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Journal of Theoretical and Experimental Biology (ISSN: 0972-9720), 8 (3 and 4): 147-151, 2012 © 2012 Elias Academic

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Chemical and Spectral Characterization of Cell Wall Polysaccharide of Gracilaria edulis (Gmelin) Silva
B. Madhaiyan and N.Rajasulochana*
Department of Plant Biology and Plant biotechnology, Presidency College, Chennai-600 005, Tamil Nadu, India.

Received: 12 January, 2012; revised received: 26 May, 2012

Abstract
The red alga Gracilaria edulis (Gmelin) Silva (Gigartinales) was collected from Rameswaram coast of Tamil Nadu. Proximate analysis was carried out. After extracting the phycocolloid, the yield and physico-chemical properties were determined. The gel strength of agar was found to be dependent on higher 3,6anhydrogalactose content and inversely related to the sulphate. The cell wall polysaccharides extracted from G. edulis was analysed by FTIR spectroscopic method. Keywords: Gracilaria edulis, cell wall polysaccharide, phycocolloid ,anhydrogalactose.

Introduction
Seaweeds contain large amount of carbohydrates, proteins, minerals, trace elements and vitamins. Gracilaria species are also utilized as human food, mostly in salads and soups and also as feed for marine animals. Having this in mind, proximate analysis of Indian seaweeds were carried out by several algologists in different Coasts (SumitraVijayaraghavan et al., 1980; Chennubhotla et al., 1990; Kaliaperumal et al., 1994;Ganesan and Kannan, 1994; Sarojini and Subbarangaiah, 1999; Roslin,2003). The major value of some seaweed lies in their relatively large content of hydrophilic colloidal polysaccharides such as agar, algin and carrageenan. Agar is of great importance in the food and drug industries and has found several applications in bacteriology, plant tissue culture, biochemistry and molecular biology. Despite its high price, increasing with its degree of purity, agarose cannot be challenged by other colloids (alginates, carrageenans, etc.) in its specific uses (Armisen and Galatas, 1987).The polysaccharide agar is composed of repeated units of 3-linked β-D-galactose and 4-linked α-galactose units. The production of agar by seaweeds shows considerable variations in gel quality and yield from different algal species and strains (Cote and Hanisak, 1986) due to seasonal fluctuations. Agar has many applications depending on its quality or properties. The yield and physico-chemical properties of phycocolloids obtained from various species of agarophyteswere recorded in earlier works (Duckworth et al., 1971; Oza ,1978; Chennubhotla et al., 1986; Kaliaperumal et al.,1990; Sasikumar et al., 1999; Vimalabai et al., 2003). The alkali treated agar showed low sulphate and with the high 3, 6-anhydrogalactose content in Gracilaria sp. (Villanueva et al., 1999; Praiboon et al., 2006). FTIR spectroscopy has been used as an effective tool to identify the type of polysaccharides.The spectral analysis of the agar from Gelidium madagascariense, Gelidiella acerosa and Gracilaria millardetii showed the presence of agar polymers, with an intense peak at 890 cm-1, characteristic of a non sulphatedosidic ring as found in agar (Stancioff and Stanley, 1969; Villanueva et al., 1999; Rajasulochana, 2009). Most of the 4 linked units are found as 3, 6 anhydrogalactose, resulting in an intense peak at 930 cm-1.As the information available is very scanty on the spectral studies of agars in India, the present study is focused to investigate the physico-chemical properties and FTIR spectroscopy of polysaccharide extracted from Gracilaria edulis (Gmelin) Silva.
*Corresponding author; Email address: rajasulochana_n@ymail.com

