EXTRACTION OF GALACTOMANNANS FROM GUAR BEA|NS AND THEIR APPLICATION AS EDIBLE COATING ON CHEDDAR CHEESE

ABSTRACT Cheese is a fermented food which is quite unstable in nature. Cheddar cheese is a hard cheese with firm body having close texture and nutty flavor. Galactomannans are polysaccharides that exhibit good film forming properties. This work aimed to produce Galactomannan based edible coating for cheddar cheese and its influence on different quality parameters and shelf life of cheese. This work was based on methodology to extract galactomannans from seeds of Cyamopsis tetragonolobus a pod bearing plant of family leguminosae. The galactomannans was extracted by aqueous extraction followed by precipitation with ethanol. Different formulations of edible coatings were used and cheese was evaluated for different shelf-life parameters. It was found that coating with 1.5% galactomannan was effective in giving better attributes to cheddar cheese at 4 C. This work will be of great importance in producing ecologically safe, non-toxic and consumer friendly edible coating. Key words: Edible Coating, Cyamopsis tetragonolobus, Galactomannan, Cheese, Shelf-life INTRODUCTION Use of edible coatings and films is increasing now-a-days because of their certain advantages over synthetic films. They can be helpful to reduce environmental pollution as they degrade easily. Edible films are usually made from substances with film forming properties. These coatings are having ability to improve quality of food and enhance shelf life of edibles to

which they are applied (Mc Hugh and Senesi, 2000).They exhibit better barrier properties to moisture, oxygen and solute movement (Kester and Fennema, 1986; Bourtoom, 2008) and also function as carrier of food additives e.g. anti-microbial, anti-browning and antioxidant agents, flavors, colorants and spices (Pranoto et al., 2005).Plasticizer such as glycerol are added to increase film flexibility and decrease barrier properties (Casariego et al., 2008). Guar beans scientifically known as Cyamopsis tetragonolobus belongs to family leguminosae and sub-family papilonaceae yield guar gum a galactomannan of commercial importance. Galactomannans are heterogeneous polysaccharides, composed of straight back bone of -mannose backbone in (1-4) linkage and a side chain of -galactose in (1-6) linkage ( Srivastava and Kapoor, 2005).Galactomannan forms highly viscous solution at very low concentration ( Sittikijyothin et al., 2005)..There are superabundance uses of guar gum in food and non-food industry because of its spacious functional properties. Guar gum finds its application as a food additive in various food products and employed mainly as emulsifying agent, thickener, stabilizer and gelling agent in food industry (Flammang et al., 2006). Cheese is the generic term used for group of dairy products derived by fermentation process (Fox and McSweeney, 2004).Cheddar cheese is classified as hard ripened cheese and is popular worldwide due to its distinct flavor, aroma and taste .Cheddar cheese is produced by acidification of milk followed by coagulation by rennet (Bank, 2002). It is having a buttery but firm body with close texture and a clean nutty flavor (Varnam and Sutherland, 2001).Ripening of cheddar cheese is a quite perplex chemical and biochemical process that involves degradation of curd constituents by enzymes (Choisy et al., 2000).

Effect of galactomannan based edible coating on shelf life of ricotta cheese was evaluated by Martins & Co. workers (2010).Edible coating made up of galactomannan from Gledtisia triacnthos with nisin incorporated was helpful in retarding the growth of Listeria monocytogenes at 4C ,hence is effective in extending the shelf life. Three different treatments were tested for cheese samples: Samples without coating, Samples with coating without nisin and samples with coating with 50 IU g-1 of nisin incorporated. The results of different tests demonstrated that combined action of nisin and novel galactomannan based coating improves the microbial safety and quality attributes of ricotta cheese and extends its shelf life during refrigerated storage (Martins et al., 2010). This study was designed to address following objectives: 1) To develop eco friendly coating material for food preservation. 2) To evaluate the potential of galactomanns from guar beans to extend shelf life of cheddar cheese. 3) To evaluate the changes in different quality attributes of cheese under comparative storage temperatures. MATERIALS AND METHODS Guar seeds purchased locally Ethanol absolute (Merck) Fresh milk (It will be purchased fresh from milk man) Starter Culture of Lactobacillus acidophilus (From microbiology Lab. Food technology department, UAAR)

