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REVIEW Seed Galactomannans: An Overview

by Manjoosha Srivastava and V. P. Kapoor* Pharmacognosy, Ethnopharmacology, and Phyochemistry Divisions, National Botanical Research Institute (NBRI), Lucknow-226001, India (e-mail: sharad_ks2003@yahoo.co.in)

Seed galactomannans are vegetable, heterogeneous polysaccharides widely distributed in nature. Generally, they possess (1 ! 4)-linked d-mannopyranose (Man) main chains to which are attached (1 ! 6)linked d-galactopyranosyl (Gal) units. The Man/Gal ratios differ from gum to gum, resulting in a change in structure, which, in turn, determines the various industrial applications of seed galactomannans. These materials are important in paper, textile, petroleum-drilling, pharmaceutics, food, cosmaceutics, and explosives industries. In this review, the biodiversified applicability of galactomannan gums is discussed, particularly with respect to structural aspects, properties, human consumption, and technical applications. Especially important is that the solution properties (rheological behaviour, viscosity, emulsifying tendency, etc.) of natural and chemically modified galactomannans can be tuned by interaction with other (carbohydrate-based) monomers or polymers.

1. Introduction. Galactomannans have attracted considerable academic attention as well as industrial interest due to their property of forming viscous solutions or gels in aqueous media. Seed galactomannans are heterogeneous polysaccharides widely distributed in nature. Generally, they possess main chains made of (1 ! 4)-linked dmannopyranose (Man) units to which are attached (1 ! 6)-linked d-galactopyranose (Gal) units. According to Aspinall [1], galactomannans are those mannans that contain more than 5% of galactose. Galactomannan gums can often be used in different forms for human consumption. The gums differ from each other by different Man/Gal ratios. The different chemical properties of these gums make them versatile materials used for many different applications. Some important properties are holding of H2O, thickening, gelling, binding, suspending, emulsifying, as well as formation of films. These characteristics have opened avenues to be utilized in various industries such as paper, textiles, petroleum, drilling, pharmaceutics, food, cosmaceutics, explosives, etc. The properties of gums can be enhanced by interaction with other monomers and polymers, mainly due to their capabilities to form synergistic interactions due to the numerous OH groups present. Carob gum forms gels at higher concentration, but guar does not possess gelling properties, although it forms viscous solutions. Interaction of carob with little of xanthan produces a gelling for commercial application. The scope to exploit seed gums obtained from different sources for varied applications is very large. There are numerous patents on seed-gum applications in
 2005 Verlag Helvetica Chimica Acta AG, Zrich

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developed countries [2]. The importance of these materials has strongly increased because of the global trend to use more plant-based products for ecological reasons. Plant-based products are considered GRAS (generally recognised as safe) [3] and ecologically friendly, as their production and application neither cause any pollution nor disturb the ecosystem. Moreover, these materials are inexpensive. Hence, for the study of the biodiversified application of gums, a detailed review, including chemical, structural, supramolecular, and rheological aspects of seed galactomannans, was considered important. 2. Important Sources of Galactomannans. Seed gums are mostly obtained from leguminous plants (Table 1), the endosperms of the seeds being mainly responsible for water-solubility [4 33]. The two main groups of galactomannan polysaccharides are derived either from the endosperm of plant seeds (the vast majority originating from leguminosae) or from microbial sources, in particular yeast [27] [34] and fungi [27] [35] [36]. In these two groups, seed endosperm is the feasible and preferable commercial source of gums, as production of seeds can be tailored according to the market demands. There are few species in which galactomannans have been found in non-leguminous seeds, e.g., Coffea arabica, Cocos nucifera, Phoenix dactylifera, Elaies guinensis, Phytelephas macrocarpa, and others. In a few exceptional cases, namely in Glycine max, Gymnocladus diocia, and Mucuna, gum occurs in the hull, the inner side of the seed coat, and kernel, respectively. 3. Natural Function. Endosperm, the reserve source of plant polysaccharides, has dual physiological functions. On the one hand, it serves as a food reserve for germinating seeds, on the other hand, it retains H2O by solvation and, thereby, prevents complete drying of seeds to avoid protein denaturation, in particular the denaturation of enzymes essential for seed germination. 4. Examples of Commercially Used Seed Gums. The most-important commercial sources for seed gums are collected in Table 2. Guar, obtained from the seeds of Cyamopsis tetragonaloba (l.) Taub, is currently the foremost seed gum used worldwide in different industries. Dhaincha and cassia gums, also from Indian sources, are produced in handsome amounts and used in various industries, especially for paper and textile manufacture. Carob and tara gum have Mediterranean sources, and their production is confined to a particular region. The growing industrial utility of these gums at an international level has resulted in an impetus for India to intensify its research and development with respect to new sources of seed gums for commercial production and utilization. 5. Historical Background. The locust bean or carob tree (Ceratonia siliqua l.) has been known to be a pioneer source of seed galactomannans. It originally belonged to Southern Europe and the Near East, and was only later introduced to the U.S.A., Australia, and other parts of the world. Since early times, its pods have been used as human and animal food stuff, and, afterwards, its seeds were used for the isolation of gum to be used in the paper, textile, food, and pharmaceutics industries. Before World