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2003). The carbohydrate and lipid content recorded in the present study is in conformity with earlier reports (Sumitra Vijayaraghavan et al. 1999). The polysaccharide so obtained was dried. 1980. Values were expressed as percentage of polysaccharide dry wt (% ps dry wt).. It is quiet natural. 1951) and lipids (Folch et al..5% phycocolloid (Hellebust and Craigie.. Roslin . 1999. Gelling ability of phycocolloids depends on the molecular weight and level of 3. proteins (Lowry et al. The percentage yield of the dried polysaccharide was calculated from the dry weight of the original sample (raw material). India. 1957). The gel strength of agar of G.. 1956. The dried algae were powdered and used for estimation of total carbohydrates (Dubois et al.. 1990. 1956). mud and epiphytes and then shade dried.edulis is comparable with earlier findings (Kaliaperumal et al. melting temperature and gel strength were determined for 1. Gelling and melting temperature is dependent on molecular weight distribution. Total galactosewas estimated by the phenol sulphuric acid method (Dubois et al. Sarojini and Subbarangaiah.. 8 (3 and 4): 147-151.edulis showed gel formation at room temperature..The chemical composition of seaweeds varies with species.Madhaiyanand Rajasulochana / Characterization of Cell Wall Polysaccharide of Gracilaria edulis Materials and Methods Thalli of Gracilaria edulis(Gmelin) Silva(Gigartinales) were collected from the coast of Rameswaram. Salinity and temperature affect the gel strength of agar from Gracilaria. (1965). 6 anhydrogalactose (Murano et al. 2003). The carbohydrate content was recorded maximum(21%) than the protein and lipid in the present study (Table1). 2012 148 . The gel strength showed positive significant correlation with melting temperature. habitats.resorcinol method of Yaphe and Arsenault (1965) and sulphate by following method of Verma. Chennubhotla et al.. (1977). since the seaweeds usually accumulate smaller quantities of fatty acids and lipids. 1994. The samples were washed in seawater to remove sand. 1990). Spectrum was recorded using Bruker Vector 20 FT-IR double beam Spectrophotometer in the range of 4000-600 cm-1.. Table 1: Biochemical analysis of Gracilaria edulis. powdered. edulis is 106 g/cm2. The phycocolloid obtained from G. 6 AG) was estimated following the acetal .. Extraction of phycocolloid was done by the method adopted by Visweswara Rao et al. The gel strength of phycocolloid is negatively correlated with sulphate content (Villaneuva et al.Gelling temperature. 3. maturity and environmental conditions.).. Results and Discussion Proximate Analysis The moisture and ash content of Gracilaria edulis were observed as 10 % and 21%. 6 anhydrogalactose (3... 1999).This is comparable with earlier observations (Sasikumar et al. Journal of Theoretical and Experimental Biology (ISSN: 0972-9720). 1986). under identical conditions and compared with spectrum of commercial agar. The higher carbohydrate content might be due to the phycocolloid content of the cell wall (Dhargalkar et al. These are in conformity with earlier report (Chennubhotla et al. TamilNadu. 1980). Parameters Moisture (%) Ash(%) Carbohydrate(%) Protein(%) Lipid(%) 10 21 21 12 2.edulis (26%). The gelling and melting temperature of agar extracted from Gracilaria edulis is 38°C and 80°C.3 Physico-chemical Properties of Phycocolloid The physico-chemical properties of phycocolloid are given in Table 2. 1978). The protein content of G. confirming their agar nature. transferred to air tight glass bottle and stored for quality analysis. Vimalabai et al. The biochemical studies revealed the order of concentration was carbohydrate> protein>lipid. Ganesan and Kannan.. The yield of agar was obtained from G. 1994.

edulis exhibited more or less common constant peaks around 620. 2925. (1971). edulis was performed to determine the composition of total galactose. 2012 149 . spectrum of G.. 1380. 1150. 1640. 1080. 6 anhydrogalactose and sulphateIn the present study. (1999) in species of Gelidium and Gracilaria.Madhaiyanand Rajasulochana / Characterization of Cell Wall Polysaccharide of Gracilaria edulis Table 2: Physico-chemical properties of Gracilaria edulis. 3.2 The chemical analysis of G. 1720. FT-IR Spectral Analysis As in Difco agar. 890.edulis is similar to the findings of Duckworth et al.. Some additional peak exhibited in the study sample (Fig. 930. Figure 1: FTIR Spectrum of polysaccharide from Gracilariaedulis compared with Difco agar. 2850. 8 (3 and 4): 147-151. Sasikumar et al.1). 6 anhydrogalactose and sulphate content contributed to its quality viz. Parameters Yield (%) Gelling temperature( °C) Melting temperature( °C) Gel strength(g/cm²) Galactose(%) 3.6anhydrogalactose(%) Sulphate (%) 26 38 80 106 36 36 2. and 3410 cm-1.6 anhydrogalactose and sulphate content of G . Galactose content of phycocolloid played a significant role in its yield and the 3. 670. The 1740 cm-1 band galactan is assigned Journal of Theoretical and Experimental Biology (ISSN: 0972-9720). 3. (1999 ) and Villanueva et al. 1250. The occurrence of peak at the 2925 cm-1 signifies C-H asymmetric stretching. 1420. the galactose.. gel strength. 1020. 770.

