XII. POLYSACCHARIDES (7) Parti General Aspects and Phyto and Microbial Polysaccharides ROY L. WHISTLER AND W. M. CORBETT Polysaccharides are high-molecular-weight carbohydrates. They may be viewed as condensation polymers in which monosaccharides (or their deriv- atives such as the uronic acids and aminosugars) have been glycosidically joined with the elimination of water according to the empirical equation: n C6H12Oe -+ [CeH10O6]„ + (n - 1)H20 From the reverse direction this equation states that polysaccharides, on complete hydrolysis, yield only simple sugars (or their derivatives). Poly- saccharides and oligosaccharides (Chapter IX) are thus, to this extent, defined similarly. However, the term polysaccharide is limited generally to those monosaccharide condensation polymers which contain ten or more monosaccharide residues. As was seen in Chapter IX, oligosaccharides contain two to nine sugar residues. Obviously this limitation of the two terms is completely arbitrary. It is justified on the basis that, as found in nature, most oligosaccharides contain two or three monosaccharide units in contrast to most polysaccharides which contain a hundred to several thousand monosaccharide units. Carbohydrates containing 5 to 15 sugar residues have not been found in nature although they surely occur, at least in small amounts. A few natural polysaccharides contain 30 to 100 sugar residues but most contain more. 1. NOMENCLATURE AND CLASSIFICATION In the early periods of carbohydrate chemistry no systematic nomen- clature existed. A polysaccharide name coined at that time usually re- flected the origin of the polysaccharide or sometimes emphasized some property of the isolated substance. Illustrative examples of such poly- saccharide nomenclature are found in the terms cellulose, the principal 1. For a comprehensive review of polysaccharide chemistry see R. L. Whistler and C. L. Smart, "Polysaccharide Chemistry." Academic Press, New York, 1953. 641 642 R. L. WHISTLER AND W. M. CORBETT component of cell walls in plants, and starch, a name derived from the Anglo-Saxon "stercan" meaning to stiffen. Names like these were not de- signed with a view toward their being put into a systematic nomenclature. Yet they have been used so extensively that they must be accepted in spite of their deficiencies. Progress toward a systematic nomenclature has produced the significant ending -an to designate that a substance is a polysaccharide. (The ending -osan signifies a simple sugar anhydride (see Chapter VII).) Thus, another word for polysaccharide is the generic term glycan. This term is evolved from the generic word glycose, meaning a simple sugar, and the ending -an, signifying a sugar polymer. Although not all older polysaccharide names employ this ending, it is found generally in names of polysaccharides composed of one or two sugar types: araban for an arabinose polymer, xylan for polymers of xylose, mannan for those of mannose, galactan for those of galactose, and galactomannans for galactose-mannose combina- tions. Until recently certain polymers of fructose were called fructosans. The name of the fructose polymers has been correctly shortened to fruc- tans. Persistent because of its wide use in the literature is the term pentosan, which designates polysaccharides composed of pentose sugar residues. This obviously unsystematic term has so far not been shortened to pentan or glycopentan. Cellulosan as a term for polysaccharides closely associated with cellulose is little used in modern carbohydrate literature. Many polysaccharide names ending with the unsystematic and un- desirable -in ending have been changed to end in -an as a step toward uniformity. Such terms are laminaran, carrageenan, lichenan, isolichenan, asparagan, senistran, graminan, tritican, kritesan, phlean, secalan, poan, pyrosan, and irisan. Names which have not been changed because of their long-standing and wide use are: pectin, amylopectin, inulin, chitin, heparin, and chondroitin. The systematic name glucan does not refer to a specific polysaccharide but signifies only that the polysaccharide is composed of glucose residues. The manner of Unkage and the arrangement are not specified. The name is a group name only and applies as well to cellulose as to glycogen, laminaran, or other glucose polymers. The polysaccharide can be defined more specifi- cally if a source designation is also employed as part of the name. Thus, a more definite polysaccharide is specified in each case by the designation beechwood xylan, yeast mannan, or peanut araban. Polysaccharides of the same type differ at least slightly from one source to another. Sometimes the differences are quite marked, as with starches. There is a well-known and readily apparent difference among starches