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Table 1. Galactomannans from Leguminosae. The term Man/Gal refers to the experimentally derived ratio of d-manno- to d-galactopyranosyl residues. Species Cassia absus Cassia alata Cassia didymobotrya Cassia juncea Cassia saltiana Leucaena leucocephala Sesbania bispinosa Sesbania grandiflora Sesbania sesban Cassia marginata Crotalaria incana Crotalaria lanceolata Crotalaria retusa Crotalaria spectabilis Medicago hispida Parkinsonia aculeata Trifolium hirtum Cassia fistula Centrosema pulmieri Cassia grandis Cassia laviegata Cassia leptocarpa Caesalpinia cacalaco Cassia spinosa Ceratonia siliqua Delomix regia (flame tree) Gymnocladus dioica Cassia intermedia Cyamopsis tetragonoloba Cyamopsis tetragonoloba Cassia nodosa Cassia occidentalis Cassia sophera Cassia tora Cassia pulcherrima Cassia verrucosa Cyamopsis tetragonoloba Cyamopsis tetragonoloba Cyamopsis tetragonoloba Gleditsia triacanthos Indigofera tinctoria Leucaena glauca Melilotus albus Trigonella cretica Trifolium dubium Trifolium incarnatum Trifolium foenum graecum (fenugreek) Trifolium foenum graecum Trifolium foenum graecum Trifolium hamosa Trifolium monspeliaca Trifolium polycerata Mellilotus officinalis Parkinsonia aculeata Family Caesalpiniaceae Caesalpinaceae Caesalpinaceae Papilionaceae Papilionaceae Minnosaceae Papilionaceae Papilionaceae Papilionaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Papilionaceae Caesalpinaceae Papilionaceae Caesalpinaceae Papilionaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Papilionaceae Papilionaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Caesalpinaceae Paplionaceae Papilionaceae Papilionaceae Papilionaceae Caesalpinaceae Papilionaceae Mimosaceae Papilionaceae Papilionaceae Papilionaceae Papilionaceae Papilionaceae Papilionaceae Papilionaceae Papilionaceae Papilionaceae Papilionaceae Papilionaceae Caesalpinaceae Man/Gal 3.0 3.3 0.5 1.5 3.2 2.3 2 1.8 1.0 2.7 2.7 2.57 2.85 2.85 1.22 2.70 1.04 3.0 0.09 0.7 0.5 3.05 2.5 2.70 3.75 4.28 2.71 2.29 1.54 1.70 2.7 3.5 3.01 1.0 3.0 3.0 4.0 1.77 1.37 2.0 3.2 2.1 1.33 1.13 1.56 1.67 1.17 1.17 1.13 1.2 1.1 1.17 1.08 1.13 1.19 1.6 Literature [4 6] [7] [8] [8] [8] [8] [8] [8] [8] [9] [9] [9] [9] [9] [9] [9] [9] [10] [11] [11] [12] [13] [14] [14] [14] [14] [14] [14] [14] [15] [14] [9] [15] [16] [17] [18] [19] [20] [21] [11] [22] [23] [24] [25] [26] [27] [10] [11] [28] [29] [29] [29] [29] [29] [32] [33] [29] [29] [29] [30] [31]

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CHEMISTRY & BIODIVERSITY Vol. 2 (2005) Table 2. Typical Sources of Seed-Gum Galactomannans

Common Name Guar Carob or locust bean Tara Dhaincha Cassia

Source Cyamopsis tetragonoloba Taub. Ceratonia siliqua l. Caesalpinia spinosa Linn. Sesbania bispinosa Jack. Cassia tora Linn.

Habitat Annual Tree Tree Annual Annual

War II, seed-gum production from carob was basically the monopoly of some western countries. But, during the war, gum production was reduced to a great extent due to difficulties in seed supply from Mediterranean countries. After the war, guar gum from the seeds of Cyamopsis tetragonoloba (L.) Taub. emerged as a new and rival source of carob gum. These seeds contain ca. 42% of gum (as compared to 48% in carob), and in both seeds, the source of gum is endosperm. Guar is an annual shrub, 1 3 m high, and does not require much care in cultivation. Contrary to guar, carob tree takes a long time to mature, and it fruits ripen only after five years. Both species can be grown on poor soil, but collection of seeds is comparatively laborious for carob tree, as it grows up to heights of ca. 12 m. Owing to these factors, guar gum has become a commercial commodity in the international market. Guar legume has been grown in India for a very long time, both for green-manuring purpose and for animal foods. Its pods have also been used as a source of vegetables. The hardy and drought-resistant plant can be grown on sandy loam and deep alluvial soil. In India, there are several varieties of guar [37] that differ from each other in plant height, shape, and size of the pods. The two most commonly recognised varieties are giant and dwarf. The former is grown in Gujarat, while the latter is more popular in Punjab and Uttar Pradesh. In Maharashtra, three varieties are grown: 1) the so-called Desi Guar, a plant ca. 1.5 m high, grown as a dry crop mainly for seeds; 2) Pardesi Guar, ca. 1.8 m high, grown mainly for pods to be used as a vegetable; and 3) Sotia guar, 2.4 3 m high, used to provide shade or for green-manuring purposes. On average, some 450 500 kg of seeds can be harvested per acre. Moreover, ca. 5000 6000 kg of fodder for life stock are obtained per acre. 6. New Sources of Seed Gums. Although much work has been performed on different seed galactomannans, particularly on their composition and structure, very little attention has been paid to comprehensive chemical screening. The U.S. Department of Agriculture (USDA) has carried out commendable work on survey and screening of mucilaginous seeds. Tookey et al. [9] have examined 175 species of 26 plant families as potential sources of water-soluble seed gums. Of 88 species of 32 leguminous genera, some 31 species had gum contents below 10%, 29 species contained 10 15%, 17 species 15 20%, and 11 species more than 20%. Non-leguminous families, e.g., Compositae (twelve species), Malvaceae (eight species), and Umbelliferae (seven species) afforded less than 10% gum, and Convolvulaceae (four species) had 10 15% gum. In total, 21 leguminous species were analysed for their Gal/Man ratios, which ranged from 1 : 1 to 1 : 4.

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In another communication from USDA, Tookey and Jones [38] screened 300 species of 31 plant families for water-soluble gums, out of which 163 leguminous species were found to possess gum contents ranging from 10 to 18%. It was also observed that, in legume seeds, gums were present mostly in the endosperm. Earlier, Anderson [14] had investigated seeds from 163 species of legumes, 75% of which were found to contain mucilage-yielding endosperms. In the search for water-soluble gums at the National Botanical Research Institute (NBRI; Lucknow, India), a chemical screening of more than 300 Indian species [4, 39 46] has been conducted. Thereby, on the basis of previous screening experiments, it was considered desirable to lay emphasis on Leguminosae. The seeds were collected from different parts of India and botanically identified at the seed herbarium of NBRI. The seeds were investigated for water-soluble-gum and moisture contents, and some isolated gums were also analysed for their protein and pentosan contents. In the above studies, several useful observations were made. Seeds belonging to Indian species of Cassia, Crotalaria, Indigofera, Melilotus, and Sesbania were found to be very rich in gum content, whereas all the species of Acacia, Albizzia, Erythrina, Canavalia, Tephrosea, and Millettia were found to be devoid of gum. A few species of genera like Bauhinia, Caesalpinia, and Desmodium contained appreciable amounts of gum. Regarding the richness for both galactomannan and protein contents in seed material, it may be stated here that the association of edible protein with galactomannan helps in the ultimate economic use of the whole material. The gum is invariably present in the endosperms of the seed and is, therefore, easily separable by mechanical means. The rest of the material, i.e., the seed meal, is commonly used as cattle and poultry feed. The seeds of the genus Crotalaria have been found to be rich both in gum and protein, in contrast to those of Cassia. The latter, however, are very rich in galactomannan content (10 39%) and have easily separable endosperm, but low protein content. Some Crotalaria seeds are generally found to be toxic to animals and, therefore, the meal obtained after mechanical separation of the gum is not suitable as feed. This problem, fortunately, can be circumvented by roasting the seed meal, which degrades certain toxic amino acids like mimosine. These studies have also revealed that the subfamily Papilionaceae is generally represented by high amounts of gum as well as protein, whereas high amounts of gum but low amounts of protein, have been generally found for seeds of Mimosaceae and Caesalpinaceae. For commercial gum production, annual and perennial shrubs or herbs are considered more preferable than tree legumes. Ease in mechanical separation of endosperm from the seeds, which mostly depends on the shape and size of the seeds, facilitates the commercial exploitation of new sources. It is to be pointed out here that the seeds of Indigofera, Melilotus, and Desmodium species, though rich in gum content, are very small, weighing less than 10 mg/seed. Likewise, many seeds from Crotolaria species are small, e.g., C. pumila, C. albida, C. calcycina, C. mysorenses, and C. prostrata. Although these seeds are rich in gum content, separation of the endosperm is not feasible. Cassia seeds are generally medium-sized or even bigger; they contain more than 40% of endosperm and are, therefore, considered preferable for commercial exploitation. Some of the possible sources of seed gums have been identified: Cassia

