from the air and that light may also be involved Charles Bonnet Observed the emission of gas bubbles by a submerged illuminated leaf (clearly his pondweed was healthier than the pondweed we have in school!) Joseph Priestley and his experiments… 1733-1804 Theologian, philosopher, clergyman, scholar and teacher One of the scientists credited with discovering "dephlogisticated air“ – oxygen Finds out that air which has been made ‘noxious’ by the breathing of animals or burning of a candle can be restored by the presence of a green plant Carried out a very famous experiment using bell jars, candles, plants and mice… Antoine Lavoisier 1743-1794 Investigated and later named oxygen Recognises it is used up in both combustion and respiration His work discredits “phlogiston”, a hypothetical substance previously believed to be emitted during respiration or combustion One of the fathers of modern day chemistry Jan Ingenhousz 1730-1799 Physicist, chemist and plant physiologist Discovered photosynthesis (and Brownian motion!) Showed that light is essential for photosynthesis and that only the green parts of the plants release oxygen 1782 – Jean Senebier demonstrates that green plants take in carbon dioxide from the air and emit oxygen under the influence of sunlight 1791 – Comparetti observes green granules in plant tissues, later identified as chlorophyll Nicolas de Saussure 1767-1845 Chemist and plant physiologist Proved that the carbon assimilated from atmospheric carbon dioxide cannot fully account for the increase of dry weight in a plant The basic equation for photosynthesis was therefore established The Biochemistry begins… So scientists had now worked out that Carbon Dioxide was taken in and Oxygen was given out, and that the green pigment (named chlorophyll in 1818) played a part in this process, but what actually went on inside the leaf?... 1842 – Schleiden states that he believes the water molecule is split during photosynthesis 1844 – Hugo von Mohl makes detailed observations about the structure of chloroplasts 1845 – Julius Robert von Mayer proposes that the Sun is the source of energy used by living organisms and introduces the concept that photosynthesis converts light energy into chemical energy 1862 – Julius von Sachs demonstrates that starch formation in chloroplasts is light dependent The discoveries continue… 1864 – We have the balanced equation for photosynthesis after accurate quantitative measurements of carbon dioxide uptake and oxygen production are made… 6CO2 + 6H2O C6H12O6 + 6O2 1873 – Emil Godlewski proves that atmospheric CO2 is the source of carbon in photosynthesis by showing that starch formation in illuminated leaves depends on the presence of CO2 Not just any old light.. In 1883, Engelmann illuminated a filamentous alga with light that had been dispersed using a prism He discovered that aerobic bacteria in the water all congregated around the portions iluminated with red and blue wavelengths This was the first action spectrum! Plant Pigments and Chromatography Thin layer chromatogram (TLC) of an extract of thylakoid membranes from the leaf of annual meadow grass Poa annua. TLC plastic sheets are coated with a 60 F254 silica gel which measures 0. 2 millimetres thick. A drop of extract, corresponding to the column here, was laid at the bottom of the sheet. The sheet was then placed in a beaker of solvent (75% acetone & 25% petroleum ether). The picture shows the solubility of the extract in solvent. Six bands are seen; top (orange) is carotene; 2 (green) pheophytin; 3 (green) chlorophyll A; 4 (green) chlorophyll B; 5 (yellow) & 6 (mere trace) are carotenoids. The line across the top of image is the solvent line Solvent line Carotene Pheophytin Chlorophyll A Chlorophyll B Carotenoids Chlorophyll Chlorophyll + Light = Chlorophyll+ + Electron- Chlorophyll Found within chloroplasts Absorb and capture light Made up of a group of five pigments Chlorophyll a Chlorophyll b Carotenoids; xanthophyll and carotene Phaetophytin Chlorophyll a is the most abundant Proportions of other pigments accounts for varying shades of green found between species of plants Photosystem I and Photosystem II These are distinct chlorophyll complexes Each contains a different combination of chlorophyll pigments PSI absorbs light at 700nm and PSII at 680nm PSI particles are found on the intergranal lamellae PSII particles are found on the grana Photosynthetic Pigments Plants have a variety of different plant pigments. Each of these have a different absorption spectra enabling the plant to harvest a wide variety of different wavelengths of light. Chlorophyll a has two peaks of absorption in the blue and red end of the spectrum, it does not absorb strongly in the green wavelengths and as a result these wavelengths are reflected and the plant will appear green. The pigments are found in the grana of the chloroplast and arranged to maximise the absorption of light 1905 – Limiting Factors F.F. Blackman develops the concept of limiting factors He shows that photosynthesis consists of two stages… A rapid light dependent process and a slower temperature dependent process These become known as the ‘light’ and ‘dark’ reactions 1941 – Ruben and Kaman They set out to discover the path of carbon dioxide during photosynthesis but end up discovering something different… They experiment using heavy isotopes to discover whether the oxygen produced during photosynthesis comes from the splitting or water or