Metabolism och teknisk mikrobiologi Flashcards
Describe the citric acid cycle in detail. What are the substances/enzymes and end products?
Reaction pathway:
(Acetyl-CoA) -> Citrate -> Isocitrate -> Ketoglutarate -> Succinyl-CoA -> Succinate -> Fumarate -> Malate -> Oxaloacetate -> Citrate
Enzymes:
Citrate synthase -> Aconitase -> Isocitrate dehydrogenase -> Ketoglutarate dehydrogenase -> Succinyl-CoA synthetase -> Succinate dehydrogenase -> Fumarase -> Malate dehydrogenase -> Citrate synthase
End products:
Acetyl-CoA + Oxaloacetate + H2O -> Citrate + CoA-SH
Isocitrate + NAD -> Ketoglutarate + CO2 + NADH +H //
Ketoglutarate + NAD + CoA-SH -> Succinyl-CoA + CO2 + NADH +H //
Succinyl-CoA + GDP + P -> Succinate + CoA-SH + GTP //
Succinate + FAD -> Fumarate + FADH2 //
Fumarate + H2O -> Malate //
Malate + NAD -> Oxaloacetate + NADH + H //
In total: 3 NADH + 1 FADH2 + 2 CO2 - 2 H2O per Acetyl-CoA
Tip:
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Describe the Glyoxylate cycle in detail. What are the substances/enzymes and end products?
Reaction Pathway:
Citrate -> Isocitrate -> Glyoxylate + Succinate (path diverge) -> Glyoxylate + Acetyl-CoA -> Malate -> Oxaloacetate
Enzymes: Citrate -> Aconitase -> Isocitrate lyase -> Malate synthase -> Malate dehydrogenase
End products:
2 NADH from 2 Malate -> Oxaloacetate//
1 FAD + Succinate -> Fumarate + FADH2
In total: 2 NADH + 1 FADH2
Tip:
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Name substances that regulate TCA
Succinyl-CoA blocks steps 1 and 4.
NADH blocks 1, 3 and 4.
Anaplerotic
Anaplerosis is the act of replenishing the TCA cycle intermediates. Anaplerotic reactions are reactions that “fill up” intermediates in TCA cycle.
Describe each steps in Oxidative Phosphorylation and its end products.
Elelctron pair from NADH -> Complex 1 (4 H+ are pumped)
Electron pair reduces Ubiquinone -> Ubiquinol takes electron to Complex 3 (4 H+ are pumped)
Electrons pair travels to Complex 4 with 2 Cytochrome Cs. Complex 4 reduces oxygen with electron pair to form water (2 H+ are pumped) 2e + 4H+ 0.5 O2 -> H2O + 2H
Electron pair from FADH2 -> Complex 2 reduces Ubiquinone -> Ubiquinol takes electron to complex 3. Reaction continues as usual.
ATPase from ADP to ATP by protomotive force of proton gradient from intermembrane space to matrix. 1 turn creates 3 ATP. Usually 9 C-compartment in Eukaryotes mitochondria.
Explain how ATP, ADP and P are transported from mitochondria to cytolasm vice versa.
Outermembrane of mitochondria is freely permeable for ATP, ADP and P but the innermembrane need ATP/ADP carrier and Phosphate translocase. Switching between ADP and ATP costs H+. H2PO4 switches with OH and is electroneutral.
Which chemical substance can block/uncouple Oxidative Phosphorylation?
Oligomycin can block ATPase => no more ATP + slowed oxygen consumption
2,4-dinitrophenol uncouples and allows for H ions to cross membrane to get inside matrix. => no more ATP + increased oxygen consumption.
Explain how NADH from glycolysis can be transported to the electron transport chain in mitochondria.
NADH cannot cross the inner membrane. Bind electron pair to dihydroxyacetone phosphate (DHAP) to become glycerol-3-phosphate and reduce it at G3P dehydrogenase which creates FADH2. Or reduce oxaloacetate to become malate which is used to create one NADH. Oxaloacetate passes membrane by transamination to become Aspartate (out) and glutamine (in).
Name 6 inhibitors of the electron transport chain.
Complex I: Rotenone, Amytal.
Complex III: Antimycin A.
Complex IV: cyanide, azide, carbonmonoxide
Describe the structure of chloroplasts and its compartments.
Stroma = inside of chloroplasts
Granum = stacked thylakoids
Stroma lamella = between thylakoid granum (bridge)
Thylakoid lumen = cytpoplasm inside thylakoid
Pigments are located inside thylakoid membrane, ex chlorophyll a and b + accessory pigments like beta-carotene and lutein.
+ Antenna pigments to lower energy to the right wavelength.
Describe photosynthesis in detail. (Light reaction)
- 4 photons excite 4e in PS II (P680).
- 2 H20 + OEC -> O2 + 4H + 4e (OEC = oxygen evolving complex). The 4e replenishes PS II
- 4e from PS II go through an electron transportation to cytochrome b6f complex.
- 8H are pumped from Cytochrome b6f complex.
