seminar iii - uni-due.de · chemiosmotic model • in this simple representation of the...
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Seminar III
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Seminar III
• ATP-Generation via substrate level
phosphorylation, electron transport
phosphorylation
• CO2 Fixation
• (Eukaryotic/prokaryotic cell)
• Bacterial cell wall
• Antibiotic resistance
2
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Basic Mechanisms of Energy Conservation
Fig. 5.13 Brock Biology of Microorganisms (11th edition) (Madigan et al.)
Elektron-transport phosphorylation
(Oxidative Phosphorylation)
Substrate-level phosphorylation
Formation of energy-rich intermediates
produces ATP
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Energy-Rich Compounds
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Chemiosmotic Model
• In this simple representation of the chemiosmotic theory applied to mitochondria, electrons from
NADH and other oxidizable substrates pass through a chain of carriers arranged asymmetrically
in the inner membrane.
• Electron flow is accompanied by proton transfer across the membrane, producing both a chemical
gradient (ΔpH ) and an electrical gradient (Δψ).
• The inner mitochondrial membrane is impermeable to protons; protons can reenter the matrix only
through proton-specific channels (Fo). The proton-motive force (PMF) that drives protons back
into the matrix provides the energy for ATP synthesis, catalyzed by the F1 complex associated
with Fo.
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PMF Energized Membrane
Proton-motive force (PMF)
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ATP-Synthase/ATPase
Fig. 14.15 Essential Cell Biology (2nd edition, Alberts, Bray et al.)
-reversibel
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Energetics of Carbohydrate Metabolism
(Aerobic Respiration)
Glucose
2 Pyruvate
2 Acetyl-CoA
2 CO2
4 CO2
GlycolysisATP2
ATP2Citric Acid
Cycle
2 NADH2
6 NADH2
2 NADH2
2 FADH2
Pyruvate
dehydrogenase
Aero
bic
respiration
chain
ATP18
ATP6
ATP6
ATP4
ATP38ATP34ATP4 + =
O2 as final
electron
acceptor
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Energetics Balance Aerobic Respiration
ATP-Synthese (∆G0‘=-31,8 KJ/mol)
Einschl. (1 NADH)
Pyruvate-Dehydrogenase
Komplex
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Aerobic Respiration „Eucaryotes“
Abb. 9.6 Die Zellatmung im Überblick.Biologie (Campbell)
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Regulation
Fig. 9.20 Die Kontrolle der Zellatmung.Biology (6th edition, Campbell & Reece)
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Conversion of different Nutrients
Fig. 9.19 Catabolism of different nutrients.Biology (6th edition, Campbell & Reece)
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„Platform Metabolism“
Anabolism
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Fermentation
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Atmung/Fermentation
Fig. 5.14 Microbiology: An Introduction (Tortora, Funke, Case)
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Lacitc Acid Fermentation
Lactococcus lactis
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Anaerobic Respiration
• Alternative electron acceptors in absence of oxygen
• Energy source:
mostly organic compounds (chemoorganotrophic org.); but also inorganic compounds (chemolithotrophic org.)
• Electron acceptors:
Inorganic compounds, NO3-, SO4
2-, Fe3+, NO2-, S0, CO2
• Electron transport chain:
analogue to the aerobic chain (Cytochrome, Quinone, Fe-S Proteine)
• Facultative aerobes/anaerobes with aerobic and anaerobic respiration;
• Obligate anaerobes only anaerobic respiration
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Nitrate Reduction (E. coli)
• Enterobacteriaceae (e.g. E. coli)
• Facultative anaerobic Bacteria (anaerobic fermentation)
• Only reduction of nitrate to nitrite (nitrate reductase A)
aerob anaerob (NO3-)
Fig. 17.37 Vergleich aerobe und Nitrat Atmung. Brock Biology of Microorganisms (10th edition) (Madigan et al.)