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alata, C. angustifolia, C. didymobotrya, C. grandis, C. nodosa, C. occidentalis, C. saemaea, C. hirsuta, and C. spectabilis. It is to be mentioned here that Cassia angustifolia is a shrub that is being cultivated on large scale in southern India for its leaves and pods. Its seeds are medium-sized, and contain ca. 50% of endosperm. These seeds could, thus, open up a new avenue for the production of seed gum. 7. Chemical Structures of Seed Galactomannans. Seed galactomannans, generally known as seed gums, are heterogeneous polysaccharides composed of galactopyranosyl (Gal) and mannopyranosyl (Man) residues. Generally, classical galactomannans possess a main chain of b-(1 ! 4)-linked Man units, and Gal units are attached to this main chain via a-(1 ! 6)-glycosidic linkages (for exceptions, see Sect. 7.3) 7.1. Weakly Branched Classical Galactomannans. 7.1.1. Guar (Cyamopsis tetragonoloba). The pioneering work on the structure of mannopyrans from plant seeds was mainly conducted with the commercially important guar. Guar contains a b-d-(1 ! 4)linked Man backbone to which are attached single a-d-Gal stubs in 6-O-position of selected d-Man residues. Guar gum contains approximately twice as many a-d-Gal stubs compared to gum from locust bean, as determined by methylation analysis, periodate oxidation, and partial-hydrolysis experiments. Methylation yields mixtures of 2,3,4,6-tetra-O-methyl-d-galactose, 2,3,6-tri-O-methyl-d-mannose, and 2,3-di-O-methyl-d-mannose, whereas partial hydrolysis yields 6-O-a-d-galactopyranosyl-d-mannose and 4-O-b-d-mannopyranosyl-d-mannose. The data from such methylation experiments suggest that the repeating unit of guar galactomannan is composed of three hexose units in which two Man residues are present in the main chain, Gal forming the side chain. The three fractions were obtained in equimolar proportions, as shown by Ahmed and Whistler [47], and Rafique and Smith [48]. The structure of guar galactomannan can, thus, be represented by formula A.

7.1.2. Carob Bean and Gleditsia amorphoides. The galactomannan isolated from Gleditsia amorphoides seed [49] contains d-Man and d-Gal in a ratio of 2.7 : 1. From

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methylation and periodate experiments, structure B1 can be proposed, in which the main chain consists of (1 ! 4)-b-d-Man units, one out of three Man residues carrying an a-d-Gal residue in 6-position. The general structural features of this galactomannan are very similar to those of carob-bean gum [50] [51], for which formula B2 is valid. There are numerous other weakly branched seed galactomannans. Most of them share the basic structure C, but vary in Man/Gal ratio. A lists of such compounds is given in Table 3.
Table 3. List of Weakly Branched Seed Galactomannans. The term Man/Gal refers to the experimentally derived ratio of d-manno- to d-galactopyranosyl residues.

Source Cyamopsis tetragaonoloba (Guar) Ceratonia siliqua (Carob) Sesbania bispinosa Gleditsia amorphoides Gleditsia triachanthos Cassia nodosa Cassia occidentalis Cassia fistula Cassia grandis Cassia alata Cassia didymobotrya Cassia siamea Cassia spectabilis Cassia ovata Cassia renigera Crotolaria verrucossa Delomix regia

Man/Gal 2.0 4.0 1.9 2.7 0.38 0.67 3.5 3.0 3.0 3.2 3.3 3.0 3.07 2.65 2.5 2.6 4.0 2.0

Repeating unit x 1 1 10 10 10 2 1 1 10 10 1 1 10 2 10 2 1 y 1 3 9 12 26 5 2 2 27 33 2 2 16 3 6 6 1

7.2. Highly Branched Classical Galactomannans. 7.2.1. Fenugreek Galactomannans and Related Compounds. The galactomannan isolated from fenugreek seeds contains Gal and Man in a ratio of 5 : 6 [18]. The polysaccharide was completely methylated and hydrolysed. Fission of the products gave mainly tetramethyl and dimethyl hexoses, with only a small amount of trimethyl hexose being formed. The sugars were identified as 2,3,4,6-tetra-O-methyl-d-galactose, 2,3,6-tri-O-methyl-d-mannose, and 2,3-di-O-methyl-d-mannose in a proportion of ca. 5 : 5 : 1, respectively. In periodate oxidation, 5 equiv. of formic acid (HCOOH) were formed for every eleven hexose units, with the consumption of 16.4 equiv. of periodate. The yield of HCOOH corresponded to 45.6% of hexose units, occupying branching positions [52]. Partial hydrolysis of the galactomannan afforded three oligosaccharides identified as 4-O-b-d-mannopyranosyl-d-mannose, 6-O-a-d-galactopyranosyl-d-mannose, and O-b-d-mannopyranosyl-

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(1 ! 4)-O-b-d-mannopyranosyl-(1 ! 4)-d-mannose. These data are consistent with a polymer structure involving a main chain made of b-d-(1 ! 4) Man units, with a-d-Gal residues attached at various intervals to the O-atoms of C(6) of Man. Fenugreek galactomannan can be best represented by structure D, where x z 3, and y 1 (Table 4). In this type of galactomannan, 85% of the main chain is branched.
Table 4. List of Highly Branched Seed Galactomannans. The term Man/Gal refers to the experimentally derived ratio of d-manno- to d-galactopyranosyl residues.

