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Metabolic Engineering UCSD
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Metabolic BiochemistryBIBC102
Lecture 10
October 24, 2014
Metabolic flux and its control
1 Glucose (C6H12O6)
2 GAP
2 PYR
2 LACTIC ACID ( 2 x C3H6O3)
2 ADP + 2 Pi
2 ATP
Biological Work
Feedback Inhibition
LNC 15.16 (3rd ed.)
“bottleneck”)
We are not dealing with isolated reactions
at equilibrium, but with a flux of carbons
through a pathway
this flux must be regulated and adjusted to
physiological needs
near equilibrium
LNC 3rd ed.
LNC 15.18a
active site
active site
regulatory site
regulatory site
Phosphofructokinase 1
(only two of four subunits shown)
Fru 6-P + ATP Fru 1,6-bisphosphate + ADP
LNC 15.18b
ATP is both a substrate
anda negative modulator
LATERLecture 20
AMP stimulates the enzyme !!!!ATP inhibits the enzyme !!!!
High AMP
Low AMP
AMP concentrations
fluctuate more widely, and therefore the sensitivity
of the enzyme can be increased
if it responds to changing
AMP concentrations
The role of adenylate kinase:
2 ADP ATP + AMP Keq = 0.44
[ATP] [AMP]Keq = 0.44 =
[ADP]2
when [ATP] is high, [AMP] is low
when [ATP] is low, [AMP] is high
LNC 4th ed.
Typical physiological concentrations
A relatively small decrease in ATPcauses
a relatively large increase in AMP
A researcher (Catalina Island Marine Institute) scuba diving in the Toyon Bay found this dead fish that normally
lives 3000 feet below the surface.
We do not normally eat GLUCOSE,
but we eat carbohydrates/junk food
in various forms
Other Common Sugars
Monosaccharides: fructosegalactosemannose
Disaccharides: sucrose Glu-Frulactose Gal-Glumaltose Glu-Glutrehalose Glu-Glu
(Section 14.2) Feeder Pathways for Glycolysis)
Disaccharides
Figure 7-12
Galactose Glucose
Figure 7-12
GlucoseFructose
Figure 7-12
Glucose Glucose
Formation of an ETHER
R1 - CH2 - O - H + H - O - CH2 - R2
R1 - CH2 - O - CH2 - R2
H2O
Lactose + H2O D-galactose + D-glucose
Sucrose + H2O D-fructose + D-glucose
Trehalose + H2O 2 D-glucose
Lactase
Sucrase
Trehalase
Enzymes on surface of intestinal epithelial cells
Lactose IntoleranceOMIM
Online Mendelian Inheritance in Man
Major symptoms: diarrhea, dehydration, bloat
Remedy: Lactaid (lactase) taken orally
http://www.ncbi.nlm.nih.gov/omim?cmd=search
In humans, the activities of lactase and most of the other digestive hydrolases are maximal at birth. The majority of the world's human population experiences a decline in production of the digestive enzyme lactase-phlorizin hydrolase during maturation, with the age of onset ranging from the toddler years to young adulthood. Due to the reduced lactase level, lactose present in dairy products cannot be digested in the small intestine and instead is fermented by bacteria in the distal ileum and colon. The fermentative products result in symptoms of diarrhea, gas bloat, flatulence, and abdominal pain. However, in a minority of adults, high levels of lactase activity persist in adulthood. Lactase persistence is a heritable autosomal dominant condition that results in a sustained ability to digest the milk sugar lactose throughout adulthood (Olds and Sibley, 2003).
Clinical FeaturesMontgomery et al. (1991) stated that in those humans who have low lactase activity as adults, the decline occurs at approximately 3 to 5 years of age. They suggested that the developmental pattern of lactase expression is probably regulated at the level of gene transcription.
Adult-onset lactase decline appears to be a risk factor for developing osteoporosis, owing to avoidance of dairy products or interference of undigested lactose with calcium absorption (Lee and Krasinski, 1998). Elderly with both adult-onset lactase decline and atrophic gastritis or those undergoing anti-ulcer treatment may have an increased risk of low calcium absorption owing to the lack of gastric acid that facilitates calcium uptake.
Lactase and Human Evolution
• http://evolution.berkeley.edu/evolibrary/news/070401_lactose
The metabolism of galactose presentsanother problem
The solution, discovered by the Argentinian
scientist Leloir, included the discovery ofthe important class of compounds called
SUGAR NUCLEOTIDES
Lactose + H2O D-galactose + D-glucose
Galactose
1
Gal – O – P - O
Glu – O – P – O – P – O - RIBOSE - URACIL
Glu – O – P – O Glu 1-P
Gal – O – P – O – P – O - RIBOSE - URACIL
Gal 1-P
UDP-Galactose UDP-Glucose
epimerase
the reaction occurs in two steps (oxidation/reduction reactions)
oxidation
reduction
NADH + H+
NAD+
Figure 14-13
NADH + H+
NAD+
Mistake in Lehninger 4th edition and in e-book, but not in printed 5th and 6th edition
Mistake in Lehninger 4th edition, in e-book, in PowerPoint slides provided to instructors, but not in printed 6th edition
NAD+
NADH + H+
NADH + H+
NAD+
one single enzyme
no net oxidation/reduction
overall
Galactose Galactose 1-P
UDP-Glu
UDP-Gal
Glucose 1-P
Glucose 6-P
kinase
Uridyl transferase
epimerase
phosphoglucomutase
ATP ADP
Enzymes required for galactose metabolism
Glu 6-P ↔ Glu 1-P
Phosphoglucomutase
Galactosemiahigh galactose levels in blood
Possible enzyme deficiencies- galactokinase- transferase- epimerase- (mutase?)
Symptoms:large liver, icterus (jaundice), failure to thrive, and urinary excretion of
albumin and sugar; vomiting or diarrhea usually begins within a few days of milk ingestion
End of Lecture 10
October 24, 2014