carbohydrate

/kahr'boh huy"drayt, -beuh-/, n.

any of a class of organic compounds that are polyhydroxy aldehydes or polyhydroxy ketones, or change to such substances on simple chemical transformations, as hydrolysis, oxidation, or reduction, and that form the supporting tissues of plants and are important food for animals and people.

[1865-70; CARBO- + HYDRATE]

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Any member of a very abundant and widespread class of natural organic compounds that includes sugars, starch, and cellulose.

They are commonly classified as monosaccharides (simple sugars; e.g., glucose, fructose), disaccharides (2-unit sugars; e.g., sucrose, lactose), oligosaccharides (3–10 or so sugars), and polysaccharides (large molecules with up to 10,000 monosaccharide units, including cellulose, starch, and glycogen). Green plants produce carbohydrates by photosynthesis. In most animals, carbohydrates are the quickly accessible reservoir of energy, and oxidation (see oxidation-reduction) of glucose in tissues supplies energy for metabolism. Many (but by no means all) carbohydrates have the general chemical formula C_n(H₂O)_n. The carbon (C) atoms are bonded to hydrogen atoms (―H), hydroxyl groups (―OH; see functional group), and carbonyl groups (―C=O), whose combinations, order, and geometric arrangement lead to a large number of isomers with the same chemical formula but different properties. The class is further enlarged because each isomer has various derivatives: uronic acids, sugars with an oxidized group; sugar alcohols, sugars with a reduced group; glycosides, compounds of sugars with other molecules containing a hydroxyl group; and amino sugars, sugars with an amino group (see amino acid).

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▪ biochemistry

Introduction

 class of naturally occurring compounds and derivatives formed from them. In the early part of the 19th century, substances such as wood, starch, and linen were found to be composed mainly of molecules containing atoms of carbon (C), hydrogen (H), and oxygen (O), and to have the general formula C₆H₁₂O₆; other organic molecules with similar formulas were found to have a similar ratio of hydrogen to oxygen. The general formula C_x(H₂O)_x is commonly used to represent many carbohydrates, which means “watered carbon.”

      Carbohydrates are probably the most abundant and widespread organic substances in nature, and they are essential constituents of all living things. Carbohydrates are formed by green plants from carbon dioxide and water during the process of photosynthesis. Carbohydrates serve organisms as energy sources and as essential structural components; in addition, part of the structure of nucleic acids, which contain genetic information, consists of carbohydrate.

General features

Classification and nomenclature

      Although a number of classification schemes have been devised for carbohydrates, the division into four major groups—monosaccharides, disaccharides, oligosaccharides, and polysaccharides—used here is among the most common. Most monosaccharides (monosaccharide), or simple sugars (sugar), are found in grapes, other fruits, and honey. Although they can contain from three to nine carbon atoms, the most common representatives consist of five or six joined together to form a chainlike molecule. Three of the most important simple sugars, glucose—also known as dextrose, grape sugar, and corn sugar—fructose (fruit sugar (fructose)), and galactose, have the same molecular formula, (C₆H₁₂O₆), but, because their atoms have different structural arrangements, the sugars have different characteristics; i.e., they are isomers. Slight changes in structural arrangements are detectable by living things and influence the biological significance of isomeric compounds. It is known, for example, that the degree of sweetness of various sugars differs according to the arrangement of the hydroxyl groups (−OH) that compose part of the molecular structure; a direct correlation that may exist between taste and any specific structural arrangement, however, has not yet been established—that is, it is not yet possible to predict the taste of a sugar by knowing its specific structural arrangement. The energy in the chemical bonds of glucose indirectly supplies most living things with a major part of the energy that is necessary for them to carry on their activities. Galactose, which is rarely found as a simple sugar, is usually combined with other simple sugars in order to form larger molecules.

      Two molecules of a simple sugar that are linked to each other form a disaccharide, or double sugar. The disaccharide sucrose, or table sugar, consists of one molecule of glucose and one molecule of fructose; the most familiar sources of sucrose are sugar beets and cane sugar. Milk sugar, or lactose, and maltose are also disaccharides. Before the energy in disaccharides can be utilized by living things, the molecules must be broken down into their respective monosaccharides.

      Oligosaccharides (oligosaccharide), which consist of three to six monosaccharide units, are rather infrequently found in natural sources, although a few plant derivatives have been identified.

      Polysaccharides (polysaccharide) (the term means many sugars) represent most of the structural and energy-reserve carbohydrates found in nature. Large molecules that may consist of as many as 10,000 monosaccharide units linked together, polysaccharides vary considerably in size, in structural complexity, and in sugar content; several hundred distinct types have thus far been identified. cellulose, the principal structural component of plants, is a complex polysaccharide comprising many glucose units linked together; it is the most common polysaccharide. The starch found in plants and the glycogen found in animals also are complex glucose polysaccharides. Starch (from the old English word stercan meaning “to stiffen”) is found mostly in seeds, roots, and stems, where it is stored as an available energy source for plants. Plant starch may be processed into such foods as bread, or it may be consumed directly—as potatoes, for instance. Glycogen, which consists of branching chains of glucose molecules, is formed in the liver and muscles of higher animals and is stored as an energy source.

      The generic nomenclature ending for the monosaccharides is -ose; thus, the term pentose (pent = five) is used for monosaccharides containing five carbon atoms, and hexose (hex = six) is used for those containing six. In addition, because the monosaccharides contain a chemically reactive group that is either an aldehydo group

, they are frequently referred to as aldopentoses or ketopentoses or aldohexoses or ketohexoses; in the examples below, the aldehydo group is at position 1 of the aldopentose, the keto group is at position 2 of the ketohexose. Glucose is an aldohexose—i.e., it contains six carbon atoms, and the chemically reactive group is an aldehydo group.

Biological significance

      The importance of carbohydrates to living things can hardly be overemphasized. The energy stores of most animals and plants are both carbohydrate and lipid in nature; carbohydrates are generally available as an immediate energy source, whereas lipids act as a long-term energy resource and tend to be utilized at a slower rate. Glucose, the prevalent uncombined, or free, sugar circulating in the blood of higher animals, is essential to cell function. The proper regulation of glucose metabolism is of paramount importance to survival.

      The ability of ruminants, such as cattle, sheep, and goats, to convert the polysaccharides present in grass and similar feeds into protein provides a major source of protein for man. A number of medically important antibiotics, such as streptomycin, are carbohydrate derivatives. The cellulose in plants is used to manufacture paper, wood for construction, and fabrics.