Source Fenugreek Medicago lupulina Lotus pendunulatus Clover Lucerne Lotus corniculatus Green palmyra palmnut Anthyllis vulneraria White clover Soybean hulls Sesbania grandulifera Melilotus officinalis

Man/Gal 1.20 1.10 1.16 1.14 1.25 1.25 1.25 1.33 1.53 1.50 1.67 1.09

xz 3 8 4 5 2 2 2 1 1 0 1 9

y 1 1 1 1 1 1 1 1 1 1 2 1

Galactomannans isolated from the seeds of lucerne [53], clover [54], soybean hulls [54], Lotus pendunulatus [55], Lotus corniculatus [56], white clover [57], Anthyllis vulneraria [58], and Medicago lupulina [59] have been structurally examined by different workers. The first six galactomannans, on methylation followed by subsequent hydrolysis, yielded 2,3,4,6-tetra-O-methyl-d-galactose, 2,3,6-tri-O-methyl-d-mannose, and 2,3-di-O-methyl-d-mannose in various amounts. Periodate-oxidation results were in agreement with the methylation data, but the structures of the last two galactomannans were determined on the basis of identification of oligosaccharides obtained during partial hydrolysis and periodate oxidation. Partial hydrolysis of these galactomannans resulted in the isolation of four oligosaccharides. Fractions I and II were identified as 4-O-b-d-mannopyranosyl-d-mannose and 6-O-a-d-galactopyranosyl-d-mannose, respectively. Fraction III was identified as a mixture of two trisaccharides: O-a-d-galactopyranosyl-(1 ! 6)-O-b-d-mannopyranosyl-(1 ! 4)-d-mannose, and O-b-d-mannopyranosyl-(1 ! 4)-O-[a-d-galactopyranosyl-(1 ! 6)]-d-mannose. Fraction IV was an unidentified mixture of three tetrasaccharides, representing one Gal unit (from side chains) along with three Man units (from the main chain). Similar to fenugreek, there are other highly branched galactomannans, typically containing 65 91% of branched units in the main chain. Medicago lupulina and Lotus

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pendunulatus, for example, contain 91 and 86% branching units in the main chains, respectively, whereas the galactomannan from soybean hulls contains only 63% branching units. The galactomannans isolated from lucerne contains ca. 80% branching units in the main chain, and the seed galactomannans from both white clover and Anthyllis vulneraria contain 75% branching. In the case of clover seeds, of nine hexose units of the main chain, seven are substituted, corresponding to 78% branching. 7.2.2. Galactomannans from Green Palmyra Palm Nut (Borassus flabellifer Linn.). Apart from leguminous seeds, there are some other seeds with mannans that show highly branched structures of type E, as, e.g., in green palmyra palm nut.

In spite of having the same basic structure, the gums obtained from particular species differ from each other due to variations in the Gal/Man ratio as well as in fine structure regarding the attachment of single Gal units to the Man backbone. These factors are mainly responsible for the variation in gum characteristics. The Gal/Man ratio in E often strongly varies for the different species. The mostprominent source of seed gum, i.e., guar, contains a Man/Gal ratio of 1.8 : 1. This ratio is almost identical in the commercial Dhaincha gum. The gums obtained from Cassia species generally contain higher amounts of Man, ranging from Man/Gal 2.5 : 1 to 4 : 1. Carob gum from Ceratonia siliqua l., a foremost species of the Mediterranean region, contains an even higher amount of mannose (Man/Gal 4 : 5). A compilation of various sugar compositions for different leguminous species is presented in Table 1. 7.3. Nonclassical Galactomannan Structures. 7.3.1. General Considerations. As mentioned above, in classical seed galactomannans, b-(1 ! 4)-linked d-Man units form the main chain, and single Gal residues occupy side-chain positions via a-(1 ! 6) linkages. Generally, leguminous galactomannans all possess this typical structure, varying simply in branching pattern and degree of d-Gal substitution to the b-d-Man backbone. Galactomannans reported to share this basic structural element include those from the seeds of C. occidentalis [18], C. fistula [24], Gleditsia triaconthos [26], Arenga saccharifera [30], Borassus flabellifer [56], Gleditsia amorphoides [60], Anthyllis vulnerary [61], Trigonella foenum graecum [62], Medicago lupulina [63], Desmoduim pulchellum [64], Lotus corniculatus [65], Lotus pedunculatus [66], Convolvulus tricolor, Cocos nucifera, Annona muricata [67], Ipomoea muricata [68], Cassia nodosa [69], and C. tora [70], among others. These galactomannans mostly yield three sugars during methylation: 2,3,4,6-tetra-O-methyl-d-galactose, 2,3,6-tri-O-methyl-d-mannose, and 2,3-di-O-methyl-d-mannose, but in varying proportions (different Gal/Man ratios). The presence of 2,3,4,6-tetra-O-methyl-d-galactose indicates branched chains, and, in turn, confirms the presence of Gal moieties at branched positions. The relative amount of liberated 2,3,6-tri-O-methyl-d-mannose provides the

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content of unbranched mannose units in the polysaccharide. The isolation of 2,3-di-Omethyl-d-mannose, finally, reveals that these sugars originate from the main-chain Man units, which are branched with Gal, and its proportion shows the extent of branching. Periodate oxidation and Smith type degradations are other tools to confirm certain structural concepts identified in methylation studies. In periodate oxidation, adjacent OH groups are oxidised, giving rise to oxo polysaccharides. Periodate uptake with concomitant liberation of HCOOH can be followed until constant values are obtained. In Smith degradations, the periodate-oxidised polysaccharide is reduced and hydrolysed, which leads to the isolation of polyols. The galactomannans generally yield glycerol ( propan-1,2,3-triol) and erythritol ( butan-1,2,3,4-tetrol), the former from Man units, the latter from Gal residues. The ratio of erythritol and glycerol further confirms the Gal/Man ratio and their inter-linkages. 7.3.2. Examples of Nonclassical Structures.Few galactomannans have been reported to deviate from the above basic structure, mainly differing in the main chain and the branching arrangement. Unrau and Choy [21] [71] reported such structural variation in Crotolaria mucronata and Caesalpinia pulcherrima. Methylation and hydrolysis of the galactomannan (Man/Gal 3 : 1) yielded 2,3,4,6-tetra-O-methyl-d-mannose, 2,3,4,6tetra-O-methyl-d-galactose, 2,3,6-tri-O-methyl-d-mannose, 2,4,6-tri-O-methyl-d-mannose, 3,4,6-tri-O-methyl-d-mannose, and 2,3-di-O-methyl-d-mannose in a ratio of 1 : 23 : 28 : 11 : 4 : 26. The presence of (1 ! 3)-linkages in the d-mannan main chain was further supported by the finding that one in every six Man residues was resistant to periodate oxidation. Smith degradation yielded equimolar amounts of 2-O-b-dmannopyranosyl-d-erythritol and O-b-d-mannopyranosyl-(1 ! 3)-2-O-b-d-mannopyranosyl-d-erythritol, indicating that there are significant amounts of isolated and consecutive b-d-(1 ! 3)-linkages in the backbone. The methylation results also indicated that ca. 6% of the Man backbone residues are (1 ! 2)-linked. Single d-Gal stubs are linked to the main chain by (1 ! 6)-bonds. In similar experiments, Unrau and Choy [21] found that the structure of galactomannan isolated from Caesalpinia pulcherima also differs from the classical type, but to a much lesser extent, 4% of the b-d-Man units in the main being (1 ! 3)linked, and ca. 2% being (1 ! 2)-linked. Smith degradation again indicated that isolated (n 1) and consecutive (n  2) (1 ! 3)-linkages are present in the main chain, i.e., elements of the type b-d-Man-(1 ! 4)-{b-d-Man-(1 ! 3)}n-b-d-Man-(1 ! 4)-b-dMan. The galactomannan of Kentuky coffee bean contains 20% Gal and 80% Man. Generally, galactomannans contain the side chain of single Gal units, but in this case, both Gal and Man units are present as side chains. The ratio of Gal to Man end groups was found to be 6%. The proposed structure, based on methylation and periodate oxidation, corresponds to formula F.

