Introduction to metabolism

June 29, 2024
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Introduction to  metabolism. Investigation of aerobic and anaerobic oxidation of glucose.

Other ways of monosaccharides metabolism.

 

 

Carbohydrates

Foods high in carbohydrate include fruits, sweets, soft drinks, breads, pastas, beans, potatoes, bran, rice, and cereals. Carbohydrates are a common source of energy in living organisms; however, no carbohydrate is an essential nutrient in humans.

Carbohydrates are not necessary building blocks of other molecules, and the body can obtain all its energy from protein and fats.[ The brain and neurons generally cannot burn fat for energy, but use glucose or ketones. Humans can synthesize some glucose (in a set of processes known asgluconeogenesis) from specific amino acids, from the glycerol backbone in triglycerides and in some cases from fatty acids. Carbohydrate and protein contain 4 calories per gram, while fats contain 9 calories per gram. In the case of protein, this is somewhat misleading as only some amino acids are usable for fuel.

Organisms typically cannot metabolize all types of carbohydrate to yield energy. Glucose is a nearly universal and accessible source of calories. Many organisms also have the ability to metabolize other monosaccharides and Disaccharides, though glucose is preferred. In Escherichia coli, for example, the lac operon will express enzymes for the digestion of lactose when it is present, but if both lactose and glucose are present the lac operon is repressed, resulting in the glucose being used first. Polysaccharides are also common sources of energy. Many organisms can easily break down starches into glucose, however, most organisms cannot metabolize cellulose or other polysaccharides like chitin and arabinoxylans. These carbohydrates types can be metabolized by some bacteria and protists. Ruminants and termites, for example, use microorganisms to processcellulose. Even though these complex carbohydrates are not very digestible, they represent an important dietary element for humans, called dietary fiber. Fiber enhances digestion, among other benefits.

Based on the effects on risk of heart disease and obesity, the Institute of Medicine recommends that American and Canadian adults get between 45–65% of dietary energy from carbohydrates. The Food and Agriculture Organization and World Health Organization jointly recommend that national dietary guidelines set a goal of 55–75% of total energy from carbohydrates, but only 10% directly from sugars (their term for simple carbohydrates).

 

Functions of Carbohydrate

All animals derive the major portion of their food calories from the different types of Carbohydrates in their diets. Most of the energy for the metabolic activities of the cell in all organisms is derived from the oxidation of Carbohydrate. Important functions of Carbohydrate are that of storing food, acting as a framework in body, performs are listed below.

 

Carbohydrate functions as Bio Fuel

Carbohydrate functions as an energy source of the body and acts as Bio fuel.Step wise details for the process of production of energy are discussed below.

·                     Polysaccharides such as starch and glycogen are first hydrolyzed by enzymes to Glucose.

·                     Glucose is the transported from one cell to another by blood in case of animals and cell sap in case of plants.

·                     Glucose is then oxidized to produce carbon dioxide and water.

·                     Energy is released in this process which is used for functioning of the cells.

Carbohydrate functions as Primary Source of Energy

The process of production of energy by carbohydrates is described in above steps. Now it is important to note, that fats and proteins can also be burned to provide energy but carbohydrate functions as primary source of energy. Fats are only burned if there is non availability of carbohydrates. When fat is burned in absence of carbohydrates, toxic compounds like called ketone bodies are produced. Accumulation of these ketone bodies over long period causes a condition called Ketosis. In this condition blood becomes unable to carry oxygen properly and this can be fatal. Thus, one of important function of carbohydrate is help burn fat properly.

Carbohydrate functions as storage food

Different forms of Carbohydrate are stored in living organism as storage food.

·                     Polysaccharide starch acts as storage food for plants.

·                     Glycogen stored in liver and muscles acts as storage food for animals.

·                     Inulin acts as storage food of dahlias, onion and garlic.

Thus carbohydrate performs the function of storing food.

Carbohydrate functions as framework in body

Different Carbohydrates especially Polysaccharides act as framework in living organism.

·                     Cellulose forms cell wall of plant cell along with hemicelluloses and Pectin

·                     Chitin   forms cell wall of fungal cell and exoskeleton of arthropods

·                     Peptidoglycan forms cell wall of bacteria and cyanobacteria.

Thus carbohydrates function as contributing material to the cellular structure.

Carbohydrate functions as Anticoagulant

Heparin is a polysaccharide (carbohydrate) which acts as anticoagulant and prevents intravascular clotting.

Carbohydrate functions as Antigen

Many antigens are glycoprotein (which contains oligosaccharide) iature and give immunological properties to the blood.

Carbohydrate functions as Hormone

Many Hormones like FSH (Follicular Stimulating Hormone which takes part in ovulation in females) and LH (Leutinizing Hormone) are glycoprotein and help in reproductive processes.

Carbohydrates provide raw material for industry

Carbohydrates are an important component of many industries like textile, paper, lacquers and breweries.

Other Functions

Agar is polysaccharide used in culture media, laxative and food.

Cellulose acts as roughage of food. It stimulates peristalsis movement and secretion of digestive enzymes.

Hyaluronic acid found in between joints acts as synovial fluid and provides frictionless movement.

 

 

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Carbohydrates have the general molecular formula CH2O, and thus were once thought to represent “hydrated carbon”. However, the arrangement of atoms in carbohydrates has little to do with water molecules.

http://www.youtube.com/watch?v=p-lFJVOkFwk

Starch and cellulose are two common carbohydrates. Both are macromolecules with molecular weights in the hundreds of thousands. Both are polymers (hence “polysaccharides“); that is, each is built from repeating units, monomers, much as a chain is built from its links.

The monomers of both starch and cellulose are the same: units of the sugar glucose.

Sugars

Monosaccharides

hexosesThree common sugars share the same molecular formula: C6H12O6. Because of their six carbon atoms, each is a hexose.

They are:

·                    glucose, “blood sugar”, the immediate source of energy for cellular respiration

·                    galactose, a sugar in milk (and yogurt), and

·                    fructose, a sugar found in honey.

Although all three share the same molecular formula (C6H12O6), the arrangement of atoms differs in each case. Substances such as these three, which have identical molecular formulas but different structural formulas, are known as structural isomers.

Glucose, galactose, and fructose are “single” sugars or monosaccharides. Two monosaccharides can be linked together to form a “double” sugar or disaccharide.

Disaccharides

Three common disaccharides:

·                    sucrose — common table sugar = glucose + fructose

·                    lactose — major sugar in milk = glucose + galactose

·                    maltose — product of starch digestion = glucose + glucose

Although the process of linking the two monomers is rather complex, the end result in each case is the loss of a hydrogen atom (H) from one of the monosaccharides and a hydroxyl group (OH) from the other. The resulting linkage between the sugars is called a glycosidic bond. The molecular formula of each of these disaccharides is

C12H22O11 = 2 C6H12O6 − H2O

All sugars are very soluble in water because of their many hydroxyl groups. Although not as concentrated a fuel as fats, sugars are the most important source of energy for many cells.

Carbohydrates provide the bulk of the calories (4 kcal/gram) in most diets, and starches provide the bulk of that. Starches are polysaccharides.

Polysaccharides

Starches

starch

Starches are polymers of glucose. Two types are found:

·                    amylose consists of linear, unbranched chains of several hundred glucose residues (units). The glucose residues are linked by a glycosidic bond between their #1 and #4 carbon atoms.

·                    amylopectin differs from amylose in being highly branched. At approximately every thirtieth residue along the chain, a short side chain is attached by a glycosidic bond to the #6 carbon atom (the carbon above the ring). The total number of glucose residues in a molecule of amylopectin is several thousand.

potato

Starches are insoluble in water and thus can serve as storage depots of glucose. Plants convert excess glucose into starch for storage. The image shows starch grains (lightly stained with iodine) in the cells of the white potato. Rice, wheat, and corn are also major sources of starch in the human diet.


amylase

Before starches can enter (or leave) cells, they must be digested. The hydrolysis of starch is done by amylases. With the aid of an amylase (such as pancreatic amylase), water molecules enter at the 1 -> 4 linkages, breaking the chain and eventually producing a mixture of glucose and maltose. A different amylase is needed to break the 1 -> 6 bonds of amylopectin.

http://www.youtube.com/watch?v=AEsQxzeAry8

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Glycogen

Animals store excess glucose by polymerizing it to form glycogen. The structure of glycogen is similar to that of amylopectin, although the branches in glycogen tend to be shorter and more frequent.

Glycogen is broken back down into glucose when energy is needed (a process called glycogenolysis).

http://www.youtube.com/watch?v=D2RFc1D_Iv0&feature=related

 

In glycogenolysis,

·                    phosphate groups — not water — break the 1 -> 4 linkages

·                    the phosphate group must then be removed so that glucose can leave the cell.

The liver and skeletal muscle are major depots of glycogen.

There is some evidence that intense exercise and a high-carbohydrate diet (“carbo-loading”) can increase the reserves of glycogen in the muscles and thus may help marathoners work their muscles somewhat longer and harder than otherwise. But for most of us, carbo loading leads to increased deposits of fat.