Role in the biosphere

      The essential process in the biosphere, the portion of the Earth in which life can occur, that has permitted the evolution of life as it now exists is the conversion by green plants of carbon dioxide from the atmosphere into carbohydrates, using light energy from the Sun. This process, called photosynthesis, results in both the release of oxygen gas into the atmosphere and the transformation of light energy into the chemical energy of carbohydrates. The energy stored by plants during the formation of carbohydrates is used by animals to carry out mechanical work and to perform biosynthetic activities.

      All green plants apparently photosynthesize in the same way, yielding as an immediate product the compound 3-phosphoglyceric acid; the formula, in which P represents phosphorus, is illustrated below.

      This compound then is transformed into cell-wall components such as cellulose, varying amounts of sucrose, and starch—depending on the plant type—and a wide variety of polysaccharides, other than cellulose and starch, that function as essential structural components. For a detailed discussion of the process of photosynthesis, see photosynthesis.

Role in human nutrition

      The total caloric, or energy, requirement for an individual depends on age, occupation, and other factors but generally ranges between 2,000 and 4,000 calories per 24-hour period (one calorie, as this term is used in nutrition (nutrition, human), is the amount of heat necessary to raise the temperature of 1,000 grams of water from 15° to 16° C [59° to 61° F]; in other contexts this amount of heat is called the kilocalorie). Carbohydrate that can be used by man produces four calories per gram as opposed to nine calories per gram of fat and four per gram of protein. In areas of the world where nutrition is marginal, a high proportion (approximately one to two pounds) of an individual's daily energy requirement may be supplied by carbohydrate, with most of the remainder coming from a variety of fat sources.

      Although carbohydrates may compose as much as 80 percent of the total caloric intake in the human diet, for a given diet, the proportion of starch to total carbohydrate is quite variable, depending upon the prevailing customs. In the Far East and in areas of Africa, for example, where rice or tubers such as manioc provide a major food source, starch may account for as much as 80 percent of the total carbohydrate intake. In a typical Western diet, 33 to 50 percent of the caloric intake is in the form of carbohydrate. Approximately half (i.e., 17 to 25 percent) is represented by starch; another third by table sugar (sucrose) and milk sugar (lactose); and smaller percentages by monosaccharides such as glucose and fructose, which are common in fruits, honey, syrups, and certain vegetables such as artichokes, onions, and sugar beets. The small remainder consists of bulk, or indigestible carbohydrate, which comprises primarily the cellulosic outer covering of seeds and the stalks and leaves of vegetables. (See also nutrition.)

Role in energy storage

      Starches, the major plant-energy-reserve polysaccharides used by man, are stored in plants in the form of nearly spherical granules that vary in diameter from about three to 100 micrometres (about .0001 to .004 inch). Most plant starches consist of a mixture of two components, amylose and amylopectin (see diagrams). As the diagrams show, the glucose molecules composing amylose have a straight-chain, or linear, structure; amylopectin has a branched-chain structure and is a somewhat more compact molecule. Several thousand glucose units may be present in a single starch molecule (each small circle represents one glucose molecule).

      In addition to granules, many plants have large numbers of specialized cells (tissue), called parenchymatous cells, the principal function of which is the storage of starch; examples of plants with these cells include root vegetables and tubers. The starch content of plants varies considerably; the highest concentrations are found in seeds and in cereal grains, which contain up to 80 percent of their total carbohydrate as starch. The amylose and amylopectin components of starch occur in variable proportions; most plant species store approximately 25 percent of their starch as amylose and 75 percent as amylopectin. This proportion can be altered, however, by selective-breeding techniques, and some varieties of corn have been developed that produce up to 70 percent of their starch as amylose, which is more easily digested by man than is amylopectin.

      In addition to the starches, some plants (e.g., the Jerusalem artichoke and the leaves of certain grasses, particularly rye grass) form storage polysaccharides composed of fructose units rather than glucose. Although the fructose polysaccharides can be broken down and used to prepare syrups, they cannot be digested by higher animals.

      Starches are not formed by animals; instead, they form a closely related polysaccharide, glycogen. Virtually all vertebrate and invertebrate animal cells, as well as those of numerous fungi and protozoans, contain some glycogen; particularly high concentrations of this substance are found in the liver and muscle cells of higher animals. The overall structure of glycogen, which is a highly branched molecule consisting of glucose units, has a superficial resemblance to that of the amylopectin component of starch, although the structural details of glycogen are significantly different. Under conditions of stress or muscular activity in animals, glycogen is rapidly broken down to glucose, which is subsequently used as an energy source. In this manner, glycogen acts as an immediate carbohydrate reserve. Furthermore, the amount of glycogen present at any given time, especially in the liver, directly reflects an animal's nutritional state; i.e., when adequate food supplies are available, both glycogen and fat reserves of the body increase, but when food supplies decrease or when the food intake falls below the minimum energy requirements, the glycogen reserves are depleted quite rapidly, while those of fat are used at a slower rate.

Role in plant and animal structure

      Whereas starches and glycogen represent the major reserve polysaccharides of living things, most of the carbohydrate found in nature occurs as structural components in the cell walls of plants. Carbohydrates in plant cell walls generally consist of several distinct layers, one of which contains a higher concentration of cellulose than the others. The physical and chemical properties of cellulose are strikingly different from those of the amylose component of starch.

      In most plants, the cell wall is about 0.5 micrometres thick and contains a mixture of cellulose, pentose-containing polysaccharides (pentosans), and an inert (chemically unreactive) plastic-like material called lignin. The amounts of cellulose and pentosan may vary; most plants contain between 40 and 60 percent cellulose, although higher amounts are present in the cotton fibre.

      Polysaccharides also function as major structural components in animals. Chitin, which is similar to cellulose, is found in insects and other arthropods. Other complex polysaccharides predominate in the structural tissues of higher animals.