Glycerol 87% Corn Oil Extraction of Guar Gum Galactomannan from Guar beans Polysaccharide (Galactomannan) was extracted from guar bean seeds using ethanol and distilled water. Seeds were removed from pods, cleaned and weighed. After weighing seeds were placed in blender to cause their mechanical breakage. Endosperm would be manually separated from germ and the hull. Then endosperm of seeds was in 99.8 percent pure ethanol in 1:3 (Endosperm: Ethanol) at 70C for fifteen minutes to inactivate enzymes and low molecular weight substances. Ethanol was decanted and endosperm was suspended in distilled water in 1:5 for 24 hrs approximately. Water as added to suspension in 1:10 and liquefied using blender for 5minutes (Cunha et al., 2009; Dakia et al., 2008). Polysaccharide Purification The endosperm mixture was filtered and then centrifugation was carried out at 3800 rpm for 20 min at 20C. Supernatant was collected and galactomannan was precipitated by adding ethanol to supernatant (1:2). Ethanol was decanted and galactomannan precipitate was collected and stored at 4 C (Cunha et al., 2009; Dakia et al., 2008). Cheddar Cheese Preparation Milk was pasteurized at a temperature of 70 C for 15-20 sec. Milk was cooled and stored for analysis. Then standardized milk containing about 3.3 % fat was warmed to 30 C in cheese vat. 2 percent starter culture will be added to warm milk to bring pH of milk to 6.2. After about half an hour, when pH of milk reached 6.2, rennet was added at 1ml per 4 lit of milk. After 40-50

min when curd was formed, it was cut into cubes of size 1.25 cm. Curd cubes and whey was heated to 38-40C with continuous stirring. After removing whey curd blocks were gently heated at 30C for 2 hrs for cheddaring process to take place. Salt was added at 2-2.5 percent. Salted curds were placed in mould and pressed manually. Cheese was packaged in polyethylene bags. Cheese was stored at 4 C temperature until treatments were applied (-Connor, 1993). Preparation of Coating Forming Solution Two sets of solutions to form films were prepared by dissolving 1.5 percent w/v and 0.5 percent galactomannan in distilled water at 200 rpm for 24 hrs at room temperature with the help of magnetic stirrer. Corn oil was added at a concentration of 0.5 percent w/v under agitation of 20min at 60 C. At a concentration 2.0 percent w/v glycerol was added. Coating Solution Application Cheddar cheese prepared was sliced into small pieces. The film forming Solution was applied to cheese slices by gently dipping. Samples were allowed to rest for some time to drip off residual solution. Then cheese was placed at 4C for 4hrs until coating was dry. Physico-chemical analysis pH was measured using electrode method by dipping electrode in cheese slurry. Weight loss percentage was calculated by measuring initial and final weights of cheese samples. Moisture content of cheese samples was determined by drying samples in hot air oven at 105C until constant weight of dried cheese was acquired (Martins et a l., 2010). Kjeldhal method was used to determine protein content (Effat et al., 2012). Total solid content of cheese was determined by drying sample to a constant weight in an oven at 105 C for 4-5hrs. (Method

33.2.44; 990.20, AOAC 2006). Gerber method was used to determine fat content of cheddar cheese samples. Organoleptic Evaluation Cheese samples were evaluated for different sensory attributes like color, taste, flavor and aroma by a panel of judges (Effat et al., 2012) Microbiological Analyses Total viable count of cheese samples was done (Effat et al., 2012) Statiscal Analyses Data was analyzed using Microsoft windows excel 2007 and Analysis of variances (ANOVA) was applied using statistical software Statistix 8.1.