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Kapoor and Mukherjee [4] reported the structure G for the galactomannan from Cassia absus [72] seeds, which differs from the fundamental structure of the galactomannans not only in the mannan backbone, but also in the attachment of the Gal side chains. The galactomannan (Man/Gal 3 : 0), on methylation, yielded 2,3,4,6tetra-O-methyl-d-galactose (2 equiv.), 2,3,6-tri-O-methyl-d-mannose (4.2 equiv.), 2,3di-O-methyl-d-mannose (1 equiv.), and 4,6-di-O-methyl-d-mannose (1 equiv.). Partial acid hydrolysis yielded the disaccharides 2-O-a-d-galactopyranosyl-d-mannose, 6-O-ad-galactopyranosyl-d-mannose, and 4-O-b-d-mannopyranosyl-d-mannose, as well as the trisaccharides 4-O-(6-O-a-d-galactopyranosyl-b-d-mannopyranosyl)-b-d-mannose and 4-O-(4-O-b-d-mannopyranosyl-b-d-mannopyranosyl)-b-d-mannose.

This was the first report of a 2-O-a-d-galactopyranosyl-d-mannose fragment of galactomannan seeds, although this compound had previously been isolated by degradation of the yeast galactomannan from Trichosporon fermentans [45]. From these results, it was deduced that 85% of the main-chain Man residues are b-(1 ! 4)linked, whereas the remaining 15% are (1 ! 3)-linked. Thereby, all of the (1 ! 3)linked Man residues are substituted at the O-atom attached to C(2) by an a-d-Gal group, accounting for ca. 50% of the Gal side-chain stubs. The rest of the Gal stubs are attached to b-(1 ! 4)-linked Man residues via a-(1 ! 6)-linkages, as shown in formula H.

In some seed galactomannans, small amounts of Gal have also been found in the Man main chain, e.g., in Gleditsia ferox [21] [50] [60] [73 77], G. amorphodies [60], and Trifolium repens [73]. Methylation of the anticipated compound I afforded 2,3,4-tri-Omethyl-d-galactose (from Gal), in small proportions relative to the traditional three methylated sugars (see above). In Cassia grandis seeds, Bose and Srivastava [78] [79] reported the presence of Gal along with Man units joined by (1 ! 4)-linkages in the main chain, together with side chains of Gal and Man units attached via (1 ! 3)-linkages (structure J). Upon methylation, 2,3,6-tri-O-methyl-d-galactose, 2,6 di-O-methyl-d-galactose, and 2,3,4,6tetra-O-methyl-d-mannose were found, along with the three traditional methylated sugars. Thereby, the side chains were shown to comprise 33% of Gal and 67% of Man attached via (1 ! 3)-linkages. Gupta and co-workers [80] isolated a polysaccharide from Cassia corymbosa seeds, which, on methylation, gave rise to an extra sugar, 2,3,4-tri-O-methyl-galactose.

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According to the authors, the side chains are composed of two consequent Gal units linked through a (1 ! 6)-bond, and are attached to the main chain by a (1 ! 6)-linkage, as shown in structure K. The same authors reported that the side chain of the seed galactomannan L of Cassia laviegata also contains two consequent Gal units. Thereby, the galactomannan has the untypical Gal/Man ratio of 2 : 1 1).

A water-soluble galactomannan from the seeds of Indigofera tinctoria also deviates from the fundamental structure. Five methylated sugars, namely 2,3,4,6 tetra-Omethyl-d-mannose, 2,3,4,6 tetra-O-methyl-d-galactose, 2,3,6 tri-O-methyl-d-mannose, 2,3,4-tri-O-methyl-d-mannose, and 2,3-di-O-methyl-d-mannose were isolated upon methylation and hydrolysis, suggesting the presence of Gal and Man units in the main and side chain, respectively [81].
1)

The Man content of galactomannans is generally higher than that of Gal.

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Gupta and Bose isolated a galactomannan with a Gal/Man ratio of 1 : 1.14 from the seeds of Mellilotus indica All. (syn. M. parviflora Des.) [82]. The authors have reported a highly branched unique galactomannan with a (1 ! 2)- as well as (1 ! 4)linked Man main chain, and side chains of single and double Gal units, together with four-residue side chains composed of one Gal residue and three Man units attached via (1 ! 6)- and (1 ! 4)-linkages. Accordingly, the following methylated sugars were obtained upon derivatisation: 2,3,4,6-tetra-O-methyl-d-mannose (1 equiv.), 2,3,6-triO-methyl-d-mannose (2 equiv.), 2,3-di-O-methyl-d-mannose (22 equiv.), 3,4-di-Omethyl-d-mannose (6 equiv.), 2,3,4,6-tetra-O-methyl-d-galactose (27 equiv.), and 2,3,6-tri-O-methyl-d-galactose (3 equiv.), indicating structure M.

