Metabolism:
Chapter 6 Metabolism: Fueling Cell Growth
Fundamental Tasks for Growth
♣ Method of ______________ ______________ to invest in
♣ Biosynthetic processes
♣ Transporting nutrients / other molecules
♣ Continual ___________________ of new cell components
♣ Cell walls, membranes, ribosomes, nucleic acids, other
♣ Replacing worn or damaged components
♣ Accumulating enough components so that cell can divide
(growth = increase in cell number)
Principles of Metabolism
♣ ______________ is coupled to ______________
♣ Energy from breakdown of molecules is invested into buildup of new molecules
♣ ______________ reactions are coupled to ______________ reactions
Energy
♣ ______________ energy – stored energy
♣ Stored due to position (rock on a hill, water behind a dam)
♣ Stored due to chemical bonds
♣ ______________ energy – energy of motion
♣ Potential energy ______________ to kinetic
♣ But some is lost as ______________
♣ (random energy = entropy)
♣ Kinetic energy ______________ to potential
♣ Again, some is lost as ______________
♣ Lost energy must be replaced
Methods of Acquiring Energy
♣ Phototrophs
♣ Harvest energy in __________________
♣ Power synthesis of organic molecules
♣ e.g. glucose
♣ Chemoorganotrophs
♣ Harvest energy from __________________ molecules
♣ Depend on __________________ to synthesize high energy organic molecules
Free Energy
♣ __________________ energy released when bonds are broken
♣ Does not include heat
♣ __________________ reactions
♣ If more free energy in reactants
♣ __________________ reactions
♣ If more free energy in products
♣ Multiple ___________ allows slow release of free energy
♣ Less is lost as heat
♣ “Metabolic pathways”
Metabolic __________________
♣ Ordered sequence of steps resulting in an end-product
♣ Starts with reactants
♣ Continues with series of __________________
♣ May be in dissociated form (e.g. pyruvate)
♣ May be in undissociated form (e.g. pyruvic acid)
♣ Ends with __________________ __________________
♣ Pathway may be _________________, _________________, or _______________
Key Components of Metabolic Pathways
♣ Enzymes
♣ ATP
♣ Energy sources
♣ Redox reactions
♣ Electron carriers
♣ Precursor metabolites
Enzymes
♣ Enzymes ______________ on substrates
♣ Turn substrates into products
♣ Facilitate reactions – “__________________”
♣ Do not participate directly in reaction
♣ Without enzymes, some catabolic reactions would go very slowly
♣ Enzymes speed up reaction
♣ Enzymes lower the __________________ of __________________
♣ Even exergonic reactions
ATP Couples Energy Production with Energy Investment
♣ ________ donates energy to reactions
♣ Cycles to ___________
♣ ADP accepts energy
♣ Cycles to ATP
♣ ADP + Pi
♣ Cell constantly recycles ATP
Phosphorylation
♣ Ways to add Pi to ADP
♣ __________________ -level phosphorylation
♣ __________________ / proton motive force
♣ __________________ phosphorylation
Using redox reactions
♣ _______________phosphorylation
♣ Photoautotrophs only
♣ Sources of energy for phosphorylation
♣ All require __________________ reactions
♣ Transfer 1 or more electrons from one substance to another
♣ Prokaryotes very versatile
♣ e.g. Glucose
♣ e.g. Ammonia
Redox Reactions
♣ When substance is __________________
♣ Loses an electron
♣ When substance is __________________
♣ Gains an electron
♣ Where electrons go, protons __________________
♣ Lose e- & H+ = lose H atom = oxidation = dehydrogenation
♣ Gain e- & H+ = gain H atom = reduction = hydrogenation
♣ If e- removed to e- carrier
♣ H+ may go, too
♣ H+ may go into aqueous solution
♣ Often ignore the H+ when writing biological reactions, but it’s there!
e- Carriers that Diffuse Easily
♣ NAD+ reduces to __________________
♣ FAD+ reduces to __________________
♣ NADP+ reduces to NADPH
♣ (reduced = becomes less positive)
♣ Reduced form has __________________ __________________
♣ NADH & FADH2 reduce other carriers
♣ Drives proton motive force
♣ Proton motive force drives ATP production
♣ NADPH reduces molecules in __________________
Precursor Metabolites
♣ __________________ in catabolic reactions
♣ Link catabolic reactions to __________________ reactions
♣ Some are broken down further
♣ Some are siphoned off for anabolic rxns.
