Humboldt State University ® Department of Chemistry

Richard A. Paselk

Chem 431

Biochemistry

Fall 2008

Lecture Notes: 3 December

© R. Paselk 2008
 
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Pentose Phosphate Pathway

In the non-oxidative portion of the Pentose Phosphate Pathway a series of sugar interconversions takes the RU-5-P to intermediates of other pathways: Ribose-5-P for nucleotide biosynthesis, and F-6-P and Ga-3-P for glycolysis/gluconeogenesis. All of these reactions are near equilibrium, with fluxes driven by supply and use of the three intermediates listed above.

In the first two reactions of this phase Ribulose-5-phosphate is converted either to Ribose-5-P via a 1,2-enediol intermediate, or to Xylulose-5-P via a 2,3-enediol intermediate.

These two 5-C sugars, R-5-P and Xu-5-P, are now interconverted to a 7-C sugar, Sedoheptulose-7-P, and a 3-C sugar, Glyceraldehyde-3-P. This reaction is catalyzed by Transketolase, a Thiamine pyrophosphate dependent enzyme which catalyzes the transfer of C2 units. In the first part of this reaction the TPP carbanion (ylid form) makes a nucleophilic attack on the carbonyl group of xylulose. In the resulting intermediate the C2-C3 bond is destabilized and cleavage takes place to yield the enzyme bound 2-(1,2-dihydroxyethyl)-TPP resonance stabilized carbanion:

Structural diagram of part one of the Transketolase reaction where the TPP carbanion (ylid form) makes a nucleophilic attack on the carbonyl group of xylulose. to give the 2-(1,2-dihydroxyethyl)-TPP resonance stabilized carbanion

This first part of the reaction is very similar to the first part of the Pyruvate DH catalyzed reaction in the Pyruvate DH Complex. (Ga-3-P is the leaving group instead of carbon dioxide; there is a 1,2-dihydroxyethyl instead of a 1-hydroxyethyl carbanion intermediate.) In the second part of the reaction the carbanion then attacks the aldehyde of R-5-P to give Su-7-P and regenerate the TPP catalyst:

Structural diagram of part two of the Transketolase reaction where the carbanion then attacks the aldehyde of R-5-P to give Su-7-P and regenerate the TPP catalyst

This is similar to the second part of the Pyruvate DH reaction where the hydroxyethyl group attacks the disufide of the lipoamide. (In this case, of course, the redox catalyzed by the lipoamide does not take place.)

Transaldolase catalyzes the transfer of a C3 unit. The reaction occurs via an aldol cleavage similar to that seen with aldolase: there is a schiff base intermediate formed with an active site lysine. The difference between aldolase and transaldolase is in the acceptor groups: in aldolase the acceptor is a proton, in transaldolase it is another sugar. This reaction yields a F-6-P, which can go to Glycolysis, and an E-4-P which reacts with Xu-5-P catalyzed by the same transketolase seen above. This second transketolase reaction yields F-6-P and Ga-3-P, both intermediates of Glycolysis and the end products of the Pentose-P pathway.

The interconversions of the sugars in this pathway are summarized in the flow diagram below:

flow diagram for the sugar interconversions of the pentose-P pathway

Note that the principle products of this pathway are R-5-P and NADPH. Under reductive biosynthetic conditions where R-5-P is not needed the Pentose-P pathway can be used to completely oxidize G-6-P to 6 carbon dioxide molecules with the concomitant production of 12 NADPH's. Note also that when R-5-P is needed and NADPH is not needed for reductive biosynthesis it can be made from F-6-P and Ga-3-P.

Overview of Glucose Metabolism in the Tissues: Diagram in packet [overhead]

Fat Metabolism

Fats come from two main sources: stored body fat and dietary fat. Dietary fat must first be emulsified to increase its surface area for contact with the water soluble lipases. This occurs largely in the duodenum after mixing with the bile acids, a family of cholesterol derived detergents. Triacylglycerols can then be hydrolyzed by pancreatic lipase to free fatty acids and 2-monoacylglycerol:

structural diagram of the lipase reaction

The fatty acids and monoacylglycerol are absorbed by the intestinal cells, converted to fatty acyl CoA and reassembled into triacylglycerols. The triacyl glycerols then assemble with phospholipids and lipoproteins to form chylomicrons for transport through the lymph and blood to the tissues.

When the chylomicrons reach tissue cells the triacylglycerols are again hydrolyzed by lipoprotein lipase to fatty acids which can be taken up by the peripheral tissue cells. In adipose cells the fatty acids are then converted into fatty acyl CoA's and combined into triacylglycerols for storage. Alternatively the fatty acids can be broken down for energy using the beta-oxidation pathway.

beta-oxidation of Fatty Acids

Free fatty acids are introduced into the cytosol, but -oxidation occurs in the mitosol. Two situations occur.

Short to medium length fatty acids are permeable to the mitochondrial membrane. They are activated to fatty acyl CoA derivatives in the mitochondrial matrix by Butyryl-CoA Synthetase:

structural diagram of the fattyacyl-CoA synthetase reaction

Note that two ATP equivalents are required: the phosphoanhydride and thioester bonds are of similar free energies, so a second phosphoanhydride bond is also hydrolyzed to drive the reaction to completion.

 


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Last modified 4 December 2008