carbon dioxide They discover water is split during the first, lightdependent stage of photosynthesis Daniel Arnon 1910-1994 Plant physiologist 1954 – he demonstrates light dependent ATP formation in chloroplasts 1955 – he demonstrates that isolated chloroplasts are capable of carrying out complete photosynthesis Chloroplasts Chloroplasts Thylakoids { Grana Intergranal lamellae Stroma Vacuole Cell wall Cytoplasm Grana False-colour transmission electron micrograph (TEM) of a stack of grana (black threads) in a plant chloroplast. The chloroplast is the unit within the leaf, which manufactures the food supply (starch) of the plant during photosynthesis. The granal stacks (flattened vesicles) contain the photosynthetic pigments (chlorophylls), which are active in the conversion of the sun's energy into chemical energy. The grana are connected at points called frets (the central conglomeration of black threads), which are embedded in the matrix of the chloroplast. The Light Dependent phase Is the first stage of photosynthesis. Where? The Thylakoid membrane (photosystems) Why? This is where chlorophyll and accessory pigments are. Photosystems. Purpose of the Photosystems is to trap light energy and convert it into Chemical energy in the form of ATP. Photosystem I was the first to be isolated but is actually the second stage. It is located mainly in the intergranal Lamella. Photosystem II was the second to be isolated but is the first stage of the reaction. It is located in the Granal lamella. Equation for photosynthesis. 6H2O + CO2 C6H12O6 +6 O2 Water is needed for photosynthesis but why? Photosystem II (granal) contains enzymes which split water into H+ ions (Protons), electrons and O2. This is known as photolysis. 2H2O + 4H+ + 4e- + O2 The ions which are produced are used for the Light Independent phase (Dark reaction) Some O2 is used for respiration PSI (P700) When light hits the chlorophyll molecule the light energy is transferred to the two electrons. These electrons become excited, and break their bonds. The electrons are captured by Electron acceptors and passed along a series of electron carriers. As electrons pass along this chain energy is released. This energy is used to pump protons across the thylakoid membrane into the Thylakoid space where they build up. As protons build up a gradient is created the protons flow down this gradient. This Process is known as Chemiosmosis. This process enables ADP and Pi to make ATP, which is used in the Light Independent Phase. (It could also be used by the guard cells to bring in K+ causing water to flow into the cell via osmosis and the stoma to open). The production of ATP via this method is known as Cyclic Photophosphorylation. Non Cyclic Photophosphorylation takes place in both the photosystems. It includes the production of ATP and NADP. Plant biologists are prize winners! 1956 – Melvin Calvin and his coworkers are awarded the Nobel Prize in 1961 after they use radioactively labelled CO2 to show the pathway of carbon assimilation during photosynthesis. The second stage of photosynthesis is also known as the Calvin Cycle! 1960 – Robert Woodward synthesises chlorophyll and is awarded the Nobel prize in 1965 1984 – Deisenhofer, Michel and Huber crystallise the photosynthetic reaction centre from a purple bacterium and use x-ray diffraction techniques to determine its detailed structure. They are awarded the Nobel Prize in 1988. Chloroplast 2. Oxidation and reduction OIL RIG = “Oxidation Is Loss (of electrons), Reduction Is Gain (of electrons) Any chemical which gives away electrons (e-) is said to be oxidised, and any chemical which accepts electrons is said to be reduced. 2. Oxidation and reduction In addition: Any chemical which gives away protons (H+) is oxidised, and any chemical which accepts protons is reduced H+ Chemical X reduction Chemical X e- Chemical X e- oxidation H+ Chemical X NAD NAD Function Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP) are two important cofactors found in cells. NADH is the reduced form of NAD+, and NAD+ is the oxidized form of NADH. It forms NADP with the addition of a phosphate group to the 2' position of the adenosyl nucleotide through an ester linkage. NAD is used extensively in glycolysis and the citric acid cycle of cellular respiration. The reducing potential stored in NADH can be converted to ATP through the electron transport chain or used for anabolic metabolism. ATP "energy" is necessary for an organism to live. Green plants obtain ATP through photosynthesis, while other organisms obtain it by cellular respiration. Photosynthesis 1. Light stage 2. Light Independent stage I. I. Takes Place in the Grana. II. Uses light energy. III. It makes ATP and reduced NADP. IV. Water is split in photolysis. V. Oxygen is released as a waste product. Takes Place in the Stroma. II. Uses ATP and reduced NADP from the light stage. III. It uses CO2 IV. It makes organic molecules such as sugars. Complicated diagram Less Complicated diagram The Light Stage High Reduce d NADP Energy Potential Light harvesting antennae ATP Photophosphorylation PSI Light harvesting antennae PSII Low electrons ADP electrons 1.Photosystem The light stage ofsome 3. IIupis now oxidised 4. Having given of its photosynthesis takes place in the and to replace the electron lost by energy the electron that passed grana of the chloroplast. Light the chlorophyll water is split in the down the electron transfer chain energy is byThis