Cellulose

cellulose

Cellulose is probably the single most abundant organic molecule in the biosphere. It is the major structural material of which plants are made. Wood is largely cellulose while cotton and paper are almost pure cellulose.

Like starch, cellulose is a polysaccharide with glucose as its monomer. However, cellulose differs profoundly from starch in its properties.

Preston2

·                    Because of the orientation of the glycosidic bonds linking the glucose residues, the rings of glucose are arranged in a flip-flop manner. This produces a long, straight, rigid molecule.

·                    There are no side chains in cellulose as there are in starch. The absence of side chains allows these linear molecules to lie close together.

·                    Because of the many -OH groups, as well as the oxygen atom in the ring, there are many opportunities for hydrogen bonds to form between adjacent chains.

The result is a series of stiff, elongated fibrils — the perfect material for building the cell walls of plants.

This electron micrograph (courtesy of R. D. Preston) shows the cellulose fibrils in the cell wall of a green alga. These long, rigid fibrils are a clear reflection of the nature of the cellulose molecules of which they are composed.

Carbohydrates and Metabolism Chart – Carb Chart

Carb Chart - Digestion & Metabolism

http://www.swet.nl/images/2_01.jpghttp://www.swet.nl/images/2_02.jpghttp://www.swet.nl/images/2_03.jpg

 

 

Digestion of Dietary Carbohydrates

Dietary carbohydrate from which humans gain energy enter the body in complex forms, such as disaccharides and the polymers starch (amylose and amylopectin) and glycogen. The polymer cellulose is also consumed but not digested. The first step in the metabolism of digestible carbohydrate is the conversion of the higher polymers to simpler, soluble forms that can be transported across the intestinal wall and delivered to the tissues. The breakdown of polymeric sugars begins in the mouth. Saliva has a slightly acidic pH of 6.8 and contains lingual amylase that begins the digestion of carbohydrates. The action of lingual amylase is limited to the area of the mouth and the esophagus; it is virtually inactivated by the much stronger acid pH of the stomach. Once the food has arrived in the stomach, acid hydrolysis contributes to its degradation; specific gastric proteases and lipases aid this process for proteins and fats, respectively. The mixture of gastric secretions, saliva, and food, known collectively as chyme, moves to the small intestine.

The main polymeric-carbohydrate digesting enzyme of the small intestine is -amylase. This enzyme is secreted by the pancreas and has the same activity as salivary amylase, producing disaccharides and trisaccharides. The latter are converted to monosaccharides by intestinal saccharidases, including maltases that hydrolyze di- and trisaccharides, and the more specific disaccharidases, sucrase, lactase, and trehalase. The net result is the almost complete conversion of digestible carbohydrate to its constituent monosaccharides. The resultant glucose and other simple carbohydrates are transported across the intestinal wall to the hepatic portal vein and then to liver parenchymal cells and other tissues. There they are converted to fatty acids, amino acids, and glycogen, or else oxidized by the various catabolic pathways of cells.

Oxidation of glucose is known as glycolysis.Glucose is oxidized to either lactate or pyruvate. Under aerobic conditions, the dominant product in most tissues is pyruvate and the pathway is known as aerobic glycolysis. When oxygen is depleted, as for instance during prolonged vigorous exercise, the dominant glycolytic product in many tissues is lactate and the process is known as anaerobic glycolysis.
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The Energy Derived from Glucose Oxidation

Aerobic glycolysis of glucose to pyruvate, requires two equivalents of ATP to activate the process, with the subsequent production of four equivalents of ATP and two equivalents of NADH. Thus, conversion of one mole of glucose to two moles of pyruvate is accompanied by the net production of two moles each of ATP and NADH.

Glucose + 2 ADP + 2 NAD+ + 2 Pi —–> 2 Pyruvate + 2 ATP + 2 NADH + 2 H+

The NADH generated during glycolysis is used to fuel mitochondrial ATP synthesis via oxidative phosphorylation, producing either two or three equivalents of ATP depending upon whether the glycerol phosphate shuttle or the malate-aspartate shuttle is used to transport the electrons from cytoplasmic NADH into the mitochondria. The net yield from the oxidation of 1 mole of glucose to 2 moles of pyruvate is, therefore, either 6 or 8 moles of ATP. Complete oxidation of the 2 moles of pyruvate, through the TCA cycle, yeilds an additional 30 moles of ATP; the total yield, therefore being either 36 or 38 moles of ATP from the complete oxidation of 1 mole of glucose to CO2 and H2O.
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The Individual Reactions of Glycolysis

The pathway of glycolysis can be seen as consisting of 2 separate phases. The first is the chemical priming phase requiring energy in the form of ATP, and the second is considered the energy-yielding phase. In the first phase, 2 equivalents of ATP are used to convert glucose to fructose 1,6-bisphosphate (F1,6BP). In the second phase F1,6BP is degraded to pyruvate, with the production of 4 equivalents of ATP and 2 equivalents of NADH.

http://www.youtube.com/watch?v=6JGXayUyNVw&feature=related

http://www.youtube.com/watch?v=nKgUBsC4Oyo&feature=related

 

http://web.indstate.edu/thcme/mwking/glycolysispathway_a.gif

http://web.indstate.edu/thcme/mwking/glycolysispathway_b.gif

Pathway of glycolysis from glucose to pyruvate. Substrates and products are in blue, enzymes are in green. The two high energy intermediates whose oxidations are coupled to ATP synthesis are shown in red (1,3-bisphosphoglycerate and phosphoenolpyruvate).

http://www.youtube.com/watch?v=PQMsJSme780&feature=related

 

The Hexokinase Reaction:

The ATP-dependent phosphorylation of glucose to form glucose 6-phosphate (G6P)is the first reaction of glycolysis, and is catalyzed by tissue-specific isoenzymes known as hexokinases. The phosphorylation accomplishes two goals: First, the hexokinase reaction converts nonionic glucose into an anion that is trapped in the cell, since cells lack transport systems for phosphorylated sugars. Second, the otherwise biologically inert glucose becomes activated into a labile form capable of being further metabolized.

Four mammalian isozymes of hexokinase are known (Types I – IV), with the Type IV isozyme often referred to as glucokinase. Glucokinase is the form of the enzyme found in hepatocytes. The high Km of glucokinase for glucose means that this enzyme is saturated only at very high concentrations of substrate.

 

http://web.indstate.edu/thcme/mwking/hexokinasereaction.gif

Comparison of the activities of hexokinase and glucokinase. The Km for hexokinase is significantly lower (0.1mM) than that of glucokinase (10mM). This difference ensures that non-hepatic tissues (which contain hexokinase) rapidly and efficiently trap blood glucose within their cells by converting it to glucose-6-phosphate. One major function of the liver is to deliver glucose to the blood and this in ensured by having a glucose phosphorylating enzyme (glucokinase) whose Km for glucose is sufficiently higher that the normal circulating concentration of glucose (5mM).

 

This feature of hepatic glucokinase allows the liver to buffer blood glucose. After meals, when postprandial blood glucose levels are high, liver glucokinase is significantly active, which causes the liver preferentially to trap and to store circulating glucose. When blood glucose falls to very low levels, tissues such as liver and kidney, which contain glucokinases but are not highly dependent on glucose, do not continue to use the meager glucose supplies that remain available. At the same time, tissues such as the brain, which are critically dependent on glucose, continue to scavenge blood glucose using their low Km hexokinases, and as a consequence their viability is protected. Under various conditions of glucose deficiency, such as long periods between meals, the liver is stimulated to supply the blood with glucose through the pathway of gluconeogenesis. The levels of glucose produced during gluconeogenesis are insufficient to activate glucokinase, allowing the glucose to pass out of hepatocytes and into the blood.

The regulation of hexokinase and glucokinase activities is also different. Hexokinases I, II, and III are allosterically inhibited by product (G6P) accumulation, whereas glucokinases are not. The latter further insures liver accumulation of glucose stores during times of glucose excess, while favoring peripheral glucose utilization when glucose is required to supply energy to peripheral tissues.

Phosphohexose Isomerase:

The second reaction of glycolysis is an isomerization, in which G6P is converted to fructose 6-phosphate (F6P). The enzyme catalyzing this reaction is phosphohexose isomerase (also known as phosphoglucose isomerase). The reaction is freely reversible at normal cellular concentrations of the two hexose phosphates and thus catalyzes this interconversion during glycolytic carbon flow and during gluconeogenesis.

6-Phosphofructo-1-Kinase (Phosphofructokinase-1, PFK-1):

The next reaction of glycolysis involves the utilization of a second ATP to convert F6P to fructose 1,6-bisphosphate (F1,6BP). This reaction is catalyzed by 6-phosphofructo-1-kinase, better known as phosphofructokinase-1 or PFK-1. This reaction is not readily reversible because of its large positive free energy (G0′ = +5.4 kcal/mol) in the reverse direction. Nevertheless, fructose units readily flow in the reverse (gluconeogenic) direction because of the ubiquitous presence of the hydrolytic enzyme, fructose-1,6-bisphosphatase (F-1,6-BPase).

