Kreb’s Cycle

(aka, tricarboxylic acid

(TCA)cycle, citric acid cycle)
“The wheel is turnin’ and the sugar’s a
burnin’”
 Overall goal
• Makes ATP
• Makes NADH
• Makes FADH2
• Requires some carbohydrate to run
 Geography
• Glycolysis in the cytosol
• Krebs in mitochondrial matrix
• Mitochondrion
– Outer membrane very permeable
• Space between membranes called intermembrane space
(clever huh!)
– Inner membrane (cristae)
• Permeable to pyruvate,
• Impermeable to fatty acids, NAD, etc
– Matrix is inside inner membrane
 Conversion of pyruvate to Acetyl
CoA
NAD+ NADH HSCoA CO2 O
O
O SCoA
H3C H3C
O
pyruvate dehydrogenase complex acetyl CoA

pyruvate

• 2 per glucose (all of Kreb’s)

• Oxidative decarboxylation
• Makes NADH
• -33.4kJ
 Fates of Acetyl CoA
O
Kreb's
TAG's H3C SCoA
acetyl CoA CO2, ATP, NADH...energy

no CHO present

ketone bodies

• In the presence of CHO an using energy

– Metabolized to CO2, NADH, FADH2,GTP and, ultimately, ATP
• If energy not being used (Lots of ATP present)
– Made into fat
• If energy being used, but no CHO present
– Starvation
– Forms ketone bodies (see fat metabolism slides)
– Danger!
 Kreb’s Cycle
acetyl CoA
H3C SCoA
C
O O
O
CoASH C
O O O H2O
O + CH2 O
C
C NAD
NADH
HO C C O
C O citrate synthase
HC OH CH2
CH2
H2O CH2 malate C citrate
dehydrogenase
C O O
O O C O
O
fumarase O
C O oxaloacetate
C H malate
aconitase
H C fumarate

O
C
O Kreb's Cycle O O
FADH2 C
succinate
dehydrogenase HO CH O
HC C O
FAD alpha ketoglutarate CH2
O SCoA O C isocitrate
O NADH O O
C GTP NAD C
CoASH C NADH O O
CH2 CH2 C O CO2 NAD
CH2 GDP CH2
CH2 CoASH
C C CO2 CH2
succinyl CoA isocitrate dehydrogenase
O O synthetase O O alpha ketoglutarate C
succinate succinyl CoA
dehydrogenase
O
O
 Net From Kreb’s
• Oxidative process
– 3 NADH
– FADH2
– GTP
• X 2 per glucose
– 6 NADH
– 2 FADH2
– 2 GTP
• All ultimately turned into ATP (oxidative
phosphorylation…later)
 Citrate Synthase Reaction (First)
O O O
C O
C
C O H2O CoASH
CH2 CH2 O
H3C SCoA
C C HO C C O
O + O
O
citrate synthase CH2
C
O O
acetyl CoA oxaloacetate
citrate

• Claisen condensation
• -32.2kJ
 Aconitase Reaction
O O O O
C C
CH2 O
HO CH O
HO C C O HC C O
CH2 CH2
aconitase
C C
O O
O O

citrate isocitrate

• Forms isocitrate
• Goes through alkene intermediate (cis-aconitate)
– elimination then addition
• 13.3kJ
 Isocitrate Dehydrogenase
O O
O O C
C C O
HO CH O
NAD NADH CO2 CH2
HC C O CH2
CH2
C C
O O isocitrate dehydrogenase O O

isocitrate alpha ketoglutarate

• All dehydrogenase reactions make NADH or FADH2

• Oxidative decarboxylation
• -20.9kJ
• Energy from increased entropy in gas formation
 α-ketoglutarate dehydrogenase
O O
C SCoA O
C O
C
CH2 CO2 NADH
CoASH NAD CH2
CH2
CH2
C C
O O O O
alpha ketoglutarate
dehydrogenase
alpha ketoglutarate succinyl CoA

• Same as pyruvate dehydrogenase reaction

• Formation of thioester
– endergonic
– driven by loss of CO2
• increases entropy
• exergonic
• -33.5kJ
 Succinyl CoA synthetase
SCoA O O
O
C C
CH2 GTP CoASH CH2
GDP
CH2 CH2
C C
O O succinyl CoA O O
succinyl CoA synthetase
succinate

• Hydrolysis of thioester
– Releases CoASH
– Exergonic
• Coupled to synthesis of GTP
– Endergonic
– GTP very similar to ATP and interconverted later
• -2.9kJ
 Succinate dehydrogenase
O O O
O
C FADH2 C
CH2 FAD C H
CH2 H C
C succinyl CoA
C
O O
O O dehydrogenase

succinate fumarate

• Dehydrogenation
• Uses FAD
– NAD used to oxidize oxygen-containing groups
• Aldehydes
• alcohols
– FAD used to oxidize C-C bonds
– 0kJ
 Fumarase
O O O O
C H2O C
C H HC OH
H C CH2
C fumarase
C
O O O
O
fumarate
malate

• Addition of water to a double bond

• -3.8kJ
 Malate Dehydrogenase
O O
C O O
HC OH C
NAD NADH C O
CH2 CH2
C C
O O malate O
dehydrogenase O
malate
oxaloacetate

• Oxidation of secondary alcohol to ketone

• Makes NADH
• Regenerates oxaloacetate for another round
• 29.7 kJ
 Net From Kreb’s
• Oxidative process
– 3 NADH
– FADH2
– GTP
• X 2 per glucose
– 6 NADH
– 2 FADH2
– 2 GTP
• All ultimately turned into ATP (oxidative
phosphorylation…later)
 Total Energy per glucose
• Cytosol
– Glycolysis
• 2 NADH
• 2 ATP
• Mitochondrion
– Pyruvate dehydrogenase
• 2 NADH
• Krebs
– 6 NADH
– 2 FADH2
– 2 GTP
 Total Energy/glucose
• In mitochondrion:
– Each NADH makes 2.5 ATP
– Each FADH2 makes 1.5 ATP
– GTP makes ATP
• So…
– From in mitochondrion
• 8 NADH X 2.5 ATP/NADH = 20 ATP
• 2 FADH2 X 1.5 ATP/FADH2= 3 ATP
• 2 GTP X 1 ATP / GTP = 2 ATP
• TOTAL in mitochondrion 25 ATP
 Total Energy/ glucose
• Cytosol
– 2 ATP
– 2 NADH
• NADH can’t get into mitochondrion
• In eukaryotes two pathways,
– transferred to FADH2
» get 1.5 ATP/ FADH2
– Or transferred to NADH
» Get 2.5 ATP/ NADH
– (Not a problem in prokaryotes (why?))
– 2 NADH X 1.5 ATP = 3 ATP
– Or 2 NADH X 2.5 ATP = 5 ATP
» + =2 ATP
» Total 3+ 2 or 5 + 2 so either 5 or 7
 ATP/glucose
• Eukaryotes
– Mitochondrial: 25 ATP
– Cytosolic: 5 or 7 ATP
– Total 30 or 32 ATP/glucose
– 30 ATP X 7.3kcal X 4.18 kJ = 915 kJ
ATP kcal

If 32 ATP = 976 kJ
• Prokaryotes
– 32 ATP X 7.3kcal X 4.18 kJ = 976 kJ
ATP kcal