Structural arrangements and properties

Stereoisomerism

      Studies by the German chemist Emil Fischer (Fischer, Emil) in the late 19th century showed that carbohydrates, such as fructose and glucose, with the same molecular (molecule) formulas but with different structural arrangements and properties (i.e., isomers) can be formed by relatively simple variations of their spatial, or geometric, arrangements. This type of isomerism, which is called stereoisomerism, exists in all biological systems; and, among carbohydrates, the simplest example is provided by the three-carbon aldose sugar glyceraldehyde. There is no way by which the structures of the two isomers of glyceraldehyde (see the formulas below, which are the so-called Fischer projection formulas that are commonly used to distinguish between such isomers) can be made identical, excluding breaking and reforming the linkages, or bonds, of the hydrogen (−H) and hydroxyl (−OH) groups attached to the carbon at position 2. The isomers are, in fact, mirror images akin to right and left hands; the term enantiomorphism is frequently employed for such isomerism. The chemical and physical properties of enantiomers are identical except for the property of optical (optical activity) rotation.

      As explained above, optical rotation is the rotation of the plane of polarized light. Polarized (polarization) light is light that has been separated into two beams that vibrate at right angles to each other; solutions of substances that rotate the plane of polarization are said to be optically active, and the degree of rotation is called the optical rotation of the solution. In the case of the isomers of glyceraldehyde, the magnitudes of the optical rotation are the same, but the direction in which the light is rotated—generally designated as plus, or d for dextrorotatory (to the right), or as minus, or l for levorotatory (to the left)—is opposite; i.e., a solution of D-(d)-glyceraldehyde causes the plane of polarized light to rotate to the right, and a solution of L-(l)-glyceraldehyde rotates the plane of polarized light to the left. Fischer projection formulas for the two isomers of glyceraldehyde are given below (see Configuration, below, for explanation of D and L).

Configuration

      Molecules, such as the isomers of glyceraldehyde—the atoms of which can have different structural arrangements—are known as asymmetrical molecules. The number of possible structural arrangements for an asymmetrical molecule depends on the number of centres of asymmetry; i.e., for n (any given number of) centres of asymmetry, 2ⁿ different isomers of a molecule are possible. An asymmetrical centre in the case of carbon is defined as a carbon atom to which four different groups are attached. In the three-carbon aldose sugar, glyceraldehyde, the asymmetrical centre is located at the central carbon atom. The four different groups attached to the atom are

      The position of the hydroxyl group (−OH) attached to the central carbon atom—i.e., whether −OH projects from the left or the right—determines whether the molecule rotates the plane of polarized light to the left, or to the right. Since glyceraldehyde has one asymmetrical centre, n is one in the relationship 2ⁿ, and there thus are two possible glyceraldehyde isomers. Sugars containing four carbon atoms have two asymmetrical centres; hence, there are four possible isomers (2²). Similarly, sugars with five carbon atoms have three asymmetrical centres, and thus have eight possible isomers (2³). Keto sugars have one less asymmetrical centre for a given number of carbon atoms than do aldehydo sugars.

 A convention of nomenclature, devised in 1906, states that the form of glyceraldehyde whose asymmetrical carbon atom has a hydroxyl group projecting to the right (see Fischer projection formulas) is designated as of the D-configuration; that form, whose asymmetrical carbon atom has a hydroxyl group projecting to the left, is designated as L. All sugars that can be derived from D-glyceraldehyde—i.e., hydroxyl group attached to the asymmetrical carbon atom most remote from the aldehydo or keto end of the molecule projects to the right—are said to be of the D-configuration; those sugars derived from L-glyceraldehyde are said to be of the L-configuration. See Table 1—> for aldoses—i.e., sugars containing an aldehydo group

of the D-configuration.

      The configurational notation D or L is independent of the sign of the optical rotation of a sugar in solution. It is common, therefore, to designate both, as, for example, D-(l)-fructose or D-(d)-glucose; i.e., both have a D-configuration at the centre of asymmetry most remote from the aldehydo end (in glucose) or keto end (in fructose) of the molecule, but fructose is levorotatory, and glucose is dextrorotatory—hence the latter has been given the alternative name dextrose. Although the initial assignments of configuration for the glyceraldehydes were made on purely arbitrary grounds, studies that were carried out nearly half a century later established them as correct in an absolute spatial sense. In biological systems, only the D or L form may be utilized.

 When more than one asymmetrical centre is present in a molecule, as is the case with sugars having four or more carbon atoms, a series of DL pairs exists, and they are functionally, physically, and chemically distinct; thus, although D-xylose and D-lyxose (see Table 1—>) both have five carbon atoms and are of the D-configuration, the spatial arrangement of the asymmetrical centres (at carbon atoms 2, 3, and 4) is such that they are not mirror images.

Hemiacetal and hemiketal forms

      Although optical rotation has been one of the most frequently determined characteristics of carbohydrates since its recognition in the late 19th century, the rotational behaviour of freshly prepared solutions of many sugars differs from that of solutions that have been allowed to stand. This phenomenon, termed mutarotation, is demonstrable even with apparently identical sugars and is caused by a type of stereoisomerism involving formation of an asymmetrical centre at the first carbon atom (aldehydo carbon) in aldoses and the second one (keto carbon) in ketoses.

 Most pentose and hexose sugars, therefore, do not exist as linear, or open-chain, structures in solution, as indicated for the aldoses in Table 1—>, but form cyclic, or ring, structures termed hemiacetal or hemiketal forms, respectively. As illustrated for glucose and fructose, the cyclic structures are formed by the addition of the hydroxyl group (−OH) from either the fourth, fifth, or sixth carbon atom (in the diagram, the numbers 1 through 6 represent the positions of the carbon atoms) to the carbonyl group

at position 1 in glucose or 2 in fructose. A five-membered ring is illustrated for the ketohexose, fructose; a six-membered ring is illustrated for the aldohexose, glucose. In either case, the cyclic forms are in equilibrium with (i.e., the rate of conversion from one form to another is stable) the open-chain structure—a free aldehyde if the solution contains glucose, a free ketone if it contains fructose; each form has a different optical rotation value. Since the forms are in equilibrium with each other, a constant value of optical rotation is measurable; the two cyclic forms represent more than 99.9 percent of the sugar in the case of a glucose solution.

      By definition, the carbon atom containing the aldehydo

is termed the anomeric carbon atom; similarly, carbohydrate stereoisomers that differ in configuration only at this carbon atom are called anomers. When a cyclic hemiacetal or hemiketal structure forms, the structure with the new hydroxyl group projecting on the same side as that of the oxygen involved in forming the ring is termed the alpha anomer (see hemiacetal forms for glucose and diagram); that with the hydroxyl group projecting on the opposite side from that of the oxygen ring is termed the beta anomer (see diagram).

