Biotechnological Communication

Biosci. Biotech. Res. Comm. 8(2): 138144 (2015)

Comparative biochemical analysis of certain indigenous rice beverages of tribes of with some foreign liquor

J. Arjun* Department of Zoology Lumding College, Lumding Assam 782447

ABSTRACT In north east various types of beverages are prepared by tribal people Viz Poro, Horo Judima and jumai by Miss ing, Karbi, Dimasa and respectively. Carbohydrate, protein and alchohol percentages were estimated both in indigenous beverages and foreign liquor. The local tribal wine show more, protein, carbohydrate than foreign one. The present investigation was undertaken to evaluate the biochemical and nutritional status of Judima, Jumai, Horo and Poro tribal preparations after successive fermentationed of alcoholic beverages. At the same time photochemical analysis of plants used in judima, jumai, horo and poro on PBMC cells were also evaluated to determine their toxic effect on the host if any, consideering ethanol (50% and 100%) as standard. The present investigation clearly showed that indigenous rice beverages contains more nutrient value that of foreign liquor. Thus it can be predicted that care ful identi cation and selection of yeast and bacteria and useful fungi from starter culture employed in traditional brewing in N.E India yield industrially important and bene cial microbes for the bene t of mankind and at the same time industrialization of tribal wine massy be very useful from the physiological point of view.

KEY WORDS: INDIGENOUS, BEVERAGES, FOREIGN LIQUOR, BIOCHEMICAL PROPERTY

INTODUCTION and owers is a common practice among many tribal In north east India, The preparation and consumption communities. Each tribe prepare their own unique cul of rice beers such as poro, horo, jumai and Judima from tures to carry out fermentation and each type is a mix different varieties of rice ingredients with plants, fruits, ture of different parts of various plants (Das et al 2012).

ARTICLE INFORMATION: *Corresponding Author: Received 20th August, 2015 Accepted after revision 16 th October, 2015 BBRC Print ISSN: 0974-6455 Online ISSN: 2321-4007 NAAS Journal Score : 3.48 © A Society of Science and Nature Publication, 2015. All rights reserved. Online Contents Available at: http//www.bbrc.in/ 138 J. Arjun

The household liquor is associated with regions rich photochemical analysis of plants used in judima, jumai, indigenous knowledge system (Achaya 1991). horo and poro on PBMC cells were also evaluated to The traditional knowledge of beverages making prac determine its toxic effect on the host if any, consideering tice in NE region has been transformed from one gen ethanol (50% and 100%) as standard. eration to another through ages, (Tanti et al 2010, Das et al 2012). Fermentation is a natural process in which carbohy MATERIALS AND METHODS drates are oxidized to alcohol and other compounds by anaerobic microbes. It is a biological oxidation proc ess employed by certain microorganism for their energy FIELD SURVEY: requirements (Kuwaki et al 2012). Microorganisms are the chief sources in connecting a particular food sub The eld survey was performed using Sami structure stance in fermented forms. Alcoholic beverage were questioneriess personal visits were made to homes, produced individually in many countries even in 19 th working place and wine selling markets. A face to face century, (Achaya 1991). interview was made in local languages about the method Poro, Horo, Judima and Jumai are the most popular of preparation of judima, jumai, horo and poro in Karbi, homemade alcoholic rice beverages of the tribal com Missing and Dimasa people of Karbi anglong and N.C munities of Assam such as Misshing, Karbi and Dimasa, hills (Dimahasas) district. Boro people.The present study was carried out in North cachar hills(Dima Hasao) and Karbianglong district of Assam. All of the beverages are prepared by fermenta PREPARATION OF JUDIMA, HORO, PORO AND tion of worked rice mixed with plant leaves and barks JUMAI containing different phytochamicals broadly described as phytoestrogens, terpernoids, carotenoids, limnoids, The whole process for preparation of judima, jumai, phytosterols, glucosinolates, polyphenols, avonoides, horo & poro are more or less same. The process involves iso avinoids and anthocyanidins. They have tremen two stages. dous impact on the healthcare system and may provide • Making of rice cakes medical health bene ts including the prevention and or • Preparation of rice beer treatment of diseases and physiological diseases. These photochemical either alone and or in combination have tremendous therapeutic potential curing various ail ments (Prakash et al 2012). PREPARATION OF RICE CAKES For the fermentation of all types of above mentioned The starter cake of judima, jumai, horo and poro is beverages the starter material is homemade starch. Yeast made by the mixture of rice and bark of plants e.g ( culture maintained semi sterilized medium made of Acacia pennata ). For variation in tastes they use mix ground rice (moist gluten) mixed with a number of plant leaves powder of piper betel (leaves), Buddeja asiatica parts are repotted to carry medicinal properties (Das (leave things) and Hedystis scandass ( leaves things). The et al 2012). The fermentation temperature during indig barks of Acacia Pennata are cut in to small pieces and enous process of wine making is generally maintained at dried in the sun. Glutinous rice (Oryza glutinous)f lour is around 30°c. Since the entire practice of beverage prepa soaked in water until it is softened. It is then grinded in a ration has been traditional and consumption of fermen wooden or metallic mortar and pestle called remin along tation forth is a crude product, it was therefore felt that the bark of Acacia pennata plans. A little water in added the evaluation of biochemical status of the consumed in order to make a paste it is then made in to cakes of product is de nitely inevitable. appropriate sizes (radium 57 cm, weight 80100 gr). The The present investigation was undertaken in an old rice cake (1520gr) was then powdered and sprinkled attempt to evaluate the biochemical and nutritional sta over the new rice cake. New rice cake was then it kept tus of Judima, Jumai, Horo and Poro after successive for at list 24 hours an then it is sun dried till it becomes fermentation of alcoholic beverages. At the same time hard and it can be stored for many months.

