Gluconeogenesis biotin

Pyruvate carboxylase is the most important of the anaplerotie reactions. It exists in the mitochondria of animal cells but not in plants, and it provides a direct link between glycolysis and the TCA cycle. The enzyme is tetrameric and contains covalently bound biotin and an Mg site on each subunit. (It is examined in greater detail in our discussion of gluconeogenesis in Chapter 23.) Pyruvate carboxylase has an absolute allosteric requirement for acetyl-CoA. Thus, when acetyl-CoA levels exceed the oxaloacetate supply, allosteric activation of pyruvate carboxylase by acetyl-CoA raises oxaloacetate levels, so that the excess acetyl-CoA can enter the TCA cycle. 

Step 1 of Figure 29.13 Carboxylation Gluconeogenesis begins with the carboxyl-afion of pyruvate to yield oxaloacetate. The reaction is catalyzed by pyruvate carboxylase and requires ATP, bicarbonate ion, and the coenzyme biotin, which acts as a carrier to transport CO2 to the enzyme active site. The mechanism is analogous to that of step 3 in fatty-acid biosynthesis (Figure 29.6), in which acetyl CoA is carboxylated to yield malonyl CoA. 

This enzyme, similar to all C02 assimilating enzymes, contains biotin for a cofactor. Oxaloacetate is released from the mitochondria into the cytoplasm to enter gluconeogenesis. In the cytoplasm, oxaloacetate converts to phosphoenolpyruvate via a reaction catalyzed by phosphoenolpyruvate carboxylase ... 

Biotin (6.24) consists of an imidazole ring fused to a tetrahydrothiophene ring with a valeric acid side chain. Biotin acts as a co-enzyme for carboxylases involved in the synthesis and catabolism of fatty acids and for branched-chain amino acids and gluconeogenesis. 

Carboxylation of pyruvate to oxaloacetate (OAA) by pyruvate carboxylase is a biotin-dependent reaction (see Figure 8.24). This reaction is important because it replenishes the citric acid cycle intermediates, and provides substrate for gluconeogenesis (see p. 116). 

Eight enzyme-catalyzed reactions are involved in the conversion of acetyl-CoA into fatty acids. The first reaction is catalyzed by acetyl-CoA carboxylase and requires ATP. This is the reaction that supplies the energy that drives the biosynthesis of fatty acids. The properties of acetyl-CoA carboxylase are similar to those of pyruvate carboxylase, which is important in the gluconeogenesis pathway (see chapter 12). Both enzymes contain the coenzyme biotin covalently linked to a lysine residue of the protein via its e-amino group. In the last section of this chapter we show that the activity of acetyl-CoA carboxylase plays an important role in the control of fatty acid biosynthesis in animals. Regulation of the first enzyme in a biosynthetic pathway is a strategy widely used in metabolism. 

Thus pyruvate carboxylase generates oxaloacetate for gluconeogenesis but also must maintain oxaloacetate levels for citric acid cycle function. For the latter reason, the activity of pyruvate carboxylase depends absolutely on the presence of acetyl CoA the biotin prosthetic group of the enzyme cannot be carboxy-lated unless acetyl CoA is bound to the enzyme. This allosteric activation by acetyl CoA ensures that more oxaloacetate is made when excess acetyl CoA is present. In this role of maintaining the level of citric acid cycle intermediates, the pyruvate carboxylase reaction is said to be anaplerotic, that is filling up. ... 

Biotin, an essential water-soluble B-complex vitamin, is the coenzyme for four human carboxylases (Fig. 12-2) These include the three mitochondrial enzymes pyruvate carboxylase, which converts pyruvate to oxaloacetate and is the initial step of gluconeogenesis propionyl-CoA carboxylase, which catabolizes several branched-chain amino acids and odd-chain fatty acids and 3-methylcrotonyl-CoA carboxylase, which is involved in the catabolism of leucine and the principally cytosolic enzyme, acetyl-CoA carboxylase, which is responsible for the... 

H Biotin Coenzyme in carboxylation reactions in gluconeogenesis and fatty acid synthesis role in regulation of cell cycle Impaired fat and carbohydrate metabolism dermatitis... 

Biotin acts to induce glucokinase, phosphofructokinase, and pyruvate kinase (key enzymes of glycolysis), phosphoenolpyruvate carboxykinase (a key enzyme of gluconeogenesis), and holocarboxylase synthetase, acting via a cell-surface receptor linked to formation of cGMP and increased activity of RNA polymerase. The activity of holocarboxylase synthetase (Section 11.2.2) falls in experimental biotin deficiency and increases with a parallel increase in... 

The activities of biotin-dependent carboxylases fall in deficiency, resulting in impaired gluconeogenesis, with accumulation of lactate, pyruvate, and alanine, and impaired lipogenesis, with accumulation of acetyl CoA, resulting in ketosis. There are also changes in the fatty acid composition of membrane lipids. A variety of abnormal organic acids are excreted by bothbiotin-deficient patients and experimental animals (as shown in Table 11.1). 

There is circumstantial evidence to support this suggestion, because the liver content of biotin is lower in infants who have died from cot death than in infants who have died from known causes. By parallel with the fatty liver and kidney syndrome, it has been suggested that a modest metabolic stress, such as a mild fever, causes a higher requirement for gluconeogenesis than can be met, resulting in acute hypoglycemia. There are rapid postmortem changes in... 

Bannister DW (1976a) The biochemistry of fatty liver and kidney syndrome. Biotin-mediated restoration of hepatic gluconeogenesis in vitro and its relationship to pyruvate carboxylase activity. Biochemical Journal 156, 167-73. 

Bannister DW (1976b) Hepatic gluconeogenesis in chicks effect of biotin on gluconeogenesis in biotin-deficiency and fatty liver and kidney syndrome. Comparative Biochemistry and Physiology B 53, 575-9. 

Arinze, J. C., and Mistry, S. P (1971). Activities of some biotin enzymes and certain aspects of gluconeogenesis during biotin deficiency. Comp. Biochem. PJjysEoL B 38,285-294. 

Carboxylation. Gluconeogenesis begins with the carboxylation of pyruvate to yield oxaloacetate. As in the third step of fatty acid synthesis (Figure 29.8), the reaction requires ATP and the coenzyme biotin, acting as a carrier of CO2. 

---