The presence of these two enzymes in the same cell compartment provides an example of a metabolic futile cycle, which if unregulated would rapidly deplete cell energy stores. However, the activity of these two enzymes is so highly regulated that PFK-1 is considered to be the rate-limiting enzyme of glycolysis and F-1,6-BPase is considered to be the rate-limiting enzyme in gluconeogenesis.

Aldolase:

Aldolase catalyses the hydrolysis of F1,6BP into two 3-carbon products: dihydroxyacetone phosphate (DHAP) and glyceraldehyde 3-phosphate (G3P). The aldolase reaction proceeds readily in the reverse direction, being utilized for both glycolysis and gluconeogenesis.

Triose Phosphate Isomerase: \

The two products of the aldolase reaction equilibrate readily in a reaction catalyzed by triose phosphate isomerase. Succeeding reactions of glycolysis utilize G3P as a substrate; thus, the aldolase reaction is pulled in the glycolytic direction by mass action principals.

Glyceraldehyde-3-Phosphate Dehydrogenase:

The second phase of glucose catabolism features the energy-yielding glycolytic reactions that produce ATP and NADH. In the first of these reactions, glyceraldehyde-3-P dehydrogenase (G3PDH) catalyzes the NAD+-dependent oxidation of G3P to 1,3-bisphosphoglycerate (1,3BPG) and NADH. The G3PDH reaction is reversible, and the same enzyme catalyzes the reverse reaction during gluconeogenesis.

Phosphoglycerate Kinase:

The high-energy phosphate of 1,3-BPG is used to form ATP and 3-phosphoglycerate (3PG) by the enzyme phosphoglycerate kinase. Note that this is the only reaction of glycolysis or gluconeogenesis that involves ATP and yet is reversible under normal cell conditions. Associated with the phosphoglycerate kinase pathway is an important reaction of erythrocytes, the formation of 2,3-bisphosphoglycerate, 2,3BPG (see Figure below) by the enzyme bisphosphoglycerate mutase. 2,3BPG is an important regulator of hemoglobin’s affinity for oxygen. Note that 2,3-bisphosphoglycerate phosphatase degrades 2,3BPG to 3-phosphoglycerate, a normal intermediate of glycolysis. The 2,3BPG shunt thus operates with the expenditure of 1 equivalent of ATP per triose passed through the shunt. The process is not reversible under physiological conditions.

 

http://web.indstate.edu/thcme/mwking/23bpg.gif

The pathway for 2,3-bisphosphoglycerate (2,3-BPG) synthesis within erythrocytes. Synthesis of 2,3-BPG represents a major reaction pathway for the consumption of glucose in erythrocytes. The synthesis of 2,3-BPG in erythrocytes is critical for controlling hemoglobin affinity for oxygen. Note that when glucose is oxidized by this pathway the erythrocyte loses the ability to gain 2 moles of ATP from glycolytic oxidation of 1,3-BPG to 3-phosphoglycerate via the phosphoglycerate kinase reaction.

Phosphoglycerate Mutase and Enolase:

The remaining reactions of glycolysis are aimed at converting the relatively low energy phosphoacyl-ester of 3PG to a high-energy form and harvesting the phosphate as ATP. The 3PG is first converted to 2PG by phosphoglycerate mutase and the 2PG conversion to phosphoenoylpyruvate (PEP) is catalyzed by enolase

Pyruvate Kinase:

The final reaction of aerobic glycolysis is catalyzed by the highly regulated enzyme pyruvate kinase (PK). In this strongly exergonic reaction, the high-energy phosphate of PEP is conserved as ATP. The loss of phosphate by PEP leads to the production of pyruvate in an unstable enol form, which spontaneously tautomerizes to the more stable, keto form of pyruvate. This reaction contributes a large proportion of the free energy of hydrolysis of PEP.

Anaerobic Glycolysis

 

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Under aerobic conditions, pyruvate in most cells is further metabolized via the TCA cycle. Under anaerobic conditions and in erythrocytes under aerobic conditions, pyruvate is converted to lactate by the enzyme lactate dehydrogenase (LDH), and the lactate is transported out of the cell into the circulation. The conversion of pyruvate to lactate, under anaerobic conditions, provides the cell with a mechanism for the oxidation of NADH (produced during the G3PDH reaction) to NAD+; which occurs during the LDH catalyzed reaction. This reduction is required since NAD+ is a necessary substrate for G3PDH, without which glycolysis will cease. Normally, during aerobic glycolysis the electrons of cytoplasmic NADH are transferred to mitochondrial carriers of the oxidative phosphorylation pathway generating a continuous pool of cytoplasmic NAD+.

Aerobic glycolysis generates substantially more ATP per mole of glucose oxidized than does anaerobic glycolysis. The utility of anaerobic glycolysis, to a muscle cell when it needs large amounts of energy, stems from the fact that the rate of ATP production from glycolysis is approximately 100X faster than from oxidative phosphorylation. During exertion muscle cells do not need to energize anabolic reaction pathways. The requirement is to generate the maximum amount of ATP, for muscle contraction, in the shortest time frame. This is why muscle cells derive almost all of the ATP consumed during exertion from anaerobic glycolysis.

The reactions catalyzed by hexokinase, PFK-1 and PK all proceed with a relatively large free energy decrease. These nonequilibrium reactions of glycolysis would be ideal candidates for regulation of the flux through glycolysis. Indeed, in vitro studies have shown all three enzymes to be allosterically controlled.

Regulation of hexokinase, however, is not the major control point in glycolysis. This is due to the fact that large amounts of G6P are derived from the breakdown of glycogen (the predominant mechanism of carbohydrate entry into glycolysis in skeletal muscle) and, therefore, the hexokinase reaction is not necessary. Regulation of PK is important for reversing glycolysis when ATP is high in order to activate gluconeogenesis. As such this enzyme catalyzed reaction is not a major control point in glycolysis. The rate limiting step in glycolysis is the reaction catalyzed by PFK-1.

PFK-1 is a tetrameric enzyme that exist in two conformational states termed R and T that are in equilibrium. ATP is both a substrate and an allosteric inhibitor of PFK-1. Each subunit has two ATP binding sites, a substrate site and an inhibitor site. The substrate site binds ATP equally well when the tetramer is in either conformation. The inhibitor site binds ATP essentially only when the enzyme is in the T state. F6P is the other substrate for PFK-1 and it also binds preferentially to the R state enzyme. At high concentrations of ATP, the inhibitor site becomes occupied and shifting the equilibrium of PFK-1 comformation to that of the T state decreasing PFK-1’s ability to bind F6P. The inhibition of PFK-1 by ATP is overcome by AMP which binds to the R state of the enzyme and, therefore, stabilizes the conformation of the enzyme capable of binding F6P. The most important allosteric regulator of both glycolysis and gluconeogenesis is fructose 2,6-bisphosphate, F2,6BP, which is not an intermediate in glycolysis or in gluconeogenesis.

 

http://web.indstate.edu/thcme/mwking/fructose26bisphosphate.gif

Regulation of glycolysis and gluconeogenesis by fructose 2,6-bisphosphate (F2,6BP). The major sites for regulation of glycolysis and gluconeogenesis are the phosphofructokinase-1 (PFK-1) and fructose-1,6-bisphosphatase (F-1,6-BPase) catalyzed reactions. PFK-2 is the kinase activity and F-2,6-BPase is the phosphatase activity of the bi-functional regulatory enzyme, phosphofructokinase-2/fructose-2,6-bisphosphatase. PKA is cAMP-dependent protein kinase which phosphorylates PFK-2/F-2,6-BPase turning on the phosphatase activity. (+ve) and (-ve) refer to positive and negative activities, respectively.

 

The synthesis of F2,6BP is catalyzed by the bifunctional enzyme phosphofructokinase-2/fructose-2,6-bisphosphatase (PFK-2/F-2,6-BPase). In the nonphosphorylated form the enzyme is known as PFK-2 and serves to catalyze the synthesis of F2,6BP by phosphorylating fructose 6-phosphate. The result is that the activity of PFK-1 is greatly stimulated and the activity of F-1,6-BPase is greatly inhibited.

Under conditions where PFK-2 is active, fructose flow through the PFK-1/F-1,6-BPase reactions takes place in the glycolytic direction, with a net production of F1,6BP. When the bifunctional enzyme is phosphorylated it no longer exhibits kinase activity, but a new active site hydrolyzes F2,6BP to F6P and inorganic phosphate. The metabolic result of the phosphorylation of the bifunctional enzyme is that allosteric stimulation of PFK-1 ceases, allosteric inhibition of F-1,6-BPase is eliminated, and net flow of fructose through these two enzymes is gluconeogenic, producing F6P and eventually glucose.