The seed galactomannans of Parkinsonia aculeata Linn. [83] (Man/Gal 1.85 : 1) has also been found to contain Gal and Man moieties with (1 ! 3)-, (1 ! 4), and (1 ! 6)linkages, main-chain Man and Gal being connected via (1 ! 3)-linkages. The Gal/Man proportion within the main chain was found to be 1 : 2, whereas that of the side chains was 1 : 4, indicating the highly branched character of these polymers. 7.4. Other Structural Aspects of Seed Galactomannans. In most high-molecularweight polysaccharides, the Man residues occupy b-(1 ! 4)-linked main-chain positions, side chains being attached via a-(1 ! 6)-Gal units. These gums, although sharing similar basic structures, may have very different fine structures regarding the distribution pattern of side chains. Generally, three types of patterns are observed: symmetric, random, and block patterns (together with the few variations discussed above). It should be noted that heterogeneous high-molecular-weight polysaccharides are very difficult to characterize, and several techniques have to be adopted to determine at least the most-important structural aspects. Typical experiments include, from a chemical point of view, methylation, periodate oxidation, and degradation, followed by identification of the resulting oligosaccharides. However, the fine structure of galactomannans can be resolved only through NMR spectroscopy, x-ray diffraction, and enzymatic studies. 8. Isolation, Purification, and Physicochemical Characterization of Galactomannans. 8.1. Isolation and Purification. In leguminous plants, seeds are dicotyledonous, and endosperm is the desirable part for the extraction of gum. On a commercial scale,

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seeds are first roasted in furnace to loosen the seed coat from the endosperm, followed by breaking the seed in specified mills by means of different mesh-size sieves. The separated endosperm generally sticks with hull and, so, has to be resubjected to the milling process. Then, the separated endosperm is powdered by means of pulverisers to desirable mesh size. Thereby, special care has to be taken to avoid degradation of the high-molecular-weight polymers. Generally, pulverisation is performed under humid conditions using a spray of steam showers. Pulverisation is a specific technique that can provide a gum powder of lower or higher viscosity, depending on the conditions applied (humidity, temperature, etc.) and sophistication of the infrastructure. The gum so obtained is termed crude gum or commercial-grade gum, and is applied as such in different industries. Crude gum obtained by dry milling generally contains pentosan, proteins, and fibres (etc.) as impurities. Hence, it requires purification before any physico-chemical properties can be determined. For this purpose, the gum is dissolved in H2O, and regenerated by fractional precipitation with polar solvents. Gums can also be purified by complexation with Cu2 and Ba2 salts, or, for advanced studies, they may be further purified by dialysis and membrane filtration. The level of purification can be checked by the gums homogeneity. The constituent-sugar ratio, average molecular weight, intrinsic viscosity, and polydispersity are the important physico-chemical parameters that define the nature of a gum. Molecular weight and sugar ratio may vary considerably, depending on the source and isolation procedure. The Man/Gal ratio typically ranges from 1.1 to 5.0 (see Table 1). 8.2. Solubility and Viscosity. Structural variations, i.e., variations in the Gal/Man ratio and type and degree of substitution, cause significant changes in the solubility, viscosity, and ability of a gum to interact with other carbohydrates. For example, guar gum, which has a higher Gal content (Man/Gal 1.8 : 1) than carob gum (4 : 1) from Ceratonia siliqua (l.), disperses more rapidly in H2O. Generally, polysaccharides with higher Gal contents are readily soluble in H2O, but have a lesser tendency to form gels. It has been postulated that the high solubility of guar gum is attributed to the presence of more side chains, keeping the main Man chains far enough apart to prevent efficient covalent interactions. In contrast, carob gum has the tendency to interact with gelling polysaccharides. Generally, gums with higher Man content (and, thus, fewer side chains), consist of long blocks of unsubstituted Man units that can either covalently interact with or chemically bind other polysaccharides (cross-linking). In fact, there are some deviations from the above basic structures, where some of the Man units are connected through different linkages. In some galactomannans, side chains of glucose and/or Man with different glycosidic linkages have also been reported. 8.3. Rheological Behaviour. Much of the interest in galactomannans has resulted from their unique property to provide aqueous solutions of very high viscosities [84]. Most rheological investigations have originally been commercially motivated, and, consequently, the commercially important gums have been studied in much more detail than the others [85 87]. In aqueous solution, galactomannans predominantly exist in a random-coil conformation [88], and more-ordered forms may contribute to the overall structure only under certain, favourable conditions that allow self-aggregation and/or interaction with other species.

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In the solid state, the Gal side-chain residues seem to have little effect on the conformation of the Man backbone. However, as mentioned above, the side chains play a crucial role in determining the ease with which galactomannans can be dissolved. Hui and Neukom [89] fractionated the gum of locust bean by means of temperaturedependent differential solubilities. The cold-water-soluble fraction contained a high content of Gal, in contrast to the hot-water-soluble fraction. There may be even an insoluble fraction composed of even fewer Gal residues. Locust bean gum is generally difficult to disperse, presumably due to strong intermolecular interactions between adjacent chains. This gum has been implicated as a promoter of the retro-gradation of amylose; it interacts strongly through regions of the backbone sparsely substituted with Gal groups to cause precipitation. Guar gum, having more side chains, disperses more readily than locust bean gum, as the peripheral side chains hold the main chains far enough apart to prevent effective intermolecular interactions. Locust bean gum has been found to be completely dispersed in H2O only after heating at 808 for 20 30 min. This process, which is accompanied by a dramatic increase in viscosity, can be sped up by addition of 6m urea solution at 208: presumably, urea assists in destroying the noncovalent bonds between adjacent chains [90]. Properties such as concentration, shear rate, time, temperature, pH, and other factors greatly affect the rheological behaviour of galactomannans in aqueous solutions [91] [92]. These natural gums, when completely dispersed in H2O at 1% concentration, give rise to highly viscous, non-Newtonian, pseudoplastic solutions. At lower concentration (0.5%), pseudoplasticity is less evident, and the solutions tend to adopt Newtonian properties [93]. The maximum viscosity and the stability of a viscous solution on storage depend very much on the way in which the gum gets dispersed, dispersion time and temperature being particularly important. According to Carlson et al. [94], heating for long periods at temperatures above 608 tends to provide a high initial viscosity, but leads to more-rapid breakdown and inferior stability. Optimal conditions for achieving maximum viscosity of solutions of guar gum involve heating at 25 408 for 2 h, in contrast to locust bean gum, which cannot be fully dispersed at room temperature, the resulting solution being of low viscosity. Other than these gums, various other gums require varying conditions and ways of formation to attain optimal viscosities. For a given range of galactomannans with identical Man/Gal ratios, the viscosities are proportional to the molecular weights of the polymers. However, there is some evidence that gums having a higher content of Gal give rise to higher viscosities. Hui [95] found that a sample of locust bean gum with a molecular weight (Mr ) of 650,000 had a lower viscosity than a sample of guar gum of Mr 250,000. However, this may also be due to presence of aggregates in the sample of locust bean gum, and so, the extent of the Gal-free mannan backbone may be more important in this regard than the actual Man/Gal ratio. Since galactomannan solutions are basically neutral, pH changes not too dramatic have only a moderate effect. The viscosity tends to remain constant over a pH range as broad as pH 1 10.5, although degradation under highly acidic and alkaline conditions occurs, especially above pH 11 and at elevated temperatures, where the rate of dissolution is depressed. Carlson and Ziegenfuss [96] found a procedure for getting