xii. POLYSACCHARIDES: PART I 643 from different plants. A particular starch is meant, however, by banana starch or corn starch. Ideally, the polysaccharides should be classified according to their chemical composition and structure as has been suggested by Whistler (1). In such a classification, polysaccharides hydrolyzing to only one mono- saccharide type are termed homoglycans while polysaccharides hydrolyz- ing to two or three or more monosaccharide types are termed heteroglycans with prefixes of di-, tri-, and so on to designate the number of different types of sugar units. At present there is no proof that more than five or six types of sugar units occur in a single polysaccharide. The number of types of sugar units contained in a polysaccharide can easily be determined by Chromatographie examination of the hydrolyzate, provided, of course, that the polysaccharide is pure. In this structural classification the first logical subclassification separates polysaccharides as to whether they are linear or branched. This separation can readily be made by performing several simple tests. The easiest test is that of film formation. An aqueous polysaccharide solution when spread on a glass plate and dried will be brittle if a branched polysaccharide is present. Films from linear molecules will be strong, undergo folding without breaking and when plasticized can be stretched with the development of birefringence and a detectable "fi- brous" X-ray pattern. Linear polysaccharides also show streaming bire- fringence when their solutions are stirred and viewed between crossed polarizing plates. Methylation studies may further reveal whether a mole- cule is branched. Unfortunately, many polysaccharides have not been examined in rigor- ously pure conditions, and often examination has not extended to charac- terization of films. Therefore, at times, it is useful to classify polysaccharides according to source. In subsequent discussion, the polysaccharides will be discussed from the standpoint of their sources as: Phytopolysaccharides (Phytoglycans) Bacterial and Fungal Polysaccharides Zoöpolysaccharides (Zoöglycans) and Conjugates (Glycoproteins and Glycolipides) The phytopolysaccharides and the bacterial and fungal polysaccharides will be considered in Part I of this chapter, and the zoöpolysaccharides in Part II. This classification, however, has the definite disadvantage that some polysaccharides of identical or similar structure are separated ac- cording to their origin. Thus, amylopectin (plants) and glycogen (animals) are separated. Chitin is found in lower animals, microorganisms, and fungi. Cellulose, although considered typical of plants, is also produced by some bacteria and lower animals. 644 K. L. WHISTLER AND W. M. CORBETT 2. THE STRUCTURES OF POLYSACCHARIDES A. GENERAL CONCEPTS Though polysaccharides may be viewed as condensation polymers formed by the combination of monosaccharides with the elimination of the elements of water, the naturally occurring polysaccharides are far less complicated than would occur if the combination of monomers took place in random fashion. In fact many simplifying features are apparent on careful examination of all known polysaccharide structures. The basic reasons for such simplified and ordered arrangements stem from the action of those specific synthesizing enzymes which produce the monosaccharides and those enzymes which connect the monosaccharides, by various and sometimes complex routes, to polymer structures. Methods by which enzymes produce polysaccharides are given on page 703. In the condensation of monosaccharides to form natural polymers, the hydroxyl on the anomeric carbon always participates in the condensation. Since most polysaccharides are composed of aldose sugar units, this dis- cussion will be confined to such units. The hydroxyl on carbon atom 1 (anomeric carbon) may condense with any hydroxyl other than that at C-l on an adjoining monosaccharide unit. In this way, a linear chain can be formed with a free C-l hydroxyl group at one end. A complete random- ness of linkage with the various hydroxyl groups has never been found in nature. Most frequently, a particular mode of linkage is repeated uniformly through the chain. Even the stereoconfiguration of C-l remains constant in most observed cases. Thus, in amylose, there is a uniform α-Ό-1—»4 link- age, in cellulose a uniform β-Ό-1—*4 linkage, and in laminaran an essen- tially uniform β-D-l—>3 linkage. A further and highly simplifying fact in polysaccharide chemistry is that of the multitude of stereoisomeric monosaccharides only a very few are found in natural polysaccharides. Of the hexoses there are D-glucose, D-man- nose, D-fructose, D-galactose, and infrequently L-galactose, and possibly D-idose or L-altrose. Of