The interconversion of the bifunctional enzyme is catalyzed by cAMP-dependent protein kinase (PKA), which in turn is regulated by circulating peptide hormones. When blood glucose levels drop, pancreatic insulin production falls, glucagon secretion is stimulated, and circulating glucagon is highly increased. Hormones such as glucagon bind to plasma membrane receptors on liver cells, activating membrane-localized adenylate cyclase leading to an increase in the conversion of ATP to cAMP (see diagram below). cAMP binds to the regulatory subunits of PKA, leading to release and activation of the catalytic subunits. PKA phosphorylates numerous enzymes, including the bifunctional PFK-2/F-2,6-BPase. Under these conditions the liver stops consuming glucose and becomes metabolically gluconeogenic, producing glucose to reestablish normoglycemia.

 

http://web.indstate.edu/thcme/mwking/pkacamp.jpg

Representative pathway for the activation of cAMP-dependent protein kinase (PKA). In this example glucagon binds to its’ cell-surface receptor, thereby activating the receptor. Activation of the receptor is coupled to the activation of a receptor-coupled G-protein (GTP-binding and hydrolyzing protein) composed of 3 subunits. Upon activation the alpha subunit dissociates and binds to and activates adenylate cyclase. Adenylate cylcase then converts ATP to cyclic-AMP (cAMP). The cAMP thus produced then binds to the regulatory subunits of PKA leading to dissociation of the associated catalytic subunits. The catalytic subunits are inactive until dissociated from the regulatory subunits. Once released the catalytic subunits of PKA phosphorylate numerous substrate using ATP as the phosphate donor.

 

Regulation of glycolysis also occurs at the step catalyzed by pyruvate kinase, (PK). The liver enzyme has been most studied in vitro. This enzyme is inhibited by ATP and acetyl-CoA and is activated by F1,6BP. The inhibition of PK by ATP is similar to the effect of ATP on PFK-1. The binding of ATP to the inhibitor site reduces its affinity for PEP. The liver enzyme is also controlled at the level of synthesis. Increased carbohydrate ingestion induces the synthesis of PK resulting in elevated cellular levels of the enzyme.

A number of PK isozymes have been described. The liver isozyme (L-type), characteristic of a gluconeogenic tissue, is regulated via phosphorylation by PKA, whereas the M-type isozyme found in brain, muscle, and other glucose requiring tissue is unaffected by PKA. As a consequence of these differences, blood glucose levels and associated hormones can regulate the balance of liver gluconeogenesis and glycolysis while muscle metabolism remains unaffected.

In erythrocytes, the fetal PK isozyme has much greater activity than the adult isozyme; as a result, fetal erythrocytes have comparatively low concentrations of glycolytic intermediates. Because of the low steady-state concentration of fetal 1,3BPG, the 2,3BPG shunt (see diagram above) is greatly reduced in fetal cells and little 2,3BPG is formed. Since 2,3BPG is a negative effector of hemoglobin affinity for oxygen, fetal erythrocytes have a higher oxygen affinity than maternal erythrocytes. Therefore, transfer of oxygen from maternal hemoglobin to fetal hemoglobin is favored, assuring the fetal oxygen supply. In the newborn, an erythrocyte isozyme of the M-type with comparatively low PK activity displaces the fetal type, resulting in an accumulation of glycolytic intermediates. The increased 1,3BPG levels activate the 2,3BPG shunt, producing 2,3BPG needed to regulate oxygen binding to hemoglobin.

Genetic diseases of adult erythrocyte PK are known in which the kinase is virtually inactive. The erythrocytes of affected individuals have a greatly reduced capacity to make ATP and thus do not have sufficient ATP to perform activities such as ion pumping and maintaining osmotic balance. These erythrocytes have a short half-life, lyse readily, and are responsible for some cases of hereditary hemolytic anemia.

The liver PK isozyme is regulated by phosphorylation, allosteric effectors, and modulation of gene expression. The major allosteric effectors are F1,6BP, which stimulates PK activity by decreasing its Km(app) for PEP, and for the negative effector, ATP. Expression of the liver PK gene is strongly influenced by the quantity of carbohydrate in the diet, with high-carbohydrate diets inducing up to 10-fold increases in PK concentration as compared to low carbohydrate diets. Liver PK is phosphorylated and inhibited by PKA, and thus it is under hormonal control similar to that described earlier for PFK-2.

Muscle PK (M-type) is not regulated by the same mechanisms as the liver enzyme. Extracellular conditions that lead to the phosphorylation and inhibition of liver PK, such as low blood glucose and high levels of circulating glucagon, do not inhibit the muscle enzyme. The result of this differential regulation is that hormones such as glucagon and epinephrine favor liver gluconeogenesis by inhibiting liver glycolysis, while at the same time, muscle glycolysis can proceed in accord with needs directed by intracellular conditions.

Metabolic Fates of Pyruvate

Pyruvate is the branch point molecule of glycolysis. The ultimate fate of pyruvate depends on the oxidation state of the cell. In the reaction catalyzed by G3PDH a molecule of NAD+ is reduced to NADH. In order to maintain the re-dox state of the cell, this NADH must be re-oxidized to NAD+. During aerobic glycolysis this occurs in the mitochondrial electron transport chain generating ATP. Thus, during aerobic glycolysis ATP is generated from oxidation of glucose directly at the PGK and PK reactions as well as indirectly by re-oxidation of NADH in the oxidative phosphorylation pathway. Additional NADH molecules are generated during the complete aerobic oxidation of pyruvate in the TCA cycle. Pyruvate enters the TCA cycle in the form of acetyl-CoA which is the product of the pyruvate dehydrogenase reaction. The fate of pyruvate during anaerobic glycolysis is reduction to lactate.

Lactate Metabolism

During anaerobic glycolysis, that period of time when glycolysis is proceeding at a high rate (or in anaerobic organisms), the oxidation of NADH occurs through the reduction of an organic substrate. Erythrocytes and skeletal muscle (under conditions of exertion) derive all of their ATP needs through anaerobic glycolysis. The large quantity of NADH produced is oxidized by reducing pyruvate to lactate. This reaction is carried out by lactate dehydrogenase, (LDH). The lactate produced during anaerobic glycolysis diffuses from the tissues and is transproted to highly aerobic tissues such as cardiac muscle and liver. The lactate is then oxidized to pyruvate in these cells by LDH and the pyruvate is further oxidized in the TCA cycle. If the energy level in these cells is high the carbons of pyruvate will be diverted back to glucose via the gluconeogenesis pathway.

Mammalian cells contain two distinct types of LDH subunits, termed M and H. Combinations of these different subunits generates LDH isozymes with different characteristics. The H type subunit predominates in aerobic tissues such as heart muscle (as the H4 tetramer) while the M subunit predominates in anaerobic tissues such as skeletal muscle as the M4 tetramer). H4 LDH has a low Km for pyruvate and also is inhibited by high levels of pyruvate. The M4 LDH enzyme has a high Km for pyruvate and is not inhibited by pyruvate. This suggsts that the H-type LDH is utilized for oxidizing lactate to pyruvate and the M-type the reverse.
Ethanol Metabolism

Animal cells (primarily hepatocytes) contain the cytosolic enzyme alcohol dehydrogenase (ADH) which oxidizes ethanol to acetaldehyde. Acetaldehyde then enters the mitochondria where it is oxidized to acetate by acetaldehyde dehydrogenase (AcDH).

 

http://web.indstate.edu/thcme/mwking/alcoholmetabolism.jpg

 

Acetaldehyde forms adducts with proteins, nucleic acids and other compounds, the results of which are the toxic side effects (the hangover) that are associated with alcohol consumption. The ADH and AcDH catalyzed reactions also leads to the reduction of NAD+ to NADH. The metabolic effects of ethanol intoxication stem from the actions of ADH and AcDH and the resultant cellular imbalance in the NADH/NAD+. The NADH produced in the cytosol by ADH must be reduced back to NAD+ via either the malate-aspartate shuttle or the glycerol-phosphate shuttle. Thus, the ability of an individual to metabolize ethanol is dependent upon the capacity of hepatocytes to carry out eother of these 2 shuttles, which in turn is affected by the rate of the TCA cycle in the mitochondria whose rate of function is being impacted by the NADH produced by the AcDH reaction. The reduction in NAD+ impairs the flux of glucose through glycolysis at the glyceraldehyde-3-phosphate dehydrogenase reaction, thereby limiting energy production. Additionally, there is an increased rate of hepatic lactate production due to the effect of increased NADH on direction of the hepatic lactate dehydrogenase (LDH) reaction. This reverseral of the LDH reaction in hepatocytes diverts pyruvate from gluconeogenesis leading to a reduction in the capacity of the liver to deliver glucose to the blood.