      The spatial arrangements of the atoms in these cyclic structures are better shown (glucose is used as an example) in the representation devised by the British organic chemist Walter Norman (later Sir Norman) Haworth (Haworth, Sir Norman) about 1930; they are still in widespread use. In the formulation the asterisk indicates the position of the anomeric carbon atom; the carbon atoms, except at position 6, usually are not labelled.

      The large number of asymmetrical carbon atoms and the consequent number of possible isomers considerably complicates the structural chemistry of carbohydrates.

Classes of carbohydrates

Monosaccharides (monosaccharide)

Sources

       Some naturally occurring monosaccharidesThe most common naturally occurring monosaccharides are D-glucose, D-mannose, D-fructose, and D-galactose among the hexoses, and D-xylose and L-arabinose among the pentoses. In a special sense, D-ribose and 2-deoxy-D-ribose are ubiquitous because they form the carbohydrate component of ribonucleic acid (RNA) and deoxyribonucleic acid (DNA), respectively; these sugars are present in all cells as components of nucleic acids. Sources of some of the naturally occurring monosaccharides are listed in Table 2 (Some naturally occurring monosaccharides).

      D-xylose, found in most plants in the form of a polysaccharide called xylan, is prepared from corncobs, cottonseed hulls, or straw by chemical breakdown of xylan. D-galactose, a common constituent of both oligosaccharides and polysaccharides, also occurs in carbohydrate-containing lipids, called glycolipids, which are found in the brain and other nervous tissues of most animals. Galactose is generally prepared by acid hydrolysis (breakdown involving water) of lactose, which is composed of galactose and glucose. Since the biosynthesis of galactose in animals occurs through intermediate compounds derived directly from glucose, animals do not require galactose in the diet. In fact, in most human populations the majority of people do not retain the ability to manufacture the enzyme necessary to metabolize (metabolism) galactose after they reach the age of four, and many individuals possess a hereditary defect known as galactosemia and never have the ability to metabolize galactose.

      D-glucose (from the Greek word glykys, meaning “sweet”), the naturally occurring form, is found in fruits, honey, blood, and, under abnormal conditions, in urine. It is also a constituent of the two most common naturally found disaccharides, sucrose and lactose, as well as the exclusive structural unit of the polysaccharides cellulose, starch, and glycogen. Generally, D-glucose is prepared from either potato starch or cornstarch.

      D-fructose, a ketohexose, is one of the constituents of the disaccharide sucrose and is also found in uncombined form in honey, apples, and tomatoes. Fructose, generally considered the sweetest monosaccharide, is prepared by sucrose hydrolysis and is metabolized by humans.

Chemical reactions

      The reactions of the monosaccharides can be conveniently subdivided into those associated with the aldehydo or keto group and those associated with the hydroxyl groups.

      The relative ease with which sugars containing a free or potentially free aldehydo or keto group can be oxidized to form products has been known for a considerable time and once was the basis for the detection of these so-called reducing sugars in a variety of sources. For many years, analyses of blood glucose and urinary glucose were carried out by a procedure involving the use of an alkaline copper compound. Because the reaction has undesirable features—extensive destruction of carbohydrate structure occurs, and the reaction is not very specific (i.e., sugars other than glucose give similar results) and does not result in the formation of readily identifiable products—blood and urinary glucose now are analyzed by using the enzyme glucose oxidase, which catalyzes the oxidation of glucose to products that include hydrogen peroxide. The hydrogen peroxide then is used to oxidize a dye present in the reaction mixture; the intensity of the colour is directly proportional to the amount of glucose initially present. The enzyme, glucose oxidase, is highly specific for β-D-glucose.

      In another reaction, the aldehydo group of glucose

reacts with alkaline iodine to form a class

      of compounds called aldonic acids. One important aldonic acid is ascorbic acid (vitamin C, see structure), an essential dietary component for humans and guinea pigs. The formation of similar acid derivatives does not occur with the keto sugars.

      Either the aldehydo or the keto group of a sugar may be reduced (i.e., hydrogen added) to form an alcohol; compounds formed in this way are called alditols, or sugar alcohols. The product formed as a result of the reduction of the aldehydo carbon of D-glucose is called sorbitol (D-glucitol). D-glucitol also is formed when L-sorbose is reduced. The reduction of mannose results in mannitol, that of galactose in dulcitol.

      Sugar alcohols that are of commercial importance include sorbitol (D-glucitol), which is commonly used as a sweetening agent, and D-mannitol, which is also used as a sweetener, particularly in chewing gums, because it has a limited water solubility and remains powdery and granular on long storage.

Formation of glycosides

      The hydroxyl group that is attached to the anomeric carbon atom (i.e., the carbon containing the aldehydo or keto group) of carbohydrates in solution has unusual reactivity, and derivatives, called glycosides (glycoside), can be formed; glycosides formed from glucose are called glucosides. It is not possible for equilibration between the α- and β-anomers of a glycoside in solution (i.e., mutarotation) to occur. The reaction by which a glycoside is formed (see below) involves the hydroxyl group (−OH) of the anomeric carbon atom (numbered 1) of both α and β forms of D-glucose—α and β forms of D-glucose are shown in equilibrium in the reaction sequence—and the hydroxyl group of an alcohol (methyl alcohol in the reaction sequence); methyl α-D-glucosides and β-D-glucosides are formed as products, as is water.

      Among the wide variety of naturally occurring glycosides are a number of plant pigments (pigment), particularly those red, violet, and blue in colour (coloration); these pigments are found in flowers and consist of a pigment molecule attached to a sugar molecule, frequently glucose. Plant indican (from Indigofera species), composed of glucose and the pigment indoxyl, was important in the preparation of indigo dye before synthetic dyes became prevalent. Of a number of heart-muscle stimulants that occur as glycosides, digitalis is still used. Other naturally occurring glycosides include vanillin, which is found in the vanilla bean, and amygdalin (oil of bitter almonds); a variety of glycosides found in mustard have a sulfur atom at position 1 rather than oxygen.

      A number of important antibiotics are glycosides; the best known are streptomycin and erythromycin. Glucosides—i.e., glycosides formed from glucose—in which the anomeric carbon atom (at position 1) has phosphoric acid linked to it, are extremely important biological compounds.