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more-homogenous solutions through elimination of clumping. One possibility is, e.g., to mix the dry gum with sucrose and certain salts of sodium sulfate; the resulting competition for H2O then gives rise to smaller rates of gum dissolution, and, thus, higher homogeneity. The viscosity and thickening properties of galactomannans extracted from the seeds of Leguminosae have found broad applications in food, plant, cosmetics, paper, and pharmaceutics industries [97] [98], one special application being, e.g., petroleumdrilling technology, among others. The galactomannans from the seeds of guar (Cyamopsis tetragonoloba) and carob (Ceratonia siliqua) have found widespread use as industrial high-viscous hydrocolloids. The viscosity of solutions of guar galactomannan has been studied in detail [94] [99 100], and was found to be shear-rate dependent. Apart from this, very little is known about the properties of galactomannans in dilute aqueous solution. Koleske and Kurath [101] have reported a detailed study of solutions of galactomannan acetates. Sharman et al. [84] made a comparison of the hydrodynamic properties of solutions of galactomannans from Leguminosae and guar, and of naturally occurring galactomannans and industrially useful, synthetic, watersoluble cellulose derivatives (hydroxyethyl cellulose and ethylhydroxy cellulose). Robinson et al. [102] discussed the viscosity/molecular-weight relationship, intrinsic chain flexibility, and dynamic solution properties of guar galactomannan. Richardson and Ross-Murphy [103] rationalized the nonlinear viscoelasticity of polysaccharide solutions. Detailed rheological properties of guar and hydroxyethyl guar gum have been studied by Patel et al. [104]. More recently, studies on solution properties of some other seed galactomannans have been made, including Mimosa scabrella [86], Cassia nodosa [105] [106], Cassia pulcherrima, and Cassia javanica [107]. Many other potential sources other than guar and carob are currently being exploited in industry, one example being studies concerning the rheological properties of galactomannans from C. angustifolia seeds, which are readily available in India and, thus, a potential alternative source of commercial gum. 8.4. Galactomannan Polysaccharide Interaction. In industry, galactomannans capable of interacting with polysaccharides have a long tradition [108]. It was before 1816, when Merat catalogued the industrial value of gels in general. Later, Baker and co-workers [109], and Duel and co-workers [110], when investigating the effect of locust bean gum on the mechanical properties of carragennan and agar gels, found that even small additions of locust bean gum make the gels firmer, less brittle, and more elastic. Similarly, measurements of rigidity indicated that locust bean gum, guar gum, and fenugreek seed mucilage interact with agarose, increasing the strength of the gel. Hui [95] has shown that it is mainly the hot-water-soluble fraction of locust bean gum that is responsible for the strengthening of agar gels, which was depleted in d-galactose [25]. The industrial importance of these natural gums is also due to the fact that the raw materials (locust bean, guar) are much less expensive than, e.g., carrageenan and agar. Thus, gels of specified strengths can be made with a mixture made of an inexpensive galactomannan and a small amount of carrageenan or agarose, which reduces the production costs. The resulting mixed gel, although of textural properties different from those of the pure carrageenan and agarose gels, may even be preferable in some applications. Interaction of galactomannans with the exopolysaccharide from the plant pathogen Xanthomonas campestris has gained industrial importance [108] [111] [112].

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Both locust bean gum and xanthan polysaccharide form viscous solutions themselves, but no gels at a concentration of 0.5%. However, a mixture of both compounds, comprising 0.5% of each polysaccharide, gives rise to a firm, rubbery gel with a distinctive texture and mouth feel [113]. Hence, the non-gelling galactomannan possesses the ability of increasing the firmness of agar and carragenan gels, and causes gel formation in the presence of xanthan gum, which indicates their participation in the formation of the gel framework. Non-gelling galactomannans may also specifically interact with different bacterial polysaccharides [88]. Gums are employed frequently for their physical functions such as gelation, viscous behaviour, stabilization of suspensions, emulsions, and foams, or control of crystal growth. Physical manifestations such as gelation and viscosity are interrelated and effect other stabilizing functions that are dependent on specific attributes of flow or gelmicelle arrangement. Carrageenan, agar, pectin, xanthan, etc., either alone or in combination with other hydrocolloids, are used to impart these properties to aqueous systems. Carrageenans, structural polysaccharides from seaweed extract (Irish moss), are polysaccharide sulfate ester salts with a sulfate/hexose ratio close to unity. Carrageenan is a unique, strongly charged anionic polyelectrolyte of large size and regular structure (see below). It is the most strongly charged natural anionic polyelectrolyte known. Complex formation with other molecules, large or small, can take place through a number of mechanisms involving ionic and Van der Walls interactions, which makes this gum especially useful. Carrageenans can be induced to form thermally reversible gels whose strengths and gelling temperatures are dependent on the counter-ions used, particularly K and ammonium ions. A distinctive property of these polysaccharides is their ability to alter, at relatively low concentration, the degree of agglomeration of casein particles in cows milk. In each fundamental chemical repeat of the carrageenans, there is a sulfate. Among the different natural carrageenans, so-called i-carrageenan bears a sulfato group on each sugar residue, k-carrageenan has a sulfato group on each d-galactose, and furcellaran possess a sulfato group on ca. 50% of the d-galactose moieties. Thereby, the sulfate content is a direct measure of the concentration required for gelation: i-carrageenan, e.g., gels at 2.5%, k-carrageenan at 1.5%, and furcellaran at 0.4%, as monitored by measuring optical rotation [114]. Anderson et al. [115], and Arnott and Scott [116] have revealed through detailed X-ray analysis that i-carrageenan molecules consist of parallel, exactly staggered, righthanded double helices, with three disaccharide residues per turn and a pitch of 26 (Na salt) or 26.6 (Ca salt). The secondary and tertiary structures of k-carrageenan and furcellaran were found to be less-well-defined, but are anticipated to consist of parallel, threefold, right-handed double helices slightly displaced from an exactly staggered position, the pitches being 25 . In all of these three ordered conformations, the sulfato groups lie on the outside of the double helices and are, thus, solvated [115] [117]. Despite widespread technological use and interesting questions concerning the synergistic interactions between different hydrocolloids in solution, the molecular origin of such effects has, in general, remained obscure. The interaction of plant galactomannans with agarose or k-carrageenan to form mixed gels, at total polymer