BIOSCIENCE B IOTECHNOLOGY R ESEARCH C OMMUNICATIONS COMPARATIVE BIOCHEMICAL ANALYSIS OF CERTAIN INDIGENOUS RICE BEVERAGES 139 J. Arjun

PREPARATION OF RICE CAKE

PREPARATION OF RICE BEER For preparing judima ,jumai, horo and poro rice is boiled and allowed it cool. It is mixed with powdered mixture of rice and bark of plant (Acacia pennata ) and kept in a large container which is covered with jute gunny bags. After 57 days slightly yellowish, creamy white, white juices come out from the fermented mass which indi cates the completion of fermentation process. This is further diluted with water and ltered for consumption.

FLOW CHART FOR THE PREPARATION OF RICE BEER

ESTIMATION OF ALCOHOL In brief, 1.0 ml properly distilled sample and 4.0 ml The decanted products of judima,jumai, horo and poro chromic acid was incubated at boiling water for 10 min were centrifuged at 5000 rpm for minutes at 4° c and utes cooled at room temperature and optical density supernatant of which was used for estimation of alcohol. (0.D) was read at 600 km (U –V visible spectrophotom The ethanol content of judima,jumai, horo, poro, beer, eter Beckman, Model DU 640) using chromic acid with gin, vodka, rum and whisky was estimated by chromic distilled water as blank. The alcoholic % of foreign liq acid method (Willium and Darwin1950). uor were also nd out by the same method.

140 COMPARATIVE BIOCHEMICAL ANALYSIS OF CERTAIN INDIGENOUS RICE BEVERAGES BIOSCIENCE B IOTECHNOLOGY R ESEARCH C OMMUNICATIONS J. Arjun

ESTIMATION OF TURBIDITY CARBOHYDRATE (3840°c) triturated and subjected to the maceration AND PROTEIN with 70% methanol (2lt) for seven days with daily agita tion. The extracts were ltered and concentrated under The turbidity of the decanted product was determined reduced pressure in order to obtain methanol extract . against distilled water at 640 by UV visible spectro The methanol extracts of all plants were transferred into photometer (Beckman would DU 140. The total car airtight culture vial with proper labeling for chromato bohydrate of the decanted product was estimated by graphic and phytochemical analysis. Anthrone reagent method by Morris 1940 using 1.0 mg/ ml glucose solution as standard. The O.D reaction mix THINLAYER CHROMATOGRAPHY AND ture was taken at 550 nm against reaction mixture as blank in which glucose solution was replaced by dis PHYTOCHEMICAL ANALYSIS tilled water. The total protein content was determined by The phyto chemical analysis namely phenols, tannins, Folin – phenol method (Lowry etal (2003) The estimation avonoids, saponins, glycosides stepids terpenoids and of all beverages of both indigenous and foreign alkaloids of selected plants used as common growth sup Liquor have been nd out by two above maintained plements during the preparation of fermentation starter process. culture was carried out according to the methods men tioned by Yadav and Agarwale (2011)