RESULTS AND DISCUSSION

PHYSICO-CHEMICAL ANALYSIS OF CHEESE pH A significant decrease in pH values for cheese samples was observed at both the temperatures. Maximum value for pH was 5.12 0.02 shown by samples at 0 day at T0 (Control).The minimum value for samples was 4.96 0.08 exhibited by samples at 21 day of storage having treatment T1 (1.5%).At room temperature pH changed from 5.12 0.02 at 0 day to 4.98 0.05at 21 days of storage for Control for all the samples. Decrease in pH was due to increase in acidity. Increase in acidity was due to more lactic acid production by lactic acid bacteria. Weight Loss A significant weight loss was observed for all the cheese samples. At a temperature of 4 C the highest weight loss value was 22.23 0.02 percent for T0 during 21 days of storage. Lowest weight loss value was 20.16 0.02 percent for T1 and for T2 weight loss value was 21.70 0.26 percent. For samples at 25 C maximum weight loss value was 23.49 0.03

percent for T0. For T1 and T2 weight loss values were 22.10 0.00 percent and 23.16 0.01 percent respectively during 21 days of storage. Moisture Content A significant decrease in moisture content was observed for cheese samples. For samples at 4 C maximum moisture content value was 47.39 1.01 percent for T1at 21 day.T0 exhibited the lowest moisture content value of 45.92 0.34 percent during 21 days of storage. Similarly, a decrease in moisture content was observed for samples stored at room temperature. In case of

uncoated cheese moisture content decrease from 49.56 1.58 percent at 0 of storage to 45.48 0.47 percent at the end of 21 day. At 0day maximum moisture content value was 49.98 1.25 for samples with T1 stored at room temperature. Protein Content An increase in protein content of cheese samples was there. In cheese samples with T1 maximum protein value of 19.13 0.00 percent was exhibited at 21 day of storage. The lowest value was 18.98 0.02 percent possessed by cheese samples without coating T0 at 4 C. At

room temperature maximum value for protein content was 19.10 0.01 percent and minimum value was18.92 0.00 percent exhibited by T1 and T0 respectively. Total Solids Total soluble solids are directly related to moisture content of the cheese. As moisture content decreases total soluble solids increase. Maximum value for total solids was shown by samples without coating T0. Maximum values for total solids were 54.05 0.39 percent and 54.51 0.47 percent for T0 samples at 4 C and room temperature respectively. For samples with T1 and T2 total solids content varied from 49.95 1.16 percent and 49.96 1.25 percent at 0day of storage to 52.60 1.01 percent and 53.44 0.21 percent at 21 day of storage at 4 C. Ash Content Ash content values of cheese samples varied from 2.33 0.00 percent at 0day to 2.46 0.01 percent.

Fat Content An increase in fat content was observed during storage period. Maximum fat content value was 22.10 0.17 percent for T0 at 21 day of storage. At 0 day lowest fat content value was 21.40 0.17 percent at 4 C. At room temperature fat content increased from 21.40 0.17 percent at 0day to 22.20 0.00 percent during 21 days of storage. Organoleptic Evaluation Organoleptic evaluation of cheese samples was done by a panel of seven people. Microbiological Analysis Total viable count values at 4 C were 1.68 0.11 log CFU/g value for T0 during 21 days of storage. This is the highest value for samples stored at 4 C. For T1 and T2 log CFU/g values were 1.66 0.11 and 1.67 0.11 respectively. At room temperature log CFU/g values during 21 days of storage were 1.71 0.11, 1.68 0.11 and 1.70 0.11 for T0, T1 and T2 respectively. Conclusion: It was found that edible coating with 1.5 percent galactomannan exhibited better effect on shelflife parameter of cheddar cheese as compared to samples without coating or those with 0.5 percent galactomannan coating. Galactomannan from guar beans exhibit better coating properties so it can be used as a raw material for galactomannan based edible coatings.

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