- 4 photons excite 4 electrons in PS I (P700)
- Electrons from Cytochrome b6f replenishes PS I
- Electrons from PS I are used to reduce 2NADP to 2NADPH
Tylakoids usually have ATPase with 12 C-subunits. Therefore 4H is neeeed for 1 ATP. 12H /8 photons = 1.5 H per photon. 3 ATP / 8 photons = 0.375 ATP per photon.
Describe the alternative cyclic of electron flow through PS I and when it is used.
Is used when NADPH is abundant and NADP is in short supply.
1 photon excites PS I (P700) -> e goes trough electron transport chain to cytochrome b6f (2 H is pumped) and is then used to replenish PS I back.
Describe the Calvin cycle in detail. (Dark reaction)
- (fixation) CO2 + Ru-1,5-bisphosphate -> 2x 3-phosphoglycerate // (RuBisCo enzyme, Co = carboxylase-oxygenase)
- (reduction) 2x 3PG + 2 ATP -> 2x 1,3-bisphosphoglycerate + 2 ADP//
2x 1,3-BPG + NADPH -> 2x glyceraldehyde-3-phosphate + NADP + P (1/6 of GA3P goes to gluconeogenesis!) - (acceptor regeneration) rearrange GA3P to Ribulose-5-phosphate. then Ru5P + ATP -> Ru-1,5-bisphospate + ADP
Overall:
3 CO2 + 6 ATP + 3PG -> 6 1,3-BPG + 6 ADP
6 NADPH + 6 1,3-BPG -> 6 GA3P + 6 NADP + 6P ( 1 GA3P goes to gluconeogenesis)
5 GA3P -> 3 Ru5P
3 Ru5P + 3 ATP -> 3 Ru-1,5-BP + 3 ADP
Describe why RuBisCo is ineffective.
Rubisco can react with oxygen to make Ru1,5BP to 3PG and 2PG, 2PG is not used in any reaction and needs to be recycled. It evolved when there was lots of CO2, the reaction only happens at low CO2.
Name an additional way for plants to fix CO2.
Plants can fix CO2 in mesophyll cells which can then re-release CO2 into calvin cycle.
What are TAG and where are they found?
Triacylglycerol, is the most abundant type of lipids and are found in adipose tissue in cells called adipocytes.
How are fatty acids broken off from TAG?
Hormones activate a G-protein that subsequently activates adenylate cyclase. Adenylate cyclase turns ATP to cAMP + PP. cAMP binds to subunit R on Protein Kinase A. Subunit C of Protein Kinase A activates different lipases that detaches a fatty acid from the glycerol backbone. Glycerol can then be used in glycolysis/gluconeogenesis. Glycerol is converted to glucose in liver. Free fatty acid binds to albumin in blood.
How are fatty acids broken down?
- “Activation” of free fatty acid to fatty acyl:
R-COO + ATP + CoA-SH -> R-CO-S-CoA + AMP + PP - Fatty-acyl binds to carnitine and releases CoA-SH (happens thanks to CPT I enzyme):
R-CO-S-CoA + Carnitine -> R-CO-Carnitine + CoA-SH - Fatty acyl carnitine enters matrix and releases carnitine and rebinds to CoA-SH thanks to CPT II enzyme.
- Thereafter 2 carbons are release after each beta oxidation cycle:
Fatty acyl-CoA + FAD + H2O + NAD + CoA-SH -> Fatty acyl-CoA (-2 C) + FADH2 +NADH + H + Acetyl-CoA
What happens in beta oxidation of fatty acid when there is an uneven amount of carbon atoms?
Propionyl-CoA is left ( 3 carbon atoms) and is turned into Succinyl-CoA which can be used in TCA cycle.
What happens in beta oxidation of fatty acid when there is a unsaturated fatty acid?
Depend on position of double bond.
If C2 = C3, no need for FAD to dehydrate.
If C3 = C4, move double bond to C2 = C3 by enoyl-CoA isomerase.
How are fatty acids synthesized?
Acyl-KS + malonyl-ACP + 2 NAPDH + 2H -> Acyl-KS (+2C) + 2 NAPD + CO2 + H2O + ACP
How are Malonyl-ACP and Acetyl-KS synthesized?
Acetyl-CoA + ACP -> Acetyl-ACP + CoA-SH //
Acetyl-ACP + KS -> Acetyl-KS + ACP //
Acetyl-CoA +HCO3 + ATP -> Malonyl-CoA + ADP + P + H//
Malonyl-CoA + ACP -> Malonyl-ACP + CoA-SH//
How can fatty acids be desaturated at specifik positions?
Fatty acyl-CoA + O2 + 3H + NADH -> Fatty acyl-CoA (desat.) + H2O + NAD
How are TAGs synthesized?
G3P + 2 Acyl-CoA -> Diacylglycerol-3-phosphate + 2 CoA-SH //
Diacylglycerol-3-phosphate + H2O -> diacylglycerol + phosphate //
diacylglycerol + Acyl-CoA -> triacylglycerol + CoA-SH //