      For example, α-D-glucose-1-phosphate (see formula), is an intermediate product in the biosynthesis of cellulose, starch, and glycogen; similar glycosidic phosphate derivatives of other monosaccharides participate in the formation of naturally occurring glycosides and polysaccharides.

      The hydroxyl groups other than the one at the anomeric carbon atom can undergo a variety of reactions, several of which deserve mention. Esterification (ester), which consists of reacting the hydroxyl groups with an appropriate acidic compound, results in the formation of a class of compounds called sugar esters. Among the common ones are the sugar acetates, in which the acid is acetic acid. Esters of phosphoric acid and sulfuric acid are important biological compounds; glucose-6-phosphate, for example, plays a central role in the energy metabolism of most living cells, and D-ribulose 1,5-diphosphate is important in photosynthesis.

Formation of methyl ethers

      Treatment of a carbohydrate with methyl iodide or similar agents under appropriate conditions results in the formation of compounds in which the hydroxyl groups are converted to methyl groups (−CH₃). Called methyl ethers, these compounds are employed in structural studies of oligosaccharides and polysaccharides because their formation does not break the bonds, called glycosidic bonds, that link adjacent monosaccharide units. In the reaction sequence shown, a segment of a starch molecule, consisting of three glucose units, is indicated; the Haworth formulation used to represent one of the glucose units shows the locations of the glycosidic bonds and the −OH groups. When complete etherification of the starch molecule is carried out, using methyl iodide, methyl groups become attached to the glucose molecules at the three positions shown in the methylated segment of the starch molecule; note that the glycosidic bonds have not been broken by the reaction with methyl iodide. When the methylated starch molecule then is broken down (hydrolyzed), hydroxyl groups are located at the positions in the molecule previously involved in linking one sugar molecule to another, and a methylated glucose, in this case named 2,3,6 tri-O-methyl-D-glucose, forms. The linkage positions (in the example, at carbon atoms 1 and 4; the carbon atoms are numbered in the structure of the methylated glucose), which are not methylated, in a complex carbohydrate can be established by analyzing the locations (in the example, at carbon atoms 2, 3, and 6) of the methyl groups in the monosaccharides. This technique is useful in determining the structural details of polysaccharides, particularly since the various methylated sugars are easily separated by techniques involving gas chromatography, in which a moving gas stream carries a mixture through a column of a stationary liquid or solid, the components thus being resolved.

      When the terminal group (CH₂OH) of a monosaccharide is oxidized chemically or biologically, a product called a uronic acid is formed. Glycosides that are derived from D-glucuronic acid (the uronic acid formed from D-glucose) and fatty substances called steroids appear in the urine of animals as normal metabolic products; in addition, foreign toxic substances are frequently converted in the liver to glucuronides before excretion in the urine. D-glucuronic acid also is a major component of connective tissue polysaccharides, and D-galacturonic acid and D-mannuronic acid, formed from D-galactose and D-mannose, respectively, are found in several plant sources.

      Other compounds formed from monosaccharides include those in which one hydroxyl group, usually at the carbon at position 2 (see formulas for D-glucosamine and D-galactosamine), is replaced by an amino group (−NH₂); these compounds, called amino sugars, are widely distributed in nature. The two most important ones are glucosamine (2-amino-2-deoxy-D-glucose) and galactosamine (2-amino-2-deoxy-D-galactose).

      Neither amino sugar is found in the uncombined form. Both occur in animals as components of glycolipids or polysaccharides; e.g., the primary structural polysaccharide (chitin) of insect outer skeletons and various blood-group substances.

      In a number of naturally occurring sugars, known as deoxy sugars, the hydroxyl group at a particular position is replaced by a hydrogen atom. By far the most important representative is 2-deoxy-D-ribose (see formula), the pentose sugar found in deoxyribonucleic acid (DNA); the hydroxyl group at the carbon atom at position 2 has been replaced by a hydrogen atom.

      Other naturally occurring deoxy sugars are hexoses, of which L-rhamnose (6-deoxy-L-mannose) and L-fucose (6-deoxy-L-galactose) are the most common; the latter, for example, is present in the carbohydrate portion of blood-group substances and in red-blood-cell membranes.

Disaccharides (disaccharide) and oligosaccharides

      Disaccharides are a specialized type of glycoside in which the anomeric hydroxyl group of one sugar has combined with the hydroxyl group of a second sugar with the elimination of the elements of water. Although an enormous number of disaccharide structures are possible, only a limited number are of commercial or biological significance.

Sucrose and trehalose

      Sucrose, or common table sugar, has a world production amounting to well over 10,000,000 tons annually. The unusual type of linkage between the two anomeric hydroxyl groups of glucose and fructose (see formula, in which the asterisk indicates anomeric carbon atom) means that neither a free aldehydo group (on the glucose moiety) nor a free keto group (on the fructose moiety) is available to react unless the linkage between the monosaccharides is destroyed; for this reason, sucrose is known as a nonreducing sugar. Sucrose solutions do not exhibit mutarotation, which involves formation of an asymmetrical centre at the aldehydo or keto group. If the linkage between the monosaccharides composing sucrose is broken, the optical rotation value of sucrose changes from positive to negative; the new value reflects the composite rotation values for D-glucose, which is dextrorotatory (+52°), and D-fructose, which is levorotatory (−92°). The change in the sign of optical rotation from positive to negative is the reason sucrose is sometimes called invert sugar.

      The commercial preparation of sucrose takes advantage of the alkaline stability of the sugar, and a variety of impurities are removed from crude sugarcane extracts by treatment with alkali. After this step, syrup preparations are crystallized to form table sugar. Successive “crops” of sucrose crystals are “harvested,” and the later ones are known as brown sugar. The residual syrupy material is called either cane final molasses or blackstrap molasses; both are used in the preparation of antibiotics, as sweetening agents, and in the production of alcohol by yeast fermentation.

      Sucrose is formed following photosynthesis in plants by a reaction in which sucrose phosphate first is formed.

      The disaccharide trehalose is similar in many respects to sucrose but is much less widely distributed. It is composed of two molecules of α-D-glucose and is also a nonreducing sugar. Trehalose is present in young mushrooms and in the resurrection plant (Selaginella); it is of considerable biological interest because it is also found in the circulating fluid (hemolymph) of many insects. Since trehalose can be converted to a glucose phosphate compound by an enzyme-catalyzed reaction that does not require energy, its function in hemolymph may be to provide an immediate energy source, a role similar to that of the carbohydrate storage forms (i.e., glycogen) found in higher animals.

lactose and maltose

      Lactose is one of the sugars (sucrose is another) found most commonly in human diets throughout the world; it composes about 5 percent or more of the milk of all mammals. Lactose consists of two aldohexoses—β-D-galactose and glucose—linked so that the aldehydo group at the anomeric carbon of glucose is free to react (see structural formula, in which the asterisk indicates position of anomeric carbon atoms); i.e., lactose is a reducing sugar.