pigments process ofabsorbed photolysis. to photosystem I is excited again which are arranged into structures releases an electron a The by light energy givingand energy. called the light harvesting Hydrogen ion (H+). This electron electron and the hydrogen ion antennae. These funnel reduces chlorophyll inthe PSII. from the the photolysis of water energy towards a central core Oxygen istoreleased from the water combine reduce NADP. called photosystem as a waste product ofII.the process and some of will diffuse outlight of Therefore twothis products of the the plant. stage are ATP reduced of 2. Electrons in and the chlorophyll NADP. Oxygen is aexcited waste by the photosystem II are product. light energy and the chlorophyll oxidised. The excited electron is passed to a series of carriers and the energy is used to generate ATP, the process is called photophosphorylation. NADP H2O = OH- + H+ Photolysis 4OH = 2H2O + O2 Excretion Light Independent stage Energy from ATP, hydrogens and electrons from reduced NADP are used to reduce carbon dioxide to produce carbohydrate. This happens in the stroma. Complicated diagram 2 Light independent stage NADP Reduced NADP Carbon dioxide light independent reaction ATP ADP + Pi glucose (5C) Ribulose Biphosphate CO2 (1C) ATP 2X Glycerate-3-Phosphate NADPH (3C) (3C) ATP 2X Triose Phosphate Glucose (6C) C3 Cycle The Light Independent Stage 1. C3 cycle takes 3. The Regeneration: The place triosein the stroma chloroplast. The sugars of arethe converted in several cycle important steps has backthree to RUBP. This stages. process Carboxylation: The sugars pentosetosugar requires the triose be ribulose 1:5 bisphosphate phosphorylated by ATP. combines with carbon dioxide to Everyx2three times cycle sugar goes form of the threethe carbon around three carbons(GP). will be Glycerate phosphate This is addedwith andthe one surplus done help of thetriose enzyme sugar will be made. The surplus Rubisco. triose sugars are used to make other organic molecules. It will 2. Reduction: reduced take six turns The of the cycle toNADP is oxidised and Glyceratenew produce one completely phosphate molecule ofreduced glucoseand andconverted twelve to triose sugars (GALP). In foranother the disaccharide sucrose. this stepacids ATP also is converted to ADP Amino require the and phosphate, the ATP providing addition of nitrogen. activation energy for the process. CO ATP 2 ADP + Pi GP Reduce dNADP 3c RUBISCo NADP RUBP 3c 5c Regeneration ADP TP GALP 6c Glucose ATP Sucrose Light GP RUBP Dark The Light Independent Stage CO2 GP RUBP Glucose TP Glucose ATP Light Stage ATP Reduced NAD The Light Independent Stage CO2 GP RUBP Glucose TP Glucose ATP Light Stage ATP Reduced NAD What happens without light? Which stages can still continue when the light is turned off? Which molecules will build up when the light is turned off? Exam question Suggest why after the light was switched off the amount of GP... a) Increased immediately b) levelled out after a time Sketch the curve to show what happens to the amount of RuBP after the light has been switched off, Explain your answer. Sketch what would happen if CO2 was removed with the light left on. The light dependent reaction takes place on the... This is so that... The reaction needs ...., ...., .... and .... The reaction produces .... and ... .... is also produced. The ... and ...pass into the ... for the second stage in photosynthesis. 1. To understand the stages in aerobic respiration: glycolysis, link reaction, Kreb’s cycle and the electron transport chain 2. To link the stages 3. To explain the link between ATP production and energy levels Respiration Energy is released in respiration A series of oxidation reactions taking place inside living cells which releases energy to drive the metabolic activities that take place in cells Aerobic respiration – takes place in the presence of oxygen Anaerobic respiration – takes place in absence of oxygen Aerobic respiration –– to release energy 4 main stages CO2 glucose Krebs cycle Glycolysis FADH2 NADH pyruvate Link reaction Electron transport chain Hydrogen atoms Acetyl coenzyme A oxygen water Glycolysis -the splitting of glucose The phosphate comes from ATP 1. Glucose (6C) phosphorylated to Glucose phoshate (6C) 3. Glucose phosphate (6C) phosphorylated to fructose biphosphate (6C) 4. Fructose biphosphate (6C) is split into two molecules of glycerate 3 phosphate 5. Each Glycerate 3 – phosphate (3C) is converted to pyruvate (3C) 7. 2 x 2 ATP produced 6. H+ is removed and transferred to the hydrogen acceptor NAD (nicotinamide adenine dinucleotide) Glycolysis in detail Takes place in cytoplasm of cells Does not need oxygen – first stage of aerobic respiration and only stage of anaerobic respiration Although glycolysis yields energy it does need an input of energy to get the reaction started Glycolysis – overview Glycolysis produces from 1 molecule of glucose: 2 molecules of ATP in total (4 ATP are produced but 2 are used at the start) 2 molecules of NADH2 (reduced NAD) 2 molecules of pyruvate to enter the link reaction The link reaction in mitochondria in presence of oxygen Pyruvate (3C) 1. Pyruvate decarboxylated - CO2 removed NAD+ CO2 NADH + H+ 2. Pyruvate dehydrogenated – hydrogen removed Acetate (2C) Coenzyme A 3. Acetate (2C) combines with coenzyme A Acetyl coenzyme A Don’t forget this happens TWICE as 2 molecules of pyruvate are formed from each glucose molecule Krebs cycle in matrix of mitochondria