COLLECTION OF PLANTS USED FOR THE STATISTICAL ANALYSIS PREPARATION OF JUDIMA, JUMAI, HORO AND PORO All values in the plant study indicates mean and stand ard deviation (S.D) and all determinations were car Fresh parts of form plants piper bettle (Leaves), Buddeja ried out repeated three times. The one way analysis of asiatica (leaves), Hadyotis scandens (leaves) and Acacia variance (ANOVA) was used to evaluate the difference pennata (leaves) are used for the preparation of fermen among multiple groups followed by a post hoc test (Tur tation starter culture from different location of N.C hills key) statistical signi cance was taken a 95% con dence district of Assam. Plant leaves (50g) was dried on oven limit.

Phytochemical constituent of four plants used as common growth supplement during the preparation of fermentation starter culture contrary Brewer’s Yeast.

Plants Proteins Carbohydrate Phenols Flavonoids Saponins Glycosides Steroids Terpinoids Alkaloids Tannins

Acacia ++ ++ ++++ ++ + ++ + + + pennata (0.33) (0.44) (0.32) (0.24) (0.21) (0.45) (0.14) (0.025) (0.19)

Buddleja + +++ +++ +++ + + + + ++ Asiatica (0.21) (0.48) (0.27) (0.29) (0.024) (0.32) (0.11) (0.02) (0.28)

Hedyotis + ++ +++ ++ + + + ++ + scandens (0.23) (0.43) (0.24) (0.22) (0.022) (0.30) (0.14) (0.120) (0.17)

Piper ++ ++ +++ ++++ + ++ ++ + +++ Betle (0.30) (0.42) (0.24) (0.31) (0.27) (0.42) (0.21) (0.27) (0.838)

Judima o.28 0.18 0.23 0.16 _____ 0.20 ______0.11

Horo 0.27 0.28 0.22 0.18 ______0.18 ______0.12

Poro 0.26 0.26 0.25 0.17 ______0.19 ______0.14

Jumai 0.28 0.23 0.24 0.18 ______0.20 ______0.12

BIOSCIENCE B IOTECHNOLOGY R ESEARCH C OMMUNICATIONS COMPARATIVE BIOCHEMICAL ANALYSIS OF CERTAIN INDIGENOUS RICE BEVERAGES 141 J. Arjun

Comparative biochemical analysis of indigenous rice beverages and foreign liquor

Indigenous Wine Foreign Wine

Characteristics Horo Poro Judima Jumai Beer Gin Blue VodkaWhite Ram Whisky King sher Ribbon Mischief McDowell Mc Dowell

Colour White Red Yellesh Creamy Light Transparent Transparent Deep Yellow White Yellow Yellow

P? 5 5.6 4.2 4.4 6.2 6.2 6.3 6.5 6.6

Alcohol% 36% 22.88% 39% 40.1% 9% 42.8% 42.8% 42.8% 42%

Protein 3.2 5.7 4.2 6.2 0.11 0.13 0.17 0.12 0.15 content mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml

Carbohydrate 12.3 14 12 13.2 1.22 1.45 1.96 1.49 1.42 Content mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml

Free amino 0.21 0.22 0.24 0.20 0.09 0.08 0.09 0.08 0.08 acid mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml mg/ml