½ O2/H2O +0,82 V NO3-/NO2
- +0,43 V
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Knallgas-Bacteria
• Biologic knallgas-reaktion
„Oxidation of hydrogen“
H2 + ½ O2 → H2O ΔG0‘=-237 kJ
„Hydrogenase“
• Different Bacteria:
G-: Pseudomonas, Alcaligenes,
Paracoccus,
G+: Nocardia, Mycobacterium,
Bacillus
• Hydrogenase (membrane-bound)
„Electron transport“; some
organisms in addition soluble
hydrorgenase „direct reduction of
NAD+“
• Chemolithoautotrophe „CO2
fixation via calvin cycle“
• Chemoorganotrophic growth
(Calvin cycle and hydrogenase
repressed)
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Photosynthesis
Light- and Dark- Reaction
Fig. 10.4 Biology (6th edition, Campbell & Reece)
Calvin cycle
Melvin Calvin (1940)
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Non-cyclic Photophosphorylation
Fig. 10.12 Biology (6th edition, Campbell & Reece)
P700 absorbs dark red light (700nm)
P 680 absorbs red light (680 nm)
Plastoquinone (Pq), Cytochrome b6-f-complex (proton pump), Plastocyanin
(Pc, Cu2+-Protein)
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The Light Reaction and Chemiosmosis
Fig. 10.16 Biology (6th edition, Campbell & Reece)
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Comparison of Chemioosmosis in
Mitochondrion and Chloroplast
Fig. 10.8 Biology (6th edition, Campbell & Reece)
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CO2 Fixation
CO2 fixation pathway ATP
requirement/
pyruvate
Relation to O2 Advantages
Calvin cyle 7 ATP aerobes Products = sugars,
seperated from other
metabolic pathways
Reductive citric acid cycle 2(-3) ATP anaerobes,
microaerobes
Suited for microaerobic
conditions, revers:
Oxidation of acetyl-CoA
Reductive acetyl-CoA
pathway
1 ATP strict
anaerobes
Suited for the assimilation
of C1 units
3-hydroxypropionate
pathway
7 ATP aerobe Suited for mixotrophic
assimilation of
fermentation products
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The Calvin Cycle „Dark-Reaction“
Fig. 10.17 Biology (6th edition, Campbell & Reece)
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Key Reactions of the Calvin Cycle (Reductive Pentose Phosphate Cycle)
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Balance of the Calvin Cycle
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Reductive pentose phosphate cycle(Calvin-Benson cycle)
2 Pyruvat
4 ATP2 NAD(P)H
RubisCO = Ribulose-bisphosphate carboxylase/oxygenase
7ATP +5 NAD(P)H/pyruvateΣ
• Plants, algae, cyanobacteria, most aerobic and facultativ aerobic Bacteria• Triosephosphates, 3-phosphoglycerate, sugar phosphates as intermediates
From light dependentreactions ofphotosynthesis(ETP reactions in chemosynthesis)
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Photosynthesis
Light- and Dark- Reaction
Fig. 10.4 Biology (6th edition, Campbell & Reece)
Calvin cycle
Melvin Calvin (1940)
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Pyruvate Dehydrogenase and
Oxidative Citric Acid Cycle
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Oxidative Citric Acid Cycle
Fig. 9.11 The citric acid cycle.Biology (6th edition, Campbell & Reece)
Aconitase
Citrate synthase
Isocitrate
dehydrogenase
α-ketoglutarate
dehydrogenase complex
Succinate
dehydrogenase
Succinyl-CoA
synthetase
Malate
dehydrogenase
Fumarase
Condensation
Dehydration/Hydration
Oxidative
decarboxylation
Oxidative
decarboxylation
Substrate-level
phosphorylation
Dehydrogenation
Hydration
Dehydrogenation
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Reductive Citric Acid Cycle(Arnon-Buchanan cycle)
2 (-3) ATP + 3 NAD(P)H + 2 ferredoxin/pyruvateΣ
• Anaerobic Green sulfur bacteria (Chlorobiales) and other Proteobacteria, Aquificales(microaerophilic)
• Acetyl-CoA, pyruvate, oxaloacetate, succinyl-CoA, 2-oxoglutarate, (PEP)• Advantages under anaerobic, microaerophilic conditions
2-Oxoglutarate synthase
ATP-citratelyase
Pyruvatesynthase
Isocitratedehydrogenase
Fumaratereductase
ATP and ferredoxin fromlight dependentreactions ofphotosynthesis(ETP reactions in chemosynthesis)
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Reductive Acetyl-CoA PathwayWood-Ljungdal pathway
~ 1 ATP + 2 NAD(P)H (F420H2) + 3 ferredoxin/pyruvateΣ
• Strict anaerobes: Methanogenic and sulfate reducing Euryarchaea, acetogens, some proteobacteria a.o.