In addition to the negative effects of the altered NADH/NAD+ ratio on hepatic gluconeogenesis, fatty acid oxidation is also reduced as this process requires NAD+ as a cofactor. In fact the opposite is true, fatty acid synthesis is increased and there is an increase in triacylglyceride production by the liver. In the mitocondria, the production of acetate from acetaldehyde leads to increased levels of acetyl-CoA. Since the increased generation of NADH also reduces the activity of the TCA cycle, the acetyl-CoA is diverted to fatty acid synthesis. The reduction in cytosolic NAD+ leads to reduced activity of glycerol-3-phosphate dehydrogenase (in the glcerol 3-phosphate to DHAP direction) resulting in increased levels of glycerol 3-phosphate which is the backbone for the synthesis of the triacylglycerides. Both of these two events lead to fatty acid deposition in the liver leading to fatty liver syndrome.

 

Regulation of Blood Glucose Levels

If for no other reason, it is because of the demands of the brain for oxidizable glucose that the human body exquisitely regulates the level of glucose circulating in the blood. This level is maintained in the range of 5mM.

Nearly all carbohydrates ingested in the diet are converted to glucose following transport to the liver. Catabolism of dietary or cellular proteins generates carbon atoms that can be utilized for glucose synthesis via gluconeogenesis. Additionally, other tissues besides the liver that incompletely oxidize glucose (predominantly skeletal muscle and erythrocytes) provide lactate that can be converted to glucose via gluconeogenesis.

Maintenance of blood glucose homeostasis is of paramount importance to the survival of the human organism. The predominant tissue responding to signals that indicate reduced or elevated blood glucose levels is the liver. Indeed, one of the most important functions of the liver is to produce glucose for the circulation. Both elevated and reduced levels of blood glucose trigger hormonal responses to initiate pathways designed to restore glucose homeostasis. Low blood glucose triggers release of glucagon from pancreatic -cells. High blood glucose triggers release of insulin from pancreatic -cells. Additional signals, ACTH and growth hormone, released from the pituitary act to increase blood glucose by inhibiting uptake by extrahepatic tissues. Glucocorticoids also act to increase blood glucose levels by inhibiting glucose uptake. Cortisol, the major glucocorticoid released from the adrenal cortex, is secreted in response to the increase in circulating ACTH. The adrenal medullary hormone, epinephrine, stimulates production of glucose by activating glycogenolysis in response to stressful stimuli.

Glucagon binding to its’ receptors on the surface of liver cells triggers an increase in cAMP production leading to an increased rate of glycogenolysis by activating glycogen phosphorylase via the PKA-mediated cascade. This is the same response hepatocytes have to epinephrine release. The resultant increased levels of G6P in hepatocytes is hydrolyzed to free glucose, by glucose-6-phosphatase, which then diffuses to the blood. The glucose enters extrahepatic cells where it is re-phosphorylated by hexokinase. Since muscle and brain cells lack glucose-6-phosphatase, the glucose-6-phosphate product of hexokinase is retained and oxidized by these tissues.

In opposition to the cellular responses to glucagon (and epinephrine on hepatocytes), insulin stimulates extrahepatic uptake of glucose from the blood and inhibits glycogenolysis in extrahepatic cells and conversely stimulates glycogen synthesis. As the glucose enters hepatocytes it binds to and inhibits glycogen phosphorylase activity. The binding of free glucose stimulates the de-phosphorylation of phosphorylase thereby, inactivating it. Why is it that the glucose that enters hepatocytes is not immediately phosphorylated and oxidized? Liver cells contain an isoform of hexokinase called glucokinase. Glucokinase has a much lower affinity for glucose than does hexokinase. Therefore, it is not fully active at the physiological ranges of blood glucose. Additionally, glucokinase is not inhibited by its product G6P, whereas, hexokinase is inhibited by G6P.

One major response of non-hepatic tissues to insulin is the recruitment, to the cell surface, of glucose transporter complexes. Glucose transporters comprise a family of five members, GLUT-1 to GLUT-5. GLUT-1 is ubiquitously distributed in various tissues. GLUT-2 is found primarily in intestine, kidney and liver. GLUT-3 is also found in the intestine and GLUT-5 in the brain and testis. GLUT-5 is also the major glucose transporter present in the membrane of the endoplasmic reticulum (ER) and serves the function of transporting glucose to the cytosol following its‘ dephosphorylation by the ER enzyme glucose 6-phosphatase. Insulin-sensitive tissues such as skeletal muscle and adipose tissue contain GLUT-4. When the concentration of blood glucose increases in response to food intake, pancreatic GLUT-2 molecules mediate an increase in glucose uptake which leads to increased insulin secretion. Recent evidence has shown that the cell surface receptor for the human T cell leukemia virus (HTLV) is the ubiquitous GLUT-1.

Hepatocytes, unlike most other cells, are freely permeable to glucose and are, therefore, essentially unaffected by the action of insulin at the level of increased glucose uptake. When blood glucose levels are low the liver does not compete with other tissues for glucose since the extrahepatic uptake of glucose is stimulated in response to insulin. Conversely, when blood glucose levels are high extrahepatic needs are satisfied and the liver takes up glucose for conversion into glycogen for future needs. Under conditions of high blood glucose, liver glucose levels will be high and the activity of glucokinase will be elevated. The G6P produced by glucokinase is rapidly converted to G1P by phosphoglucomutase, where it can then be incorporated into glycogen.
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The World Health Organization recognizes three main forms of diabetes: type 1, type 2, and gestational diabetes (occurring during pregnancy),[ which have similar signs, symptoms, and consequences, but different causes and population distributions. Ultimately, all forms are due to the beta cells of the pancreas being unable to produce sufficient insulin to prevent hyperglycemia Type 1 is usually due to autoimmune destruction of the pancreatic beta cells which produce insulin. Type 2 is characterized by tissue-wide insulin resistance and varies widely; it sometimes progresses to loss of beta cell function. Gestational diabetes is similar to type 2 diabetes, in that it involves insulin resistance; the hormones of pregnancy cause insulin resistance in those women genetically predisposed to developing this condition.

Types 1 and 2 are incurable chronic conditions, but have been treatable since insulin became medically available in 1921, and are nowadays usually managed with a combination of dietary treatment, tablets (in type 2) and, frequently, insulin supplementation. Gestational diabetes typically resolves with delivery.

Diabetes can cause many complications. Acute complications (hypoglycemia, ketoacidosis or nonketotic hyperosmolar coma) may occur if the disease is not adequately controlled. Serious long-term complications include cardiovascular disease (doubled risk), chronic renal failure (diabetic nephropathy is the main cause of dialysis in developed world adults), retinal damage (which can lead to blindness and is the most significant cause of adult blindness in the non-elderly in the developed world), nerve damage (of several kinds), and microvascular damage, which may cause erectile dysfunction (impotence) and poor healing. Poor healing of wounds, particularly of the feet, can lead to gangrene which can require amputation — the leading cause of non-traumatic amputation in adults in the developed world. Adequate treatment of diabetes, as well as increased emphasis on blood pressure control and lifestyle factors (such as smoking and keeping a healthy body weight), may improve the risk profile of most aforementioned complications.

 

Digestion & Absorption of Proteins & Carbohydrates

Digestion and Absorption of Proteins

General Information:

1. Humans must ingest proteins, carbohydrates and lipids to maintain tissue and organ functions.

2. Most of these nutrients consist of large polymers that must be broken down before they can be made available to the intestinal cells for transport.

3. Dietary proteins are cleaved by hydrolases with specificity for the peptide bond (peptidases).

4. Endopeptidases (aka Proteases): attack internal protein bonds liberating large peptide fragments.

Exopeptidases: cleave off one amino acid at a time from the….

NH3+, aminopeptidases or COO terminus, carboxypeptidase.

5. Endo- and Exopeptidases work in concert

The Big Picture:

Protein Digestion and Absorption

http://www.gravitywaves.com/chemistry/CHE452/images/ProtDig.gif

Gastric (Stomach) Digestion:

1. Gastric HCl is responsible for the low pH <2 of gastric juice.

2. Gastric acid kills microorganisms and denatures dietary proteins preparing them for hydrolysis by proteases.

3. Gastric juices contain the acid stable proteases of the pepsin family, which produce large peptide fragments and some free amino acids.

4. Protein digestion at this stage is partial, as the amino acids enter the duodenum, they trigger the release of cholectystokinin-pancreozymin (CCK-PZ) into the bloodstream.

This release initiates the secretion of protease zymogens from the pancreas and releases of enteropeptidase in the gut.

Pancreatic Proteases:

1. The pancreatic juice is rich in the proenzymes of endopeptidase and carboxypeptidases.

2. Enteropeptidase activates pancreatic trypsinogen to trypsin.

3. Trypsin autocatalytically activates more trypsinogen and other proenzymes, liberating chymotrypsin, elastase and the carboxypeptidases A and B.

Secretion and Activation of Pancreatic Proteases:

http://www.gravitywaves.com/chemistry/CHE452/images/CHODig.gif

Digestion at the Brush Border (surface of intestinal epithelial cells):

1. Since pancreatic juice does not contain appreciable aminopeptidase activity, final digestion of di- and small peptides depends on brush border enzymes.

2. The surface of intestinal epithelial cells is rich in endopeptidases and aminopeptidases.

3. The end products of cell surface digestion are free amino acids and di- and tripeptides.

Absorption:

1. Following digestion, amino acids and small peptides are co-absorbed w/ sodium via group specific amino acid or peptide transport systems.