♣ e.g. pyruvate
♣ Oxidized further
♣ Convert to alanine
Use of Precursor Metabolites by E. coli
Central Metabolic Pathways
♣ 3 key pathways to gradually oxidize glucose
♣ __________________
♣ “Embden-Meyerhoff pathway”
♣ “Glycolytic pathway”
♣ __________________ __________________ pathway
♣ ________ cycle
♣ “Tricarboxylic acid cycle”
♣ “Krebs cycle”
♣ “Citric Acid cycle”
Central Metabolic Pathways
♣ Glycolysis
♣ Most __________________ to initiate sugar breakdown
♣ End products are ATP & NADH & pyruvate
♣ __________________ into Krebs TCA cycle
♣ Pentose Phosphate pathway
♣ Alternative pathway to make pyruvate
♣ Operates in conjunction with glycolysis
♣ __________________ feeds into TCA cycle
♣ Usually feeds into biosynthesis pathways
♣ [Entner-Douderoff pathway]
♣ Add’l alternative to glycolysis
♣ Some bacteria
♣ Less ATP
Fate of Pyruvate
♣ __________________ to TCA cycle from glycolysis, Entner-Douderoff, or Pentose Phosphate
♣ Pyruvate ⋄ Acetate + CO2
♣ 3C ⋄ 2C + C
♣ __________________ CoA enters __________________ cycle
♣ 2 pyruvates for each glucose that started
Respiration
♣ Uses reducing power of __________________ and __________________ produced in glycolysis and transition step
♣ Powers __________________ phosphorylation
♣ Electrons from NADH and FADH2 enter e- transport
♣ ________ pumped to other side of membrane
♣ Produces proton __________________
♣ Drives proton motive force
♣ Terminal e- acceptor _________ ______ O2 or other
Catabolic Processes of Chemoorganoheterotrophs
♣ __________________ respiration – ___________ ATP
♣ __________________ respiration – ___________ ATP
♣ __________________– _____________ ATP
Energy Yields
♣ Energy yields theoretical, as __________________ may be siphoned off to biosynthesis
♣ Calculated approximately with a theoretical __________________
♣ __________________ -level phosphorylation slow
♣ Little ATP produced
♣ __________________ phosphorylation fast
♣ Much ATP produced
Energy Yields in Glycolysis
♣ Glucose (6C) + 2 NAD+ + 2 ADP + 2 Pi
♣ ⋄ 2 pyruvate (3C) + 2 NADH + 2 H+ 2 ATP (net gain)
♣ Also produces precursor molecules (used by E. coli)
Yields from Glycolysis
♣ Energy expended --- ______ ATP per glucose
♣ Energy harvested --- ______ ATP
♣ Net gain ------------- ______ ATP
♣ Reducing power ----- ______ NADH
♣ __________________ metabolites (reduce energy yield) --- ______ (based on E. coli) plus pyruvate
♣ ATP produced by ------- __________________ level __________________
Yields from Pentose Phosphate Pathway
♣ Energy yield ---- __________________ amts ATP
♣ Reducing power ---- __________________ amts NADH
♣ Precursor metabolites --- ______ plus pyruvate
♣ Most of the products of the Pentose Phosphate Pathway are used as precursors in biosynthesis
♣ ATP produced by ------- __________________ level __________________
Yields of the Transition Step and TCA Cycle
♣ Transition step (yield per glucose)
♣ Reducing power --- ______ NADH
♣ Precursor metabolites --- ______ Acetyl CoA
♣ TCA Cycle (yield per glucose)
♣ Energy yield --- ______ ATP
♣ GTP in step 5 converts to ATP
♣ Reducing power --- ______ NADH + ______ FADH2
♣ Precursor metabolites --- ______ (based on E. coli)
♣ ATP produced by __________________ level __________________
Electron Carriers in the Plasma Membrane
♣ Electron carriers embedded in the __________________ __________________
♣ Where in eukaryotes?