• Acetyl-CoA, pyruvate• Well suited for the assimilation of C1 units
Acetyl-CoA synthase/CO dehydrogenase
Pyruvat
CO2 + Fdred
Fdox
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3-Hydroxypropionate Bi-Cycle
2x
~ 7 ATP + 6 NAD(P)H (- 1QH2) /pyruvateΣ
Acetyl-CoAcarboxylase
Propionyl-CoAcarboxylase
• Anaerobic Chloroflexaceae• Acetyl-CoA, pyruvate, succinyl-CoA• Suited for mixotrophic assimilation of fermentation products
Malyl-CoAlyase
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Hydroxypropionate Bi-Cycle
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Dicarboxylate/4-Hydroxybutyrate cycle3-Hydroxypropionate/4-Hydroxybutyrate cycle
Dicarboxylat/4-Hydroxybutyratecycle
3-Hydroxypropionate/4-Hydroxybutyrate
cycle
AnaerobicCrenarchaeota(Thermoproteales, Desulfurococcales
Aerobic Crenarchaeota (e.g.
Sulfolobus) andThaumarchaeota
(Thermoproteales, Desulfurococcales
~ 5 ATP + 2 Fdred + NAD(P)H + 4 [H] /pyruvateΣ ~ 6 ATP + 3 NAD(P)H + 4 [H] /
pyruvateΣ
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Das griechische Alphabet
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Seminar III
• ATP-Generation via substrate level
phosphorylation, electron transport
phosphorylation
• (Eukaryotic/prokaryotic cell)
• Bacterial cell wall
• Antibiotic resistance
40
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Prokaryotes & Eukaryotes
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Murein: Cell wall (Bacteria)
• Gram positive cell wall
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Murein: Cell wall (Bacteria)
• Gram negative cell wall
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Glycoconjugate
• Glycolipids
– Lipopolysaccharide (Gram negative Bacteria,
outer membrane)
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N-acetylglucosamine
• N-acetylglucosamine, a sugar derivative,
basic building block for chitin and murein.
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Hexose/Zuckerderivate
• Hydroxylgruppen in der
Ausgangsverbindung sind
durch andere Substituenten
ersetzt . Aminogruppe (z.B.
Glucosamin)
– Aminogruppe kondensiert mit
Essigsäure (N-
Acetylglucosamin)
– Milchsäure verbunden mit C-
4 Atom N-Acetyl-
muraminsäure
– Substitution einer
Hydroxylgruppe durch
Hydrogen (z.B. Fucose)
– Oxidation einer Aldehyd-
gruppe zur Aldonsäure (z.B.
Gluconsäure)
– C-6 Oxidation Uronsäure
(z.B. Glucuronsäure)
– Sialinsäure C-9 Zucker
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Murein: Cell wall (Bacteria)
• Structure of the polysaccharide
in bacterial cell wall
peptidoglycan.
The glycan is a polymer of
alternating GlcNAc and N-
acetylmuramic acid (MurNAc,
Lactic acid linked to C-4 atom)
residues.
Alternating peptide chains of D-
und L-amino acids
Linkage of the L-alanine amino
group (amide linkage) with the
lactylcarboxylgroup of a MurNac
residue
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Murein: Cell wall (Bacteria)
(a) Diaminopimelinsäure
(b) Lysin
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Murein: Cell wall (Bacteria)
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Figure 27.12
Mode of Action of Some Major
Antimicrobial Agents
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Figure 27.13
Antimicrobial Spectrum of Activity
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Antimicrobial Drug Resistance
• Antimicrobial drug resistance
– The acquired ability of a microbe to resist the effects of
a chemotherapeutic agent to which it is normally
sensitive
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Mechanisms of Drug Resistance
• Prevent entrance of drug
• Drug efflux (pump drug out of cell)
• Inactivation of drug
– chemical modification of drug by pathogen
• Modification of target enzyme or organelle
• Use of alternative pathways or increased production of target metabolite
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Antimicrobial Drug Resistance
• At least six reasons that microbes are naturally
resistant to certain antibiotics
– Organism lacks structure the antibiotic inhibits
– Organism is impermeable to antibiotic
– Organism can inactivate the antibiotic
– Organism may modify the target of the antibiotic
– Organism may develop a resistant biochemical
pathway
– Organism may be able to pump out the antibiotic
(efflux)
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Figure 27.27
Sites at Which Antibiotics are Attacked by
Enzymes
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Mechanisms of Bacterial Resistance to Antibiotics
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Antimicrobial Drug Resistance
• Most drug-resistant bacteria isolated from patients
contain drug-resistance genes located on R plasmids
• The use of antibiotics in medicine, veterinary, and
agriculture select for the spread of R plasmids
– Many examples of overuse of antibiotics
– Used far more often than necessary (i.e., antibiotics used
in agriculture as supplements to animal feed)
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Figure 27.28 Figure 27.28
Patterns of Drug Resistance in Pathogens