2. These processes are carrier mediated, discriminating betweeatural, L amino acids and D-amino acids, require energy (from the Na+ gradient, Na-K ATPase) and physiologic temperatures.

At least five brush border transport systems exist:

1. neutral amino acids (uncharged aliphatic and aromatic)

2. basic amino acids (Lys, Arg, Cys, Cys-Cys)

3. acidic amino acids (Asp, Glu)

4. imino acids (Pro), Hydroxyproline)

5. di- and tripeptides

Clinical Correlates:

1. Hartnup Disease:

Genetic defect in the neutral amino acid transporter.

Symptoms: dermatitis due to tryptophan malabsorption (“niacin” flush)

Consequences: not serious di- and tripeptide absorption supply minimal amounts of dietarily essential neutral amino acids.

2. Cystinuria:

Precursor to kidney stones

Symptoms: painful kidney stone formation due to malabsoprtion of cystine (two disulfide linked cysteines)

3. Sprue:

Destruction and flattening of the intestinal villi resulting in generalized malabsorption.

Causes: bacterial infection or gluten (contained in certain grains such as wheat and barley) sensitivity.

Digestion and Absorption of Carbohydrates

General Information:

1. Carbohydrates provide a major component of the daily caloric requirement, ~40%.

2.Distinguish between mono-, di- and polysaccharides.

Monosaccharides- do not need hydrolysis prior to absorption.

Disaccharides- require brush border enzymes.

Polysaccharides- require brush border enzymes, as well as, pancreatic amylase and salivary amylase for digestion.

Starch:

Hydrolyzed by -amylase into Maltotriose, -Limit Dextrin, Maltose, Glucose

-1,4-glucosidic linkages (non-branching, amylose) and branched chains -1,6 linkages (branch points, amylopectin)

-amylase:

Present in saliva and pancreatic juice.

Specific for internal -1,4-glucosidic bonds.

Brush Border Carbohydrate Digestion:

Final hydrolysis of di- and oligosaccharides to monosaccharides is carried out by -glucosidases on the surface of the small intestine.

http://www.gravitywaves.com/chemistry/CHE452/images/CHOEnzTable.gif

Monosaccharides are absorbed by carrier mediated transport.

At least two types are known:

1. Na+monosaccharide transporter

2. Na+ independent, diffusion type monosaccharide transport system

http://www.gravitywaves.com/chemistry/CHE452/images/LacSucDig.gif

Undigested Carbohydrates:

1. Di-, oligo- and polysaccharides that are not hydrolyzed by -amylase and/or brush border enzymes cannot be absorbed.

2. These carbohydrates reach the lower tract of the intestine which contains bacteria.

3. The bacteria utilize many of the remaining carbohydrate, metabolizing them and producing by- products such as: hydrogen gas, methane and carbon dioxide.

 

Digestion of Carbohydrates

Carbohydrates

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Carbohydrates are one of three macronutrients that provide the body with energy (protein and fats being the other two). The chemical compounds in carbohydrates are found in both simple and complex forms, and in order for the body to use carbohydrates for energy, food must undergo digestion, absorption, and glycolysis. It is recommended that 55 to 60 percent of caloric intake come from carbohydrates.

Chemical Structure

Carbohydrates are a main source of energy for the body and are made of carbon, hydrogen, and oxygen. Chlorophyll in plants absorbs light energy from the sun. This energy is used in the process of photosynthesis, which allows green plants to take in carbon dioxide and release oxygen and allows for the production of carbohydrates. This process converts the sun’s light energy into a form of chemical energy useful to humans. Plants transform carbon dioxide (CO2) from the air, water (H2O) from the ground, and energy from the sun into oxygen (O2) and carbohydrates (C6H12O6) (6 CO2 + 6 H2O + energy = C6H12O6 + 6 O2). Most carbohydrates have a ratio of 1:2:1 of carbon, hydrogen, and oxygen, respectively.

Humans and other animals obtain carbohydrates by eating foods that contain them. In order to use the energy contained in the carbohydrates, humans must metabolize, or break down, the structure of the molecule in a process that is opposite that of photosynthesis. It starts with the carbohydrate and oxygen and produces carbon dioxide, water, and energy. The body utilizes the energy and water and rids itself of the carbon dioxide.

Simple Carbohydrates

Simple carbohydrates, or simple sugars, are composed of monosaccharide or disaccharide units. Common monosaccharides (carbohydrates composed of single sugar units) include glucose, fructose, and galactose. Glucose is the most common type of sugar and the primary form of sugar that is stored in the body for energy. It sometimes is referred to as blood sugar or dextrose and is of particular importance to individuals who have diabetes or hypoglycemia. Fructose, the primary sugar found in fruits, also is found in honey and high-fructose corn syrup (in soft drinks) and is a major source of sugar in the diet of Americans. Galactose is less likely than glucose or fructose to be found iature. Instead, it often combines with glucose to form the disaccharide lactose, often referred to as milk sugar. Both fructose and galactose are metabolized to glucose for use by the body.

Oligosaccharides are carbohydrates made of two to ten monosaccharides. Those composed of two sugars are specifically referred to as disaccharides, or double sugars. They contain two monosaccharides bound by either an alpha bond or a beta bond. Alpha bonds are digestible by the human body, whereas beta bonds are more difficult for the body to break down.

There are three particularly important disaccharides: sucrose, maltose, and lactose. Sucrose is formed when glucose and fructose are held together by an alpha bond. It is found in sugar cane or sugar beets and is refined to make granulated table sugar. Varying the degree of purification alters the

SUGAR COMPARISON

SUGAR COMPARISON

Sugar

Carbohydrate

Monosaccharide or disaccharide

Additional information

Beet sugar (cane sugar)

Sucrose

Disaccharide (fructose and glucose)

Similar to white and powdered sugar, but varied degree of purification

Brown sugar

Sucrose

Disaccharide (fructose and glucose)

Similar to white and powdered sugar, but varied degree of purification

Corn syrup

Glucose

Monosaccharide

 

Fruit sugar

Fructose

Monosaccharide

Very sweet

High-fructose corn syrup

Fructose

Monosaccharide

Very sweet and inexpensive
Added to soft drinks and canned or frozen fruits

Honey

Fructose and glucose

Monosaccharides

 

Malt sugar

Maltose

Disaccharide (glucose and glucose)

Formed by the hydrolysis of starch, but sweeter than starch

Maple syrup

Sucrose

Disaccharide (fructose and glucose)

 

Milk sugar

Lactose

Disaccharide (glucose and galactose)

Made in mammary glands of most lactating animals

Powdered sugar

Sucrose

Disaccharide (fructose and glucose)

Similar to white and brown sugar, but varied degree of purification

White sugar

Sucrose

Disaccharide (fructose and glucose)

Similar to brown and powdered sugar, but varied degree of purification

SOURCE: Mahan and Escott-Stump, 2000; Northwestern University; Sizer and Whitney, 1997; and Wardlaw and Kessel, 2002.

final product, but white, brown, and powdered sugars all are forms of sucrose. Maltose, or malt sugar, is composed of two glucose units linked by an alpha bond. It is produced from the chemical decomposition of starch, which occurs during the germination of seeds and the production of alcohol. Lactose is a combination of glucose and galactose. Because it contains a beta bond, it is hard for some individuals to digest in large quantities. Effective digestion requires sufficient amounts of the enzyme lactase.

Complex Carbohydrates

Complex carbohydrates, or polysaccharides, are composed of simple sugar units in long chains called polymers. Three polysaccharides are of particular importance in human nutrition: starch, glycogen, and dietary fiber.

Starch and glycogen are digestible forms of complex carbohydrates made of strands of glucose units linked by alpha bonds. Starch, often contained in seeds, is the form in which plants store energy, and there are two types: amylose and amylopectin. Starch represents the main type of digestible complex carbohydrate. Humans use an enzyme to break down the bonds linking glucose units, thereby releasing the sugar to be absorbed into the bloodstream. At that point, the body can distribute glucose to areas that need energy, or it can store the glucose in the form of glycogen.

Glycogen is the polysaccharide used to store energy in animals, including humans. Like starch, glycogen is made up of chains of glucose linked by alpha bonds; but glycogen chains are more highly branched than starch. It is this highly branched structure that allows the bonds to be more quickly broken down by enzymes in the body. The primary storage sites for glycogen in the human body are the liver and the muscles.

Another type of complex carbohydrate is dietary fiber. In general, dietary fiber is considered to be polysaccharides that have not been digested at the point of entry into the large intestine. Fiber contains sugars linked by bonds that cannot be broken down by human enzymes, and are therefore

Pastas and whole-grain breads contain complex carbohydrates, which are long strands of glucose molecules. Nutritionists recommend that 55&#x2013;60 percent of calories come from carbohydrates, and especially complex carbohydrates. [Photograph by James Noble. Corbis. Reproduced by permission.]