W.K. 1999. Gilles.R. V..M. PanigrahyR. M. Mar. cm-1 are also due to ester sulphate.S.Colorimetric method for determination of sugars and related substances.. N... Ass.G.. F. A. Physiological and Biochemical methods. The linking of carbon-sulphur is determined by the peaks 637 and 708cm-1.. 1994. properties and uses of agar. M. Miller. 512. Kalimuthu S. M. and Bodard. Dhargalkar. D.Seaweed Res.Studies on phycocollloid contents from seaweeds of South TamilNadu Coast.P...S... Seaweed Res. Chennubhotla. Chem. 1990. The peaks of 931and 960 cm-1 very well correspond to the interpretation of Stancioff and Stanley (1969) as due to the vibration of 3. Cote. Biochemical composition of some common seaweeds from Lakshadweep.and Ramalingam.. The peaks at around 1070.1983.S.FAO Technical paper..S. 36:316-319. CMFRI Bulletin.K.A.G. 1986.. 1150. 1980. 1986. Biochemical constituents of seaweeds along Maharashtra coast. 6 anhydrogalactose. and Galatas. R.. 26: 425-427. pp. 9: 297-299.. Less M. J. M. Duckworth. Biol. 856 and 878 cm-1 were exhibited in G. K. 1999). J.J. Mar. RamalingamJ. J.and Kaliaperumal.A. J. The peaks at 817 and 878 cm-1 for ester sulphate in C-6 and peak at 856 cm-1 is ester sulphate in C-4.. N. Res.. 1...G.A simple method for the isolation and purification of lipids from animal tissues. Similar observation has been made by many workers (Christiaen and Bodard 1983..J.Production and properties of native agars from Gracilaria tikvahiae and other red algae. 288: 1-57. Rebers.. 28: 350-356. M. 226: 497-507. 8 (3 and 4): 147-151..R. and Smith.and Kalimuthu.H. V. 1956. 1956. 2012 150 .. Bot..Madhaiyanand Rajasulochana / Characterization of Cell Wall Polysaccharide of Gracilaria edulis to C=O stretching of carboxylic group but this peak is not exhibited in spectrum of G. hence it can only be recommended as a food grade agar. F.Phykos. 1990. V. 2003).Camb.D. 1983. Utiln. Folch. Najmuddin. and Craigie. Indian. The OH / NH stretching mode occurs at around 3400-3500 cm-1 with very strong intensity of bands. This is comparable with previous observations (Villanueva and Montano. Villanueva et al.S. Najmuddin M. Stoare-Stanley. Ramalingam.A. Mar. Najmuddin. Kalimuthu. 1971.. 1987. N.. K. G. Rajasulochana et al.. 16:189-197.. Chennubhotla.. and Yaphe. Journal of Theoretical and Experimental Biology (ISSN: 0972-9720)..K.S.. edulis.Biochemical composition of some macroalgae from Mandapam coast.Bot.Anal. Hamilton. TamilNadu. The additional peaks at 817. 1250. Dubois. Changes in growth and phycocolloid content of Gelidiella acerosa and Gracilaria edulis. The band at 890cm-1 is typical of non sulphated β-D galactopyranose residues of agar.12(1 and 2): 115-119. Seasonal variation in the biochemical constituents of economic seaweeds of the Gulf of Mannar. Kaliaperumal.Press...The structure of agar.and Untawale. This observation is similar to earlier findings (Troung et al.J.. Biochem.. As the gelling temperature of agar from Gracilaria edulis is high (38°C) unlike the general gelling temperature of the bacteriological agar which ranges from 32-36°C. and Selvaraj. Ganesan.. Mar. This result is similar to earlier observations (Christiaen and Bodard. Jagatap. 1994. India.L.K. 9: 45-48.. Christiaen.Carbohyd. Kaliaperumal.In: (ed. Production. This may be due to generic and species variation. Further research towards addition of certain chemicals to bring down the gelling temperature may help in making the bacteriological grade of agar. Strong and very sharp peak at 1640 cm-1 is observed in the present study for amide I deformation / C=O asymmetric stretching.R. and. Utiln. In the present study the peaks at around 1450 cm-1 and 1370 cm-1 noticed in both the spectra of polysaccharides. J. Hellebust. 1988). and Hanisak.) Production and Utilization of products from commercial seaweeds.V. References Armisen. Sci.Spectroscopie infrarouge de films d’agar de Gracilaria verrucosa( Huds) Papenfuss.J.. 44: 442-446.Univ. M. 29: 359-366.This may be due to developmental variation associated with enzymes involved in the synthesis of polysaccharides.33 (1 and 2): 125-135. Chennubhotla.L. Cambridge. 2009). Hand book of phycological methods. 1978. Edited by: McHughs.J. and Kannan.Fractionation of a complex mixture of polysaccharides. M. which are attributed to asymmetrical and symmetrical stretching of CH3/CH2 groups respectively. edulis.

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