      A variety of metabolic disorders related to lactose may occur in infants; in some cases, they are the result of a failure to metabolize properly the galactose portion of the molecule.

 Although not found in uncombined form in nature, the disaccharide maltose is biologically important because it is a product of the enzymatic breakdown of starches during digestion. Maltose consists of α-D-glucose linked to a second glucose unit in such a way that maltose is a reducing sugar. Maltose, which is readily hydrolyzed to glucose and can be metabolized by animals, is employed as a sweetening agent and as a food for infants whose tolerance for lactose is limited. Table 3—> lists the component sugars of, the linkage between, and the occurrence of a number of disaccharides and oligosaccharides.

Polysaccharides (polysaccharide)

 Polysaccharides, or glycans, may be classified in a number of ways; the following scheme is frequently used. Homopolysaccharides are defined as polysaccharides formed from only one type of monosaccharide. Homopolysaccharides may be further subdivided into straight-chain and branched-chain representatives, depending upon the arrangement of the monosaccharide units. Heteropolysaccharides are defined as polysaccharides containing two or more different types of monosaccharides; they may also occur in both straight-chain and branched-chain forms. In general, extensive variation of linkage types (see Table 4—>) does not occur within a polysaccharide structure, nor are there many polysaccharides composed of more than three or four different monosaccharides; most contain one or two.

Homopolysaccharides

 In general, homopolysaccharides have a well-defined chemical structure, although the molecular weight of an individual amylose or xylan molecule may vary within a particular range, depending on the source; molecules from a single source also may vary in size, because most polysaccharides are formed biologically by an enzyme-catalyzed process lacking genetic information regarding size. Several naturally occurring homopolysaccharides are listed in Table 4—>.

      The basic structural component of most plants, cellulose, is widely distributed in nature. It has been estimated that nearly 10,000,000,000 tons of cellulose are synthesized yearly as a result of photosynthesis by higher plants. The proportion of cellulose to total carbohydrate found in plants may vary in various types of woods from 30 to 40 percent, and to more than 98 percent in the seed hair of the cotton plant. Cellulose, a large, linear molecule composed of 3,000 or more β-D-glucose molecules, is insoluble in water.

      The chains of glucose units composing cellulose molecules are frequently aligned within the cell-wall structure of a plant to form fibre-like or crystalline arrangements. This alignment permits very tight packing of the chains and promotes their structural stability but also makes structural analysis difficult. The relationships between cellulose and other polysaccharides present in the cell wall are not well established; in addition, the presence of unusual chemical linkages or nonglucose units within the cellulose structure has not yet been established with certainty.

      During the preparation of cellulose, raw plant material is treated with hot alkali; this treatment removes most of the lignin, the hemicelluloses, and the mucilaginous components. The cellulose then is processed to produce papers and fibres. The high resistance of cellulose to chemical or enzymatic breakdown is important in the manufacture of paper and cloth. Cellulose also is modified chemically for other purposes; e.g., compounds such as cellulose acetate are used in the plastics industry, in the production of photographic film, and in the rayon-fibre industry. Viscose rayon is produced from an ester of cellulose, and cellulose nitrate is employed in the lacquer and explosives industries.

      The noteworthy biological stability of cellulose is dramatically illustrated by trees, the life-span of which may be several thousand years. Enzymes capable of breaking down cellulose are generally found only among several species of bacteria and molds. The apparent ability of termites to utilize cellulose as an energy source depends on the presence in their intestinal tracts of protozoans that can break it down. Similarly, the single-celled organisms present in the rumina of sheep and cattle are responsible for the ability of these animals to utilize the cellulose present in typical grasses and other feeds.

 Xylans are almost as ubiquitous as cellulose in plant-cell walls and contain predominantly β-D-xylose units linked as in cellulose (see Table 4—>). Some xylans contain other sugars, such as L-arabinose, but they form branches and are not part of the main chain. Xylans are of little commercial importance.

      The term starch refers to a group of plant reserve polysaccharides consisting almost exclusively of a linear component (amylose) and a branched component (amylopectin). The use of starch as an energy source by humans depends on the ability to convert it completely to individual glucose units; the process is initiated by the action of enzymes called amylases, synthesized by the salivary glands in the mouth, and continues in the intestinal tract. The primary product of amylase action is maltose, which is hydrolyzed to two component glucose units as it is absorbed through the walls of the intestine.

      A characteristic reaction of the amylose component of starch is the formation with iodine of a complex compound with a characteristic blue colour. About one iodine molecule is bound for each seven or eight glucose units, and at least five times that many glucose units are needed in an amylose chain to permit the effective development of the colour.

      The amylopectin component of starch is structurally similar to glycogen in that both are composed of glucose units linked together in the same way, but the distance between branch points (see schematic diagrams, in which −O− represents one glucose unit) is greater in amylopectin than in glycogen, and the former may be thought of as occupying more space per unit weight.

      The applications of starches other than as foods are limited. Starches are employed in adhesive manufacture, and starch nitrate has some utility as an explosive.

      Glycogen, which is found in all animal tissues, is the primary animal storage form of carbohydrate and, indirectly, of rapidly available energy. The distance between branch points in a glycogen molecule is only five or six units (see schematic diagram above), which results in a compact treelike structure. The ability of higher animals to form and break down this extensively branched structure is essential to their well-being; in conditions known as glycogen storage diseases, these activities are abnormal, and the asymmetrical glycogen molecules that are formed have severe, often fatal, consequences. Glycogen synthesis and breakdown are controlled by substances called hormones.

      Large molecules—e.g., pectins (pectin) and agars—composed of galactose or its uronic-acid derivative (galacturonic acid) are important because they can form gels. Pectins, which are predominantly galacturonans, are produced from citrus fruit rinds; they are used commercially in the preparation of jellies and jams. Agar is widely employed in biological laboratories as a solidifying agent for growth media for microorganisms and in the bakery industry as a gelling agent; it forms a part of the diet of people in several areas of East Asia.