Pastas and whole-grain breads contain complex carbohydrates, which are long strands of glucose molecules. Nutritionists recommend that 55–60 percent of calories come from carbohydrates, and especially complex carbohydrates.

[Photograph by James Noble. Corbis. Reproduced by permission.]

labeled as indigestible. Because of this, most fibers do not provide energy for the body. Fiber is derived from plant sources and contains polysaccharides such as cellulose, hemicellulose, pectin, gums, mucilages, and lignins.

The indigestible fibers cellulose, hemicellulose, and lignin make up the structural part of plants and are classified as insoluble fiber because they usually do not dissolve in water. Cellulose is a nonstarch carbohydrate polymer made of a straight chain of glucose molecules linked by beta bonds and can be found in whole-wheat flour, bran, and vegetables. Hemicellulose is a nonstarch carbohydrate polymer made of glucose, galactose, xylose, and other monosaccharides; it can be found in bran and whole grains. Lignin, a noncarbohydrate polymer containing alcohols and acids, is a woody fiber found in wheat bran and the seeds of fruits and vegetables.

In contrast, pectins, mucilages, and gums are classified as soluble fibers because they dissolve or swell in water. They are not broken down by human enzymes, but instead can be metabolized (or fermented) by bacteria present in the large intestine. Pectin is a fiber made of galacturonic acid and other monosaccharides. Because it absorbs water and forms a gel, it is often used in jams and jellies. Sources of pectin include citrus fruits, apples, strawberries, and carrots. Mucilages and gums are similar in structure. Mucilages are dietary fibers that contain galactose, manose, and other monosaccharides; and gums are dietary fibers that contain galactose, glucuronic acid, and other monosaccharides. Sources of gums include oats, legumes, guar, and barley.

Digestion and Absorption

Carbohydrates must be digested and absorbed in order to transform them into energy that can be used by the body. Food preparation often aids in the digestion process. When starches are heated, they swell and become easier for the body to break down. In the mouth, the enzyme amylase, which is contained in saliva, mixes with food products and breaks some starches into smaller units. However, once the carbohydrates reach the acidic environment of the stomach, the amylase is inactivated. After the carbohydrates have passed through the stomach and into the small intestine, key digestive enzymes are secreted from the pancreas and the small intestine where most digestion and absorption occurs. Pancreatic amylase breaks starch into disaccharides and small polysaccharides, and enzymes from the cells of the small-intestinal wall break any remaining disaccharides into their monosaccharide components. Dietary fiber is not digested by the small intestine; instead, it passes to the colon unchanged.

Sugars such as galactose, glucose, and fructose that are found naturally in foods or are produced by the breakdown of polysaccharides enter into absorptive intestinal cells. After absorption, they are transported to the liver where galactose and fructose are converted to glucose and released into the bloodstream. The glucose may be sent directly to organs that need energy, it may be transformed into glycogen (in a process called glycogenesis) for storage in the liver or muscles, or it may be converted to and stored as fat.

Glycolysis

http://www.youtube.com/watch?v=O5eMW4b29rg&feature=related

 

The molecular bonds in food products do not yield high amounts of energy when broken down. Therefore, the energy contained in food is released within cells and stored in the form of adenosine triphosphate (ATP), a high-energy compound created by cellular energy-production systems. Carbohydrates are metabolized and used to produce ATP molecules through a process called glycolysis.

Glycolysis breaks down glucose or glycogen into pyruvic acid through enzymatic reactions within the cytoplasm of the cells. The process results in the formation of three molecules of ATP (two, if the starting product was glucose). Without the presence of oxygen, pyruvic acid is changed to lactic acid, and the energy-production process ends. However, in the presence of oxygen, larger amounts of ATP can be produced. In that situation, pyruvic acid is transformed into a chemical compound called acetyle coenzyme A, a compound that begins a complex series of reactions in the Krebs Cycle and the electron transport system. The end result is a net gain of up to thirty-nine molecules of ATP from one molecule of glycogen (thirty-eight molecules of ATP if glucose was used). Thus, through certain systems, glucose can be used very efficiently in the production of energy for the body.

Recommended Intake

At times, carbohydrates have been incorrectly labeled as “fattening.” Evidence actually supports the consumption of more, rather than less, starchy foods. Carbohydrates have four calories per gram, while dietary fats contribute nine per gram, so diets high in complex carbohydrates are likely to provide fewer calories than diets high in fat. Recommendations are for 55 to 60 percent of total calories to come from carbohydrates (approximately 275 to 300 grams for a 2,000-calorie diet). The majority of carbohydrate calories should come from complex rather than simple carbohydrates. Of total caloric intake, approximately 45 to 50 percent of calories should be from complex carbohydrates, and 10 percent or less from simple carbohydrates.

Low-Carb Diets

Low-carbohydrate diets, such as the Atkins and South Beach diets, are based on the proposition that it’s not fat that makes you fat. Allowing dieters to eat steak, butter, eggs, bacon, and other high-fat foods, these diets instead outlaw starches and refined carbohydrates on the theory that they are metabolized so quickly that they lead to hunger and overeating. This theory, which was first popularized in the nineteenth century, came under scathing criticism from the medical establishment during the early 1970s when Dr. Robert Atkins published the phenomenally popular low-carb diet bearing his name. According to the American Medical Association (AMA), the Atkins diet was a “bizarre regimen” that advocated “an unlimited intake of saturated fats and cholesterol-rich foods” and therefore presented a considerable risk of heart disease. Most doctors recommended instead a diet low in fat and high in carbohydrates, with plenty of grains, fruits, and vegetables and limited red meat or dairy products. This became the received wisdom during the 1980s, at the same time that the U.S. waistline began to expand precipitously. As dieters found that weight loss was difficult to maintain on a low-fat diet, low-carb diets regained popularity—with as many as 30 million people trying a low-carb diet in 2003. Several small-scale studies began to suggest that a low-carb diet may indeed be effective and may not have the deleterious effects its detractors have claimed; other research found that any benefits of a low-carb diet are short-lived, and that the negative effects will take decades to become evident. The National Institutes of Health has pledged $2.5 million for a five-year study of the Atkins diet with 360 subjects. While the results of this and other large-scale studies are awaited, many researchers stress that the key issue in maintaining a healthy weight is the number of calories consumed, not the type of calories. The National Academy of Sciences recommends that adults obtain 45 to 65 percent of their calories from carbohydrates, 20 to 35 percent from fat, and 10 to 35 percent from protein.

It is important to consume a minimum amount of carbohydrates to prevent ketosis, a condition resulting from the breakdown of fat for energy in the absence of carbohydrates. In this situation, products of fat breakdown, called ketone bodies, build up in the blood and alter normal pH balance. This can be particularly harmful to a fetus. To avoid ketosis, daily carbohydrate intake should include a minimum of 50 to 100 grams. In terms of dietary fiber, a minimum intake of 20 to 35 grams per day is recommended.

Exchange System

The exchange system is composed of lists that describe carbohydrate, fat, and protein content, as well as caloric content, for designated portions of specific foods. This system takes into account the presence of more than one type of nutrient in any given food. Exchange lists are especially useful for individuals who require careful diet planning, such as those who monitor intake of calories or certaiutrients. It is particularly useful for diabetics, for whom carbohydrate intake must be carefully controlled, and was originally developed for planning diabetic diets.

Diabetes, Carbohydrate-Modified Diets, and Carbohydrate Counting

Diabetes is a condition that alters the way the body handles carbohydrates. In terms of diet modifications, diabetics can control blood sugar levels by appropriately managing the carbohydrates, proteins, and fats in their meals. The amount of carbohydrates, not necessarily the source, is the primary issue. Blood glucose levels after a meal can be related to the process of food preparation, the amount of food eaten, fat intake, sugar absorption, and the combination of foods in the meal or snack.

One method of monitoring carbohydrate levels—carbohydrate counting—assigns a certaiumber of carbohydrate grams or exchanges to specific foods. Calculations are used to determine insulin need, resulting in better control of blood glucose levels with a larger variety of foods. Overall, diabetic diets can include moderate amounts of sugar, as long as they are carefully monitored.

Glycolysis (detailed)

http://www2.carthage.edu/~pfaffle/170/Glycolysis.jpg

Nomenclature of carbohydrates – the naming of carbohydrates – much more information than most of us need

1. Glucose – a monosaccharide – what you measure in your blood.

The chemical formula for glucose is C6H12O6.

http://www.peoriaendocrine.com/images/diabetes_lecture/glucose.GIF

2. Sucrose – a disaccharide – table sugar. Is made of one molecule of glucose and one fructose.

http://www.peoriaendocrine.com/images/diabetes_lecture/sucrose.GIF

3. Starch – amylose and amylopectin – a polysaccharide – potatoes.

Amylose is a straight chain of glucose molecules linked together.

Amylopectin is a branched chain of glucose molecules.

 

http://www.peoriaendocrine.com/images/diabetes_lecture/starch.JPG

 

4. Glycolysis is the breakdown of glucose to release its energy in the form of ATP.

Carbohydrate metabolism – a slide presentation from the University of Akron – a little

more chemistry than most people will want.