♣ Electron transport goes from ______________ energy to _________ energy carriers
♣ Some carry H+, too
♣ Some push H+ to other side of membrane
♣ FADH2 __________________ chain __________________ than NADH
♣ Less __________________ results from FADH2
♣ Terminal e- acceptor ________ ______ ½ O2
♣ If anaerobic, then ________ ½ O2
Oxidative Phosphorylation
♣ Chemiosmotic __________________ (proton motive force) drives ATP synthesis
♣ H+ flow back across membrane through ion __________________ in ________ __________________
♣ Called “__________________” even if O2 not terminal e- acceptor
♣ Still oxidizing!
♣ Very __________________ amounts of ATP made
♣ Variation in _______ __________________ allows differential testing
♣ Oxidase test detects cytochrome c
Theoretical Energy Yields from Oxidative Phosphorylation
♣ All theoretical
♣ Not precise conversion
♣ Siphoning off of precursor metabolites reduces yield from theoretical max
♣ Based on reducing power from other steps
♣ Assume ______ ATP / NADH
♣ Assume ______ ATP / FADH2
♣ Reducing power from:
♣ Glycolysis
♣ ______ NADH ⋄ ______ ATP
♣ Transition step
♣ ______ NADH ⋄ ______ ATP
♣ TCA cycle
♣ ______ NADH ⋄ _______ ATP
♣ ______ FADH2 ⋄ ______ ATP
♣ ________ ATP total
♣ Maximum for oxidative phosphorylation
Maximum Theoretical Energy Yields for Aerobic Respiration = ________ ATP
♣ Substrate level
phosphorylation – ______ ATP
♣ Glycolysis ⋄ ______ ATP
♣ TCA cycle ⋄ ______ ATP
♣ Oxidative
phosphorylation – ________ ATP
♣ Reducing power from glycolysis ⋄ ______ ATP
♣ Reducing power from transition step ⋄ ______ ATP
♣ Reducing power in TCA cycle ⋄ ________ ATP
Anaerobic Pathways
♣ Two types
♣ __________________ __________________
♣ __________________
♣ Both use glycolysis & pentose phosphate pathway
♣ Differ in __________________ e- __________________
♣ Differ in what happens to __________________
♣ Anaerobic respiration
♣ Terminal e- acceptor not O2 and not organic
♣ __________________ reducers (NO3-) or _________________ reducers (SO42-)
♣ Pyruvate oxidized to _____________
♣ Fermentation
♣ __________________ ⋄ terminal e- acceptor
Anaerobic Respiration
♣ __________________ (NO3-) reduced to nitrite (NO2-)
♣ Or to nitrous oxide (N2O)
♣ Or to nitrogen gas (N2)
♣ __________________ (SO42-) reduced to hydrogen sulfide (H2S)
♣ __________________ efficient than aerobic respiration
Fermentation
♣ Fermentation pathways used by
♣ __________________ __________________ (e.g. E. coli)
♣ __________________ species
♣ Lactic acid bacteria ferment only – never respire
♣ Obligate fermenters that are not harmed by O2
♣ __________________ __________________ that ferment
♣ Harmed by O2
♣ ATP comes only from __________________
♣ Other steps recycle NADH from NAD+
♣ __________________ or derivative used as e- __________________
End Products of Fermentation
♣ __________________ Acid
♣ Pyruvate is terminal e- acceptor
♣ e.g. Lactic acid bacteria
♣ Streptococcus & Lactobacillus spp.
