Mango Wine: Standardisation and Screening of Cultivars Using Different Strains of Saccharomyces Cerevisiae

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1 Asian Journal of Multidisciplinary Studies Volume1, Issue 4, November 2013 ISSN: Available online at Mango Wine: Standardisation and Screening of Cultivars Using Different Strains of Saccharomyces Cerevisiae Vimala Beera 1, Ravi Babu Mangam 2, and Kiranmayi Enthoti 1 Research Scholar, IARI, New Delhi 2 Assistant professor, ANGRAU, Rajahmundry Abstract : Preliminary screening of six mango varieties viz., Baneshan, Totapuri, Cherukurasam, Mallika, Manjeera, Suvarnarekha with two yeast strains of Saccharomyces cerevisiae (MTCC 172 and AM 113) for wine making was undertaken. The wine produced from these cultivars was analysed for various bio chemical properties at 45 and 90 days after storage. Wines produced from cv. Baneshan with 172 were better when compared with Saccharomyces cerevisiae AM 113 among all qualitative aspects. The quality of wines from all the cultivars at 90 days of storage was found to be high irrespective of the yeast strain. Based on the alcohol content, total carotenoids and tannin content, the wines produced from cv. Baneshan with Saccharomyces cerevisiae MTCC 172 is recommended for wine making from mango. Keywords: saccharomyces cerevisiae, amelioration, mango cultivars, value added product, wine INTRODUCTION India is the second largest producer of fruits in the world after China with a production of million tonnes. Mango (Mangifera indica L.) ranks first in area and production covering an area of 2309,000 hactares with a total production of 12,750,000 tonnes per year (NHB, 2009). Fruits are perishable and the overriped fruits cannot be transported to long distances and when there is glut in the market, the prices fall down and the fruits will be available at cheaper rate. Further, the undersized and irregularly shaped fruits that are available at lower price in the market can be converted into various value added products including wine. Wine is a fermented beverage from fresh fruits composed of water, alcohol, pigments, esters, vitamins, carbohydrates, minerals, acids, tannins with medicinal and therapeutic value (Patil et al,2005).wines have always been considered as safe and healthy drinks, besides an important adjunct to the diet. In wines, ethanol is a macro nutrient energy source, capable of providing calories for all essential biological activities of the human cells, energy for physical work and thermogenesis (Bisson, 1995). Grape has been the principal fruit employed in the preparation of a variety of wines (Vyas and Chakravorthy, 1971). Though several fruits are used for wine preparation, they were usually found inferior to the grape wine. Mango is decidedly the choicest of all fruits because it possesses attractive colour, highly acceptable taste and excellent flavour (Srivastava, 1998). Mango is processed into various products such as slices, pulp, juice, nectar, squash, 35

2 jam, fruit bar, pickle, chutney, vinegar, amchur, wine etc. (Singh and Dhawan, 1983). Mango in general contains high amounts of total soluble solids (TSS), vitamins, minerals thus making it a suitable medium for the growth of wine yeasts. Also it can complement white wines which need to be prepared from white grapes, which in turn are a great worth when processed as raisins rather than wine. Keeping the above facts in view, the present study was undertaken to standardize wine preparation from mango. MATERIALS AND METHODS The materials and the methods followed during the course of experiment are presented here under. 1. Location of the experiment: The experiment was conducted at the Department of Horticulture, Department of Agricultural Microbiology and Bioenergy College of Agriculture and Post Graduate & Research Centre, ANGRAU, Rajendranagar, Hyderabad. 2. Procurement of the materials A. Fruits: Six mango varieties viz., Baneshan, Totapuri, Cherukurasam, Mallika, Manjeera and Suvarnarekha used in the experiment were obtained from Agricultural Research Institute farm, Rajendranagar, Hyderabad. Well matured fruits of uniform size were used in experiment. B. Chemicals: All chemicals used in experiment and analysis were of analytical grade, purchased from standard Indian chemical companies. The dehydrated media and chemicals of HiMedia were used for growing and maintenance of yeast culture. C. Cultures: Two Saccharomyces cerevisiae strains MTCC 172, and AM-113 were obtained from Institute of Microbial Technology, Chandigarh (IMTECH). The cultures were maintained in growth medium No.5 (Agar: 20 g, Dextrose: 20 g, Peptone: 10 g, Yeast extract: 20 g, Distilled water 1L).The medium was maintained at ph The culturing of yeast was done at 30 o C by incubating for 48 h under aerobic conditions. 3. Extraction of pulp & preparation of the must: Five kg each of sound, healthy, well matured and uniformly ripened fruits from six varieties were washed thoroughly, peeled and the pulp was separated from stones using stainless steel knife under hygienic conditions. The pulp obtained from six varieties was homogenized separately in the mixer then added with distilled water at 1:1 ratio and the pulp was ameliorated to 24 o Brix by adding cane sugar as required. To find out Brix values, ERMA hand refractometer was used. Potassium metabisulphite (KMS) was added to the juice at the rate of 200 mg/l for obtaining a final concentration of 100 ppm sulphur dioxide to suppress the activity of various micro-organisms before inoculating yeast strains. The pulp was incubated for 8 12 hrs at room temperature. Asian Journal of Multidisciplinary Studies, 1(4) November,