Pentose Phosphate Pathway

The Pentose Phosphate Pathway (also called Phosphogluconate Pathway, or Hexose Monophosphate Shunt) is depicted with structures of intermediates in Fig. 23-25 p. 863 of Biochemistry, by Voet & Voet, 3rd Edition. The linear portion of the pathway carries out oxidation and decarboxylation of glucose-6-phosphate, producing the 5-C sugar ribulose-5-phosphate.

Glucose-6-phosphate Dehydrogenase catalyzes oxidation of the aldehyde (hemiacetal), at C1 of glucose-6-phosphate, to a carboxylic acid in ester linkage (lactone). NADP+ serves as electron acceptor.

6-Phosphogluconolactonase catalyzes hydrolysis of the ester linkage (lactone) resulting in ring opening. The product is 6-phosphogluconate. Although ring opening occurs in the absence of a catalyst, 6-Phosphogluconolactonase speeds up the reaction, decreasing the lifetime of the highly reactive, and thus potentially toxic, 6-phosphogluconolactone.

Phosphogluconate Dehydrogenase catalyzes oxidative decarboxylation of 6-phosphogluconate, to yield the 5-C ketose ribulose-5-phosphate. The hydroxyl at C3 (C2 of the product) is oxidized to a ketone. This promotes loss of the carboxyl at C1 as CO2NADP+ again serves as oxidant (electron acceptor). 

 

Reduction of NADP+ (as with NAD+) involves transfer of 2e plus 1H+ to the nicotinamide moiety.

 

NAD+ and NADP+ differ only in the presence of an extra phosphate on the adenosine ribose of NADP+. This difference has little to do with redox activity, but is recognized by substrate-binding sites of enzymes. It is a mechanism for separation of catabolic and synthetic pathways.

NADPH, a product of the Pentose Phosphate Pathway, functions as a reductant in various synthetic (anabolic) pathways, including fatty acid synthesis. 

NAD+ serves as electron acceptor in catabolic pathways in which metabolites are oxidized. The resultant NADH is reoxidized by the respiratory chain, producing ATP.

Regulation:
Glucose-6-phosphate Dehydrogenase
is the committed step of the Pentose Phosphate Pathway. This enzyme is regulated by availability of the substrate NADP+. As NADPH is utilized in reductive synthetic pathways, the increasing concentration of NADP+ stimulates the Pentose Phosphate Pathway, to replenish NADPH. 

The remainder of the Pentose Phosphate Pathway accomplishes conversion of the 5-C ribulose-5-phosphate to the 5-C product ribose-5-phosphate, or to the 3-C glyceraldehyde-3-phosphate and the 6-C fructose-6-phosphate (reactions 4 to 8 p. 863). 

Additional enzymes include Ribulose-5-phosphate Epimerase, Ribulose-5-phosphate Isomerase, Transketolase, and Transaldolase.

·     Epimerase interconverts the stereoisomers ribulose-5-phosphate and xylulose-5-phosphate. 

·     Isomerase converts the ketose ribulose-5-phosphate to the aldose ribose-5-phosphate. 

·     Both reactions involve deprotonation to form an endiolate intermediate, followed by specific reprotonation to yield the product. Both reactions are reversible.

Transketolase and Transaldolase catalyze transfer of 2-C and 3-C molecular fragments respectively, in each case from a ketose donor to an aldose acceptor. D. E. Nicholson has suggested that the names of these enzymes should be changed, since Transketolase actually transfers an aldol moiety (glycoaldehyde) and Transaldolase actually transfers a ketol moiety (dihydroxyacetone). However the traditional enzyme names are used here.

Transketolase transfers a 2-C fragment from xylulose-5-phosphate to either ribose-5-phosphate or erythrose-4-phosphate.

 

 

Thiamine pyrophosphate binds at the active sites of enzymes in a “V” conformation. The amino group of the aminopyrimidine moiety is close to the dissociable proton, and serves as the proton acceptor. This proton transfer is promoted by a glutamate residue adjacent to the pyrimidine ring.

The thiazolium carbanion (ylid) that results from proton dissociation reacts with the carbonyl C of xylulose-5-P to form an addition compound. 

The positively charged N in the thiazole ring acts as an electron sink, promoting C-C bond cleavage. The 3-C aldose glyceraldehyde-3-phosphate is released. A 2-C fragment remains on TPP.

Completion of the reaction is by reversal of these steps. The 2-C fragment condenses with one of the aldoses erythrose-4-phosphate (4-C) or ribose-5-phosphate (5-C) to form a 6-C or 7-C ketose-phosphate product.

Transfer of the 2-C fragment to the 5-C aldose ribose-5-phosphate yields sedoheptulose-7-phosphate (see above). Transfer instead to the 4-C aldose erythrose-4-phosphate yields fructose-6-phosphate. See also diagram p. 865. 

 

Explore at right the structure of  Transketolase with bound substrate erythrose-4-phosphate.



 

Transaldolase catalyzes transfer of a 3-C dihydroxyacetone moiety, from sedoheptulose-7-phosphate to glyceraldehyde-3-phosphate.

 

The e-amino group of an active site lysine residue reacts with the carbonyl C of sedoheptulose-7-phosphate to form a protonated Schiff base intermediate.

Aldol cleavage results in release of erythrose-4-phosphate. The Schiff base stabilizes the carbanion on C3.

Completion of the reaction occurs by reversal, as the carbanion attacks instead the aldehyde carbon of the 3-carbon aldose glyceraldehyde-3-phosphate to yield the 6-carbon fructose-6-phosphate. See also diagram p. 866. 

 

Explore at right the structure of E. coli Transaldolase, crystallized with a modified active site intermediate. The structure of human Transaldolase has also been determined, and it exhibits a similar a,b barrel structure.



 

The flow of intermediates containing 15 C atoms through Pentose Phosphate Pathway reactions by which 5-C sugars are converted to 3-C and 6-C sugars is summarized in the diagram at right and balance sheet below.

C5 + C5 C3 + C7 (Transketolase)

C3 + C7 C6  + C4  (Transaldolase)

C5 + C4 C6 + C3 (Transketolase)

_______________________________________

3 C5 2C6 + C3   (Overall)

 

Glucose-6-phosphate may be regenerated from either the 3-C product glyceraldehyde-3-phosphate or the 6-C product fructose-6-phosphate, via enzymes of Gluconeogenesis.
 

In the diagram at right, IS = Isomerase, EP = Epimerase, TK = Transketolase, TA = Transaldolase.

Depending on relative needs of a cell for ribose-5-phosphate, NADPH, and ATP, the Pentose Phosphate Pathway can operate in various modes, to maximize different products. There are three major scenarios:

1. Ribulose-5-phosphate may be converted to ribose-5-phosphate, a substrate for synthesis of nucleotides and nucleic acids. The pathway also produces some NADPH.

2. Glyceraldehyde-3-phosphate and fructose-6-phosphate, formed from the 5-carbon sugar phosphates, may be converted to glucose-6-phosphate for reentry into the linear portion of the Pentose Phosphate Pathway, maximizing formation of NADPH.

3. Glyceraldehyde-3-phosphate and fructose-6-phosphate, formed from the 5-carbon sugar phosphates, may enter Glycolysis, for synthesis of ATP. The pathway also produces some NADPH.

Ribose-1-phosphate generated during catabolism of nucleosides also enters the Glycolytic pathway in this way, after first being converted to ribose-5-phosphate. Thus the Pentose Phosphate Pathway serves as an entry into Glycolysis for both 5-carbon and 6-carbon sugars.

 

Glutathione is a tripeptide that includes a glutamate residue linked by an isopeptide bond involving the side-chain carbonyl group. Its functional group is a cysteine thiol.

One role of glutathione is degradation of hydroperoxides that arise spontaneously in the oxygen-rich environment within red blood cells. Hydroperoxides can react with double bonds in fatty acid moieties of membrane lipids, making membranes leaky.

Glutathione Peroxidase catalyzes degradation of organic hydroperoxides by reduction, as two glutathione molecules are oxidized to a disulfide. In the reaction summary below, glutathione is represented as GSH.

2 GSH + ROOH GSSG + ROH + H2O

Glutathione Peroxidase contains the trace element selenium as a functional group. The enzyme’s primary structure includes an analog of cysteine, selenocysteine, with Se replacing S.

Regeneration of  reduced glutathione requires NADPH, produced within erythrocytes in the Pentose Phosphate Pathway. Glutathione Reductase catalyzes:

GSSG + NADPH + H+ 2 GSH + NADP+

Genetic deficiency of Glucose-6-phosphate Dehydrogenase can lead to a hemolytic anemia, because of an inadequate supply of NADPH within red blood cells. The effect of a partial deficiency of Glucose-6-phosphate Dehydrogenase activity is exacerbated by substances that lead to increased production of peroxides (e.g., the antimalarial primaquine).

 

 

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