      Dextrans, a group of polysaccharides composed of glucose, are secreted by certain strains of bacteria as slimes. The structure of an individual dextran varies with the strain of microorganism. Dextrans can be used as plasma expanders (substitutes for whole blood) in cases of severe shock. In addition, a dextran derivative compound is employed medically as an anticoagulant for blood.

       chitin is structurally similar to cellulose, but the repeating sugar is 2-deoxy-2-acetamido-D-glucose (N-acetyl-D-glucosamine, see structural formula) rather than glucose.

      Sometimes referred to as animal cellulose, chitin is the major component of the outer skeletons of insects, crustaceans, and other arthropods, as well as annelid and nematode worms, mollusks, and coelenterates. The cell walls of most fungi also are predominantly chitin, which comprises nearly 50 percent of the dry weight of some species. Since chitin is nearly as chemically inactive as cellulose and easily obtained, numerous attempts, none of which has thus far been successful, have been made to develop it commercially. The nitrogen content of the biosphere, however, is stabilized by the ability of soil microorganisms to degrade nitrogen-containing compounds such as those found in insect skeletons; these microorganisms convert the nitrogen in complex molecules to a form usable by plants. If such microorganisms did not exist, much of the organic nitrogen present in natural materials would be unavailable to plants.

Heteropolysaccharides

       Representative heteropolysaccharidesIn general, heteropolysaccharides (heteroglycans) contain two or more different monosaccharide units. Although a few representatives contain three or more different monosaccharides, most naturally occurring heteroglycans contain only two different ones and are closely associated with lipid or protein. The complex nature of these substances has made detailed structural studies extremely difficult. The major heteropolysaccharides include the connective-tissue polysaccharides, the blood-group substances, glycoproteins (combinations of carbohydrates and proteins) such as gamma globulin, and glycolipids (combinations of carbohydrates and lipids), particularly those found in the central nervous system of animals and in a wide variety of plant gums (see Table 5 (Representative heteropolysaccharides)).

      The most important heteropolysaccharides are found in the connective tissues of all animals and include a group of large molecules that vary in size, shape, and interaction with other body substances. They have a structural role, and the structures of individual connective-tissue polysaccharides are related to specific animal functions; hyaluronic acid, for example, the major component of joint fluid in animals, functions as a lubricating agent and shock absorber.

       Representative heteropolysaccharidesThe connective-tissue heteropolysaccharides contain acidic groups (uronic acids or sulfate groups) and can bind both water and inorganic metal ions. They can also play a role in other physiological functions; e.g., in the accumulation of calcium before bone formation. Ion-binding ability also appears to be related to the anticoagulant activity of the heteropolysaccharide heparin (see Table 5 (Representative heteropolysaccharides)).

      The size of the carbohydrate portion of glycoproteins such as gamma globulin or hen-egg albumin is usually between five and 10 monosaccharide units; several such units occur in some glycoprotein molecules. The function of the carbohydrate component has not yet been established except for glycoproteins associated with cell surfaces; in this case, they appear to act as antigenic determinants—i.e., they are capable of inducing the formation of specific antibodies.

Preparation and analysis

      In general, monosaccharides are prepared by breakdown with acids of the polysaccharides in which they occur. Sugars usually are difficult to obtain in crystalline form, and the crystallization process usually is begun by “seeding” a concentrated solution of the sugar with crystals. The techniques employed for separation of monosaccharides depend to some extent on their physical and chemical properties; chromatographic procedures are often used.

      Oligosaccharides (oligosaccharide) and polysaccharides are prepared from natural sources by techniques that take advantage of size, alkaline stability, or some combination of these and other properties of the molecule of interest. It should be noted that preparation of an oligosaccharide or polysaccharide usually results in a range of molecular sizes of the desired molecule. The purity of a carbohydrate preparation, which is frequently based on an analysis of its composition, is more easily established for monosaccharides and disaccharides than for large, insoluble molecules such as cellulose.

Analytical techniques

      A variety of organic chemical analytical techniques are generally applicable to studies involving carbohydrates. Optical rotation, for example, once was frequently used to characterize carbohydrates. The ability to measure the rotation of the plane of polarized light transmitted through a solution containing a carbohydrate depends on finding a suitable solvent; water usually is used, with light at a wavelength of 589 mμ (millimicrons). Optical rotation is no longer widely used to characterize monosaccharides. The magnitude and sign of the optical rotation of glycosides, however, is useful in assigning configuration (α or β) to the hydroxyl group at the anomeric centre; glycosides of the α-configuration generally have rotations of higher magnitude than do the same glycosides of the β-configuration. Optical rotation is not a completely additive property; a trisaccharide composed of three glucose residues, for example, does not have a rotation three times that of one glucose molecule. Sugar alcohols cannot form ring structures; their rotation values are extremely small, suggesting a relationship between ring structure and the ability of a carbohydrate to rotate the plane of polarized light. Certain types of reactions (e.g., glycoside hydrolysis) can be monitored by measuring the change in optical rotation as a function of time. This technique is frequently used to examine the breakdown of disaccharides or oligosaccharides to monosaccharide units, especially if a large change in the net optical rotation may be expected, as occurs in the hydrolysis of sucrose.

Spectroscopic techniques

      Several other optical techniques used in chemistry have been applied to the analysis of carbohydrates. Infrared spectroscopy, used to measure vibrational and rotational excitation of molecules, and nuclear magnetic-resonance spectroscopy, which measures the excitation of certain components of molecules in a magnetic field induced by radio-frequency radiation, are valuable, although the similarity of the functional groups (i.e., the hydroxyl groups) limits use of the former technique for most sugars. Proton magnetic-resonance spectroscopy, nuclear magnetic resonance applied to protons (H atoms), is employed to identify the relative spatial arrangements of individual hydrogen atoms in a molecule. When they are precisely placed, the corresponding positions of the hydroxyl groups attached to the same carbon atom can be deduced. An extension of this technique utilizes the resonance spectroscopy of carbon-13, a nonradioactive isotope of carbon, so that ring structures can be established with great accuracy. Both the proton and carbon magnetic resonance methods are best applied to monosaccharides; they are less valuable in studying polysaccharides because an individual hydrogen atom in a large molecule is too small for accurate detection.