♣ Yogurt, pickles, cheeses, cured sausages
♣ Acid ⋄ tooth decay
♣ Food spoilage
♣ __________________
♣ CO2 removed ⋄ EtOH + CO2
♣ e.g. Saccharomyces spp. (yeast)
♣ Wine, beer, bread
♣ __________________ Acid
♣ e.g. Clostridium spp.
♣ __________________ Acid
♣ CO2 removed ⋄ modification ⋄ propionate
– Propionibacterium spp
♣ Swiss cheese
♣ 2, 3 – __________________
♣ Differentiates members of Enterobacteriaceae
♣ e.g. Klebsiella spp., Enterobacter spp.
♣ Voges-Proskauer test detects acetoin
♣ Mixed __________________
♣ Various pathways branched
♣ Methyl red test differentiates E. coli from other Enterobacteriaceae
Catabolism of Other Compounds
♣ Compounds other than glucose
♣ Broken down by hydrolytic enzymes
♣ May then enter cycles at various points
♣ __________________
♣ Lipases break down to precursor metabolites
♣ Enter various pathways
♣ __________________
♣ Proteases break peptide bonds
♣ Deaminases remove amino group
♣ Precursor metabolites enter various pathways
♣ __________________
♣ Amylase breaks down amylose
♣ Cellulase breaks down cellulose
♣ e.g. fungi
♣ e.g. bacteria in rumen of many herbivores & termites
♣ β-galactosidase breaks down lactose ⋄ glucose & fructose
♣ Glucose enters glycolysis directly
♣ Other monosaccharides modified first
Categories of Organisms by Energy and Carbon Sources
Chemolithotrophs
♣ Use __________________ source of energy
♣ ________ – hydrogen sulfide
♣ ________ – ammonia
♣ Products of anaerobic respiration __________________
♣ Fix __________________ into organic molecules from CO2
♣ = Chemolithoautotrophs
Photoautotrophs
♣ Use photosynthesis to
♣ Harvest energy from __________________
♣ Fix __________ into organic molecules
♣ 6 CO2 + 12 H2X [pic] C6H12O6 + 12 X + 6 H2O
♣ ______ depends on source of electrons
♣ Two stages
♣ Light __________________– __________________
♣ Light __________________ (“dark”) – carbon __________________
Photophosphorylation
♣ Light energy harvested in __________________ complexes
♣ Many __________________ absorb light
♣ Pass __________________ to reaction-center chlorophyll
♣ Excited electrons pass along electron transport chain in membrane e- carriers
♣ __________________
♣ Generates proton motive force and _________
♣ _______________________
♣ Generates __________________ power
♣ NADPH generates reducing power for e- transport
Electron Sources in Photosynthesis
♣ Source of __________________ determines if __________ is produced
♣ __________________ photosynthesis
♣ Plants, algae, cyanobacteria
♣ __________________ is source of electrons
♣ O2 is generated
♣ __________________ photosynthesis
♣ Green and purple bacteria
♣ ___________, ________ gas is source of electrons (not water)
♣ _____ O2 is generated
Carbon Fixation
♣ Conversion of _________ to organic form
♣ __________________– requires input of much energy (__________)
♣ Several cycles
♣ Calvin cycle most common
♣ Uses ribulose bisphosphate carboxylase
♣ “__________________”
♣ __________________ abundant / important enzyme on earth
♣ 3 turns of the cycle
♣ ⋄ one glyceraldehyde–3–phosphate (___________, a 3-carbon sugar)
♣ 6 turns of the cycle
♣ ⋄ one molecule of 6-__________________ sugar (e.g. fructose 6-phosphate)
♣ Consume 18 ATP & 12 NADPH + H+
Calvin Cycle
__________________ Pathways
Using __________________ Molecules
♣ Variation in ability to utilize substrates
♣ Variation in ability to synthesize intermediates
♣ Basis of selective media
♣ Basis of differential media
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