Solutions to Problem Set 4

1. For mammals, fatty acids are obtained from the absorption and breakdown
of lipids obtained from the diet, and from the breakdown of the
triacylglycerides in storage tissues (adipose tissues). For germinating plant
seedlings, fatty acids are obtained from the breakdown of lipids stored in the
seed endosperm, and are also generated de novo from the sequential addition
of acetyl-CoA.
2. B, C, D, E
3. They all occur in fatty acid synthesis and result in the addition of a 2-carbon
alkane unit to a growing fatty acid chain. 2 carbon atoms.
4. The compound with the phenyl group bearing the odd-numbered fatty acid
chain would give rise to benzoic acid (benzene + COOH). In contrast,
phenylhexanoic acid would be degraded to phenylacetic acid (benzene +
CH2COOH) . These results suggested to Knoop that the fatty acids were being
metabolized from the carboxyl terminus. More importantly, the carbon
atoms were being moved two at a time, suggesting that the beta carbon atom
of the fatty acid was the likely site of oxidative degradation.
5. B, C, D. The hydrolysis of triacylglycerides yields fatty acids and glycerol,
which can be converted into glyceraldehyde-3-phosphate (GAP), a precursor
of glucose in gluconeogenesis.
6. A. First HSCoA activates the fatty acid molecule. Then the activated molecule
is oxidized by Enz(FAD). Next, it is hydrated and oxidized again, this time by
NAD
+
. An entering HSCoA then results in the thiolytic cleavage of an acetyl
CoA. (You should know why each step is important. For example, why is a
double bond introduced?) B. The first stage in fatty acid oxidation is
analogous to the conversion of succinate to fumarate; the second stage, the
conversion of fumarate to malate; the third stage, the conversion of malate to
oxaloacetate. See textbook, page 647.
7. A. Thioester bond: R-C(=O)-S-R. The thioester linkage joins the fatty acid to
HSCoA, which acts as a tag or handle by which the enzymes of the beta-
oxidation path can recognize, bind, and act on the saturated alkane chains of
the fatty acids. B. Since the change in standard free energy is approximately -
31.4 kJ/mol, it is comparable to that of the hydrolysis of ATP. The relatively
large negative value for the free energy of hydrolysis for acetyl CoA indicates
that energy must be supplied to synthesize it and that, conversely, it can
serve as an activated donor of acetyl groups. C. The carboxyl group of the
fatty acid is first activated by reaction with ATP to form an acyl-adenylate,
which contains a mixed anhydride linkage between the carboxylate and 5-
phosphate of AMP, with the release of PPi (similar to the activation of amino
acids before charging them to tRNA). Then, the acyl-CoA synthetase catalyzes
the transfer of the acyl group to the sulfhydryl group of HSCoA to form the
thioester bond and release AMP. D. The hydrolysis of PPi couples the
cleavage of a second high-energy bond to the formation of the thioester bond
to make its formation exergonic. In effect, two ATPs are used to make one
acyl-CoA.
8. A. Plants are not endothermic organisms, so they are exposed to the
environmental temperature. To maintain the membrane fluidity, they
synthesize unsaturated fatty acids. B. Because animals lack an enzyme that
can introduce double bonds beyond the C-9 position in a fatty acid, they
cannot synthesize linoleate and linolenate de novo. These unsaturated fatty
acids are precursors of a number of other needed fatty acids as well as the
eicosanoid hormones. Animals therefore rely on a diet of plants as the source
of linoleate and linolenate, which are synthesized only in plants.
9. A,b,f,g,c,d,e
10. The normal route for beta-oxidation in bacteria utilizes acyl-CoA derivatives,
which are formed from free fatty acids. To convert a primary alcohol to a free
fatty acid, two oxidative steps are needed, each requiring an electron
acceptor (Remember, oxidation reactions: alcohol aldehyde acid). In
Corynebacterium, NAD
+
-dependent dehydrogenases catalyze the sequential
conversion of a primary alcohol to a fatty acid, with the corresponding
aldehyde as an intermediate. Conversion of the free fatty acid to an acyl-CoA
derivative requires two equivalents of ATP (because ATP is converted to
AMP and PPi), as well as HSCoA (Coenzyme A). The reaction is catalyzed by
acyl-CoA synthase.
11. A. 1,3,5,8. B. 1,2,4,5,6,7.
12. The irreversible committed step of fatty acid synthesis is the formation of
malonyl-CoA from acetyl-CoA and CO2 (via HCO3
-
or bicarbonate) by acetyl-
CoA carboxylase. CO2 is fixed to acetyl-CoA to form a dicarboxylic acid at the
expense of an ATP cleavage. This facilitates the subsequent condensation
reactions with the activated acyl groups to form an acetoacyl-ACP by
releasing CO2 to help drive the reaction.
13. No. The double bond in palmitoleate is introduced by an oxidation catalyzed
by a monoxygenase that requires O2 as a co-substrate.
14. High levels of citrate signal that glucose utilization is no longer necessary and
that adequate carbon atoms are available for the synthesis of palmitoyl-CoA.
The pathways of the citric acid cycle and fatty acid synthesis do not occur in
the same intracellular compartment, thus to influence both, citrate must be
transported from the mitochondria to the cytosol.
15. A. 3 deuterium hydrogen atoms per palmitate; all located on C-16 (the
methyl end); all other two-carbon units are derived from unlabeled malonyl-
CoA. B. 7 deuteriums per palmitate; all even-numbered carbons except C-16.
16. Oxidation of fats releases metabolic water. Water is produced in the electron
transfer chain (all the NADH and FADH2 formed are going to transfer their
electrons to the electron transfer chain), when ADP and Pi are condensed as
well as in the reduction of molecular oxygen.
17. A. The net yield from the beta-oxidation of palmitate is 106 molecules of
ATP (considering that 2.5 molecules of ATP are generated for each oxidized
NADH and 1.5 for each FADH2). For the alpha-oxidation of palmitate, if one
molecule of NADH is generated for 15 of the 16 carbons of palmitate, then the
yield of ATP is 25 X 15, or 37.5. Also, the activation of the acetate molecule
requires 2 ATPs. Subsequent oxidation of acetyl-CoA through the citric acid
cycle generates 10 ATP molecules with a net yield of 8 ATPs. Thus net yeld
from the alpha-oxidation of palmitate is 37.5 + 8 = 45.5 molecules of ATP.
a. The beta-oxidation of odd-numbered fatty acids produces propionyl-
CoA, which can be converted to succinyl-CoA, a gluconeogenic
substrate. However, the alpha-oxidation of odd-numbered fatty acids
produces carbon dioxide as well as a single molecule of acetate or
acetyl-CoA, neither of which is gluconeogenic.