3 4. Preparation of inoculum: At first, freeze dried cultures were inoculated into the liquid broth separately to facilitate growth. After sufficient growth a loopful was streaked on the agar medium. After verifying the purity of the cultures, they were inoculated into agar slants of the growth medium and incubated at 30 o C for 48 h. Ten ml of sterile liquid medium was transferred to each agar slant culture with a sterilized pipette. Each of the yeast growth was harvested and inoculated into small amount of fresh sterile liquid medium separately which was incubated at 30 o C for 48 hrs to get turbid culture. This turbid culture thus obtained was transferred into large amount of fresh sterile liquid medium separately which was incubated at 30 o C for 48 hrs, to get mother culture. The mother culture was used for inoculation of the must for fermentative production of wine. 5. Inoculation of the must: After holding for 8 12 hours, the must was inoculated with the mother cultures obtained from the strains of Saccharomyces cerevisiae i.e., MTCC 172 and AM 113 in the ratio of 1: 10 under aseptic conditions and left for fermentation at room temperature for 7 days. 6. Clarification and filteration: After completion of fermentation, clear liquid was filtered through a clean muslin cloth and stored in sterile glass bottles at o C for a month until the suspended matter settled leaving clear wine on the top. The wine was further treated with bentonite at the rate of 0.1 percent and left undisturbed for 2 days for clarification of mango wine. After the settlement of yeast cells at the bottom, fermented must supernatant was siphoned out and transferred into fresh sterile bottles. 7. Maturation and storage: The samples were then left for maturation and stored at room temperature for a period of 45 and 90 days. 8. Chemical analysis: The pulp and wines developed were subjected for analysis of various parameters viz; Total soluble solids by using ERMA hand refractometer (Ranganna,1979), Total sugars and Reducing sugars were estimated by Nelson Somogyi method (Ranganna,1979), Titrable acidity (AO AC, 1970), ph by using ph meter, Colour was estimated colorimetrically at 450 nm in terms of total carotenoids, Alcohol content of wine was estimated spectrophotometrically at 600 nm as per the procedure given by Natu et al. (1986). 9. Statistical analysis: The data were subjected to statistical analysis as per the procedure outlined by Panse and Sukhatme (1967). The design adopted was completely randomized design with factorial concept. RESULTS AND DISCUSSION In the present study, the wines thus produced were subjected to analysis for various physico-chemical, microbial and organoleptic parameters at 45 and 90 days after storage. Each of the parameter studied during fermentation and storage are presented here under. Asian Journal of Multidisciplinary Studies, 1(4) November,

4 Asian Journal of Multidisciplinary Studies Volume1, Issue 4, November 2013 ISSN: Available online at Effects on physico-chemical qualities of wine FIG. 1 Effect of yeast strains on quality of stored wines prepared from different mango cultivars (n=3) TSS ( B) TOTAL SUGARS (%) ACIDITY (%) ph ALCOHOL (%) TOTAL CAROTENOIDS (mg/100 ml) 38

5 Table 1: Chemical composition of mango cultivars Varieties Baneshan Totapuri Cherurasam Mallika Manjeera Suvarnarekha Total sugars (%) Reducing sugars (%) ph Acidity (%) Carotenoids(µg/100g) A. TSS: The TSS in the fresh pulp of mango ranged from o B (Table 1). In all the treatments a decreasing trend of TSS was recorded after 45 days of storage which ranged from o B and o B with AM 113 respectively (Fig 1). Thus 70 per cent of TSS was utilized during fermentation. However, 172 was found to be more efficient in utilizing sugars and converting them into alcohol. Storing of wines to 90 days further resulted in decline of TSS ranging from o B and o B with AM 113 respectively. This decrease in TSS might be due to the complete utilization of sugars. And also, with the increase in fermentation time a significant fall in TSS was observed which is maximum in cv. Baneshan (1.00 o B) followed by cv.suvarnarekha (1.30 o B), cv.totapuri, cv. Mallika, cv. Manjeera (1.50 o B). The lowest decrease in TSS was recorded with cv.cherukurasm (2.00 o B). These findings are in accordance with the results of Attri et al. (1994) in sand pear wine. The lower content of TSS in wines indicates the utilization of sugar present in the must during fermentation (Mir et al., 1988). B. Total sugars: The total sugars ranged from (14.8 % %) in fresh pulp of different mango varieties. Upon fermentation into wine there was a sharp decline in total sugars in different varieties which ranged from and per cent with AM 113 respectively for different varieties (Fig.2). This was due to higher rate of fermentation of sugars into alcohol and also greater utilization of sugars by yeast. Further storing of wines resulted a little decrease in total sugars which ranged from ( %) and ( %) with AM 113 respectively. This could be due to the availability of food, favourable acidity and ph and efficiency of the yeast to consume sugars which is maximum in cv.baneshan (1.82%) followed by cv.suvarnarekha (1.96%), cv.totapuri(2.03%), cv. Mallika(2.15%), cv. Manjeera Asian Journal of Multidisciplinary Studies, 1(4) November,