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levels far below those required for gelation of either polysaccharide, has been studied. The interaction involves intimate association of specific sequences of the galactomannan with native double-helical stretches of the algal polysaccharide chains [118]. Recently, a similar mechanism has been proposed for the interaction of galactomannans with xanthan gum, a heteropolysaccharide of bacterial origin. The authors have synthesized xanthan produced by the well-studied bacterium Xanthomonas campestris, investigating the biological role of substrate recognition. However, the results diverged on many aspects such as the nature of the ordered structure [119] or the influence of substituents on solution properties [120]. The first investigation with respect to the viscoelastic properties of xanthan gum had been made in 1981 by Thurston et al. [121]. A few studies have revealed the existence of two different ordered xanthan conformations in solution (native vs. renatured), which give rise to different hydrodynamic and rheological properties, but not affecting the position of the conformational transition [122] [123]. The xanthan conformation in solution has been modelled as a worm-like chain characterized by an intrinsic persistence length of either ca. 500 or 1200 . Ross-Murphy and co-workers [124] [125] found that the solution viscoelasticity is drastically modified when either the counter-ion of the polyelectrolyte is changed or when the solution is treated with urea. However, interestingly, in all cases examined, the locally ordered xanthan structure remained unaffected. Roechefort and Middleman [126] further studied the influence of salt, temperature, and strain effects on xanthan solutions prepared from dried xanthan powder and a pasteurized fermentation broth. More recently, steady and dynamic shear properties of aqueous solutions, including xanthan, have been published. The rheological properties of xanthan depend not only on the nature of counter-ions and ionic strength, but also on post-fermentation like pasteurization and drying [127], which also affects the gel-like behaviour [121] [124 126]. Milas et al. [108] studied in detail the flow and viscoelastic properties of xanthan gum solutions using xanthan from pasteurized broth. Recently, Andrade et al. [128] studied the rheological properties of mixtures of k-carrageenan and galactomannan from C. javanica. 9. Modification and Derivatisation of Galactomannans. Seed gum industry in India is mainly devoted to the production and export of unmodified, higher-grade gums; the present scenario of value addition, thus, being unsatisfactory [129 131]. Technically advanced countries rather prepare modified gums for the production of large-value-added products. It is, thus, not surprising that value-added gum products are even re-imported into counties of origin. If such modified products could be made in India and exported, there would be a manifold increase in foreign-exchange earning from the gum industry. It is to be emphasized here that reprocessing, modification, and derivatisation of seed gums require medium-level technology, which can be acquired easily, but requires active collaboration amongst researchers, technologists, and entrepreneurs. In spite of the tremendous usefulness of gums of different types, so far, little has been done to prepare and examine gum derivatives in developing countries. Locust bean and guar gums react smoothly with 1-chloro-2-cyanoethane ( 3chloropropanenitrile) in alkaline solution to give the corresponding 2-cyanoethyl ethers [132] [133]. These gums also react similarly with 2-chloroethanol and with

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ethylene oxide ( oxirane) to give the corresponding O-(2-hydroxyethyl)-functionalized gums [134 136], which are readily soluble in H2O. Locust bean gum reacts with 2chloro-propan-1-ol in alkaline solution to give elastic, irreversible gels [134 138]. O(Carboxymethyl) derivatives of guar and locust bean gum, made with chloroacetic acid, form viscous aqueous solutions that are stable to alkaline reagents [133] [137 140]. Acetates of guar [141] [142] and of locust bean gum [143], which are insoluble in H2O, might be useful for making films [138] [141]. By a similar series of reactions, the poorly water-soluble laminarin was transformed into the corresponding, much more soluble 2-hydroxyethyl, 2-hydroxypropyl, and carboxymethyl ethers, some of which may be suitable as blood extenders [144]. The solubility of laminarin can also be enhanced by formation of sulfates. Similarly, an industrially important alginic acid derivative, the corresponding 2-hydroxypropyl ester, can be made by the reaction of propylene oxide ( 2-methyloxirane) with swollen alginic acid under mild conditions, which produces little or no degradation. Unlike the parent alginic acid gum, this derivative is considerably less sensitive to the addition of acids and remains in solution. Analogous esters have been made from various epoxides, including long-chain epoxides such as 1,2-epoxydecane or the 9,10-expoxides of both stearic acid and stearyl alcohol [145 148]. All these esters are hydrolyzed under basic conditions. Acetates of alginic acid [149 153] and laminarin [154], though prepared successfully, have found little use in industry. Alginic acid, on reaction with mustard gas, forms a thiodiethylene derivative [154]; it has also been reported to react with double isocyanates [155] [156]. In recent years, attention has been directed to the preparation of synthetic blood anticoagulants as substitutes for heparin. For this purpose, sulfates of guar and locust bean gum [138], agar, laminarin [144] [157], dextran [158 160], starch [161 167], glycogen and cellulose [163 167], polyvinyl alcohol [163] [168], chondroitin sulfuric acid and xylan [169], and synthetic glucans [170] have been investigated, respectively. Some of these derivatives have activity in vivo, but, so far, none has matched the activity of heparin. Certain sulfamic acid derivatives, e.g., the sulfated 2-aminoethyl ether of laminarin, display approximately half the activity of heparin [157]. Paritol (alginic acid sulfate) [171] is not more toxic than heparin, but its effects last twice as long. Also, extracts of Chondrus crispus have ca. 40% of the activity of heparin. Phosphorylated derivatives of polysaccharides can be made in the usual way, but little information is available concerning their properties and possible applications. Perhaps such substances could be used both as soil conditioners and fertilizers [172 174]. Carboxymethyl cellulose (CMC) is used as a binder and thickener. It retards icecrystal growth and aids to stabilise emulsions. As its viscosity drops during healing, it may be used to improve the volume yield during baking by encouraging gas-bubble formation. Its control of viscosity allows CMC to be used as a phase thickener, emulsion stabilizer (e.g., with milk casein), and suspending agent. It can also be used for its high water-holding capacity, which is significant even at high viscosities, particularly when applied as the Ca2 salt. Thus, CMC is used for retarding staling and reducing fat uptake into fried food. In 2000, the value of cellulose ethers on the world market was nearly 2 billion U.S. dollars, and world consumption amounted to ca. 370,000 metric tons.

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Thereby, Western Europe consumes the largest amount of cellulose ethers, followed by the Soviet Union and Japan, CMC being the major cellulose product consumed.
The authors are grateful to the National Botanical Research Institute (NBRI) of India for providing research facilities. The senior author is also thankful to the Ministry of Health and Family Welfare, India, for financial support.

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Received July 26, 2004