Identification of subunits

      The study of polysaccharide structure usually focuses on the chemical composition, the linkage between the monosaccharide units, and the size and shape of the molecule. The size and shape of a polysaccharide can be ascertained by techniques that are usually applied to large molecules; e.g., the most accurate molecular weight determination measures the sedimentation properties of the molecule in an applied gravitational field (e.g., the rate at which a solid material is deposited from a state of suspension or solution in a liquid). Indications of the shape of polysaccharide molecules in solution are obtained from viscosity measurements, in which the resistance of the molecules to flow (viscosity) is equated with the end-to-end length of the molecule; the viscosity of hyaluronic acid, for example, shows a marked dependence on both concentration of the acid and the salt content of the solution, and, under conditions approximating those found in biological systems, a hyaluronic acid molecule may be thought of as occupying a great deal of space. Alternatively, the compact nature of a glycogen molecule of molecular weight equal to that of a molecule of hyaluronic acid results in its accommodation to a much smaller space than the latter molecule.

      The identification of sugars in a mixture resulting from the hydrolytic breakdown of a heteropolysaccharide is most often carried out by chromatography of the mixture on paper, silica gel, or cellulose. Ready separations can be achieved between pentoses, hexoses and, for example, deoxy sugars; closely related compounds such as D-glucose and D-galactose also can be separated using chromatographic techniques. The linkage positions in polysaccharides are usually determined using the methylation procedure described previously. The various monosaccharide methyl ethers produced by the methylation are separated by gas–liquid chromatography.

      Detailed statements about polysaccharide structure and function are limited by the statistical nature of some measurements (e.g., branching frequency), the biological variability of parameters such as size and molecular weight, and incomplete information about associative interactions in living things.

Eugene A. Davidson

Additional Reading

Comprehensive works are Henry R. Mahler and Eugene H. Cordes, Basic Biological Chemistry (1968); Abraham White et al., Principles of Biochemistry, 6th ed. (1978); Albert L. Lehninger, David L. Nelson, and Michael L. Cox, Principles of Biochemistry, 2nd ed. (1993); Thomas Briggs and Albert M. Chandler (eds.), Biochemistry, 2nd ed. (1992); John W. Hill, Dorothy M. Feigl, and Stuart J. Baum, Chemistry and Life, 4th ed. (1993); Lubert Stryer, Biochemistry, 3rd ed. (1988); Donald Voet and Judith G. Voet, Biochemistry (1990); Geoffrey Zubay, Biochemistry, 3rd ed. (1993); and Laurence A. Moran et al., Biochemistry, 2nd ed. (1994). David J. Holme and Hazel Peck, Analytical Biochemistry, 2nd ed. (1993), covers newer methods of analysis. Ed.Studies focusing specifically on carbohydrates include S.F. Dyke, The Carbohydrates (1960), an introduction to basic reactions in the carbohydrate field, written almost in outline, schematic form; R.D. Guthrie and John Honeyman, An Introduction to the Chemistry of Carbohydrates, 3rd ed. (1968), a short monograph on basic carbohydrate structure and reactions; R.J. McIlroy, Introduction to Carbohydrate Chemistry (1967), a short introduction to carbohydrate chemistry recognizing the importance of stereochemistry and conformational factors; E.G.V. Percival, Structural Carbohydrate Chemistry, 2nd ed., rev. by Elizabeth Percival (1962), a standard work detailing methods of structural determination, particularly of sugar ring size and polysaccharide structure, still useful as a summary of structural methods even though many modern techniques are not discussed; Ward Pigman, Derek Horton, and Anthony Herp (eds.), The Carbohydrates: Chemistry and Biochemistry, 2nd ed., 2 vol. in 4 (1970–80), a standard reference work; Jaroslav Staněk, The Monosaccharides, trans. from Czech (1963); and Jaroslav Staněk, Miloslav Černý, and Josef Pačak, The Oligosaccharides, trans. from Czech (1965), standard reference works detailing structure, reaction, and properties of monosaccharides and oligosaccharides; Roy L. Whistler and Charles Louis Smart, Polysaccharide Chemistry (1953), a summary of chemical information on polysaccharides up to 1952, with emphasis on homopolymers such as starch and cellulose since few detailed structures were known at that time; Eugene A. Davidson, Carbohydrate Chemistry (1967), an intermediate college-level text emphasizing stereochemistry, conformation, and modern organic reaction mechanisms as applied to carbohydrates, including both physical and chemical methods for structural determination; Marcel Florkin and Elmer H. Stotz (eds.), Comprehensive Biochemistry, vol. 5, Carbohydrates (1963), a detailed volume covering all aspects of monosaccharide and many areas of polysaccharide structure and chemistry, although physical methods do not receive sufficient attention; M. Stacey and S.A. Barker, Carbohydrates of Living Tissues (1962), a detailed account of the chemistry and biochemistry of polysaccharide substances, primarily those found in animal tissues; and Karla L. Roehrig, Carbohydrate Biochemistry and Metabolism (1984).Works primarily of historical interest are Frederick J. Bates et al., Polarimetry, Saccarimetry and the Sugars (1942), practical information on sucrose and other common sugars together with methods for analysis and preparation of simple derivatives—many of the data tables provide useful reference information; and Walter Norman Haworth, The Constitution of the Sugars (1929), a classic description of the knowledge of sugar chemistry at that time, particularly Haworth's work in defining the ring structure of the carbohydrates.Methods of analysis are described in Roy L. Whistler and M.L. Wolfrom (eds.), Methods in Carbohydrate Chemistry (1962– ), a necessary reference work for research workers in the field, detailing laboratory procedures for monosaccharide and polysaccharide preparations; M.F. Chaplin and John F. Kennedy (eds.), Carbohydrate Analysis: A Practical Approach (1986); and two works dealing with carbohydrate analysis using nuclear magnetic resonance, gas chromatography, and mass spectrometry: Christopher J. Biermann and Gary D. McGinnis (eds.), Analysis of Carbohydrates by GLC and MS (1989); and Lawrence J. Berliner and Jacques Reuben (eds.), Carbohydrates and Nucleic Acids (1992). John F. Kennedy (ed.), Carbohydrate Chemistry (1988); and J. Thiem (ed.), Carbohydrate Chemistry (1990), include information on synthesis.Ongoing research is published in Advances in Carbohydrate Chemistry and Biochemistry (annual), multiauthored volumes reviewing specific areas of carbohydrate chemistry and biology. Eugene A. Davidson Ed.

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