6 (2.14%) and cv.cherukurasm (2.37%). C. Titrable acidity: The titrable acidity of the wines from all varieties increased significantly upon fermentation at both the storage periods(fig.3). This could be due to the production of certain organic acids by yeast. Further storage of wine upto 90 days resulted a marked decrease in the acidity. Potty et al. (1978) also reported increase in acidity of wines produced. The increase in acidity in the wines could be due to production of acids. Beech and Carr (1977) reported that during the fermentation of apple juice yeasts formed organic acids such as malic acid, lactic acid and succinic acids. Othman Abdul Samah et al. (1992) reported that other organic acids such as oxalic, tartaric, lactic, malic, citric and succinic acids were produced during fermentation. D. ph: The ph decreased significantly in wines developed from all varieties of mango upon fermentation at 45 days after storage. The ph in wines of different varieties of mangoes ranged from 2.83 to 3.22 and 2.99 to 3.31 with Saccharomyces cerevisiae MTCC 172 and Saccharomyces cerevisiae AM 113 respectively. Further storage of wines of different mango varieties to 90 days showed a marked decrease in ph which ranged from 2.64 to 2.87 and 2.61 to 2.91 with AM 113 respectively (Fig.4).This is due to further increase in titrable acidity in wines. The decrease in ph in different wines is due to increase in acidity. These results are in conformity with the findings reported by Patil et al. (1995) in Ber wine. E. Alcohol: A significant difference in the alcohol content of the wines produced with the two yeast strains of Saccharomyces cerevisiae was observed with different mango varieties.the alcohol content of the wines produced with different mango varieties with the yeast strains Saccharomyces cerevisiae MTCC 172 and Saccharomyces cerevisiae AM 113 ranged from 9.56 to per cent and 8.90 to per cent respectively at 45 days after storage (Fig.5). The pulp might have contained sufficient nutrients for the yeast to grow and produce alcohol. This variation in the study revealed that the strain 172 was more efficient in the production of alcohol compared with Saccharomyces cerevisiae AM 113.The alcohol content of the wines showed a further appreciable increase upon storage to 90 days, because of the conversion of sugars present in the must by yeast. The alcohol content ranged from 9.96 to per cent and 9.56 to per cent with Saccharomyces cerevisiae MTCC 172 and Saccharomyces cerevisiae AM 113 respectively. Among the different varieties of mango, Totapuri recorded the lowest alcohol content (9.75 %) while Baneshan recorded highest alcohol content (12.38 %). Further the cvs. Mallika and Manjeera (11.8 % and %) recoded higher alcohol than Asian Journal of Multidisciplinary Studies, 1(4) November,

7 Suvarnarekha (11.21 %) and Cherukurasam (10.07 %). Thus, the efficiency of the yeast strains to convert the sugars into alcohol during the fermentation upon storage showed an inclination. These results are in conformity with data reported by Ayoga (1999) in pineapple wine. F. Total carotenoids: The total carotenoids ranged from 2685 to 5071 µg/100 g in the fresh pulp of different mango varieties. Upon fermentation into wine, there was a significant decrease in total carotenoids in different varieties which ranged from 996 to 1872 µg/100 ml and 993 to 1864 µg/100 ml with the two yeast strains of AM 113 respectively at 45 days after storage (Fig.6). The decrease was due to the fact that the yeast might have utilized the carotenoids during fermentation. Further storing of wines of different mango varieties resulted a little decrease in total carotenoids which ranged from 865 to 1867 µg/100 g and 859 to 1861 µg/100 g with AM 113 respectively.this could be due to utilization by the yeast for its survival. G. Tannins: The tannin content of wines produced from different varieties of mango ranged from 0.01 to 0.02 and to with Saccharomyces cerevisiae MTCC 172 and Saccharomyces cerevisiae AM 113 respectively at 45 days of storage.further storing of wines resulted a marked increase in tannin content. The increase was maximum in cv. Cherukurasam (0.027) and minimum in cv. Baneshan (0.013). The increase in tannins content upon storage was due to complexing of tannins with proteins (or) polymerization and subsequent precipitation. Also the decrease was due to utilization of more amounts of tannins by the yeast cells. The results obtained are in conformity with that obtained in mango wines by Kulkarni et al. (1980). CONCLUSION From the present experiment, it could be concluded that between the two strains, 172 was found to be more efficient for the production of wine with higher alcohol content for different mango varieties. And cv. Baneshan is best best suited for wine production from mango with 172. REFERENCES Ali Simon, R. and Badrie, N. (2002). Utilisation of peel in plantain wine production. Journal of Food Science and Technology Pp Amerine, M.A., Berg, H., Kunkee, R., Ough, C., Singleton, V. and Webb, A. (1980). Technology of wine making. AVI publishing Co, Westpost, Connecticut, USA. AOAC. (1970) Official methods of analysis of the AOAC Washington D C 11 th Edition Pp Asian Journal of Multidisciplinary Studies, 1(4) November,

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