RESULTS AND DISCUSSION alcohol during domestic alcohol fermentation occurs in two phases viz sacchari cation of rice to simple mono The color of various indigenous beverages one white and disaccharides through hydrolysis of starch and con (Horo), red (Poro), judima (Yellowsh), jumai (creamy version of monosaccharides to alcohol. The sacchari ca white). The color of the foreign liquor such as Beer (light tion process is accomplished by extracellular enzymes yellow), Gin (transparent), Vodka(transparent), Rum ? amylase and Eglucosidase which has been observed (deep yellow), whisky(yellow). The p ? of the indigenous to be expressed in many yeast strains. The quality of wine such as Horo, Poro, Judima and Jumai are 5, 5.6, amylase (E.C.3.2.1.1) which hydrogen D14 glycoside 4.2, 4.4 and that of foreign liquor such as Beer, Gin, linkages of starch to produce maltose and other disac Vodka, Rum and Whisky are 6.2, 6.2, 6.3, 6.5 and 6.6. charides in the decanted product is found to be more The alcoholic % of indigenous wine showing lesser prominent (4.76 u/ml/min). Glucoamylase activity (E.C than that of foreign liquor i.e alcoholic % of Horo, Poro, .3.2.1.1.)Which hydrolyses D 14 and D 16 glycosidic Judima and Jumai are 30%, 22.88%, 30% & 40% but linkages to form glucose. Alcohol fermentation succes alcoholic % of foreign wine are Beer(9%), Gin (42.8%), sively involves different microorganism although yeasts Vodka (42.8%), Rum (42.8%), Whiskey (42%). The pro are the most prominent species observed being the the tein content of indigenous beverages Horo,(3.2 mg/ml), reason for such different activity of D amylase and glu Poro (5.7 mg/ml), Judima(4.2 mg/ml), Jumai(6.2 mg/ml) coamylase enzyme,(Bnyzak 2003). where as protein content of foreign liquor Beer (0.11 mg/ On the other hand, the use of plants, materials and ml), Gin(0.13 mg/ml), Vodka (0.17 mg/ml), Rum (0.12 plant materials and plant root extracts such as Old- mg/ml), whisky (0.15 mg/ml). dendia, corymbosa, Cyclons sp, Adhatoda vasica, Zan- The carbohydrate content of indigenous beverages thoxylum hemitinianum, Dracere angustifolla, Naravelia Horo(12.3 mg/ml), Poro (14 mg/ml), Judima (12 mg/ feliavica, Melothrea Sp as essential ingredients during ml), Jumai (13.2 mg/ml) when a carbohydrate content preparation of starter Culture for the fermentation of foreign beverages are Beer (1.22 mg/ml), Gin (1.45 mg/ alcoholic beverages (Tamang et al , 1996) may contribute ml), Vodka (1.96 mg/ml), Rum (1.49 mg/ml) and Whisky to addition to phenolics, which in turn might have con (1.42 mg/ml) tributed to changes in microbial population might have The presence of carbohydrate in decanted product added to Starter cultures along with plant ingredients. can be added for the energy metabolism in the cells. Such results obtained in the present ndings suggest The presence of protein will act as metabolisable and that the microbial consortia involved in the fermenta energy providing substance in the decanted product, tion of the said alcoholic beverage indeed is the re ec which would certainly enhance the nutritional status of tion of their microbio logical and biological character the beverages. The biochemical conversion of starch to istics (Black 1999 & Sharma 2002). Although isolation

142 COMPARATIVE BIOCHEMICAL ANALYSIS OF CERTAIN INDIGENOUS RICE BEVERAGES BIOSCIENCE B IOTECHNOLOGY R ESEARCH C OMMUNICATIONS J. Arjun

of various amylolytie yeast from starter cultures has preparation of “Tapeketan”. The growth of various yeast been reported earlier. Yet it was found that all of these has been shown to be dependent on amylolytic capacity were not responsible for predominance during fermenta of the mould given rice to typical avor of ‘Tapeketun’. tion (Tsuoshi et al 2004). It has been reported that two In recent yeast people have taken interest in traditional types of yeast are involved in ‘Jaan’ fermentation in Sik medicines derived from plants. The people of kim white amylolytic yeasts from degrade starch & pro use their traditional liquor (yu) for medicine, relaxant, duce glucose (Tsuyoshi et al 2004) the alcohol producing and of offerings such which is a dilled product of fer yeast from then groes rapidly on the resultant glucose to mented local rice. It is a strong solvent for many impor ethanol (Fleet et al 1993). tant active constituent of medicinal plants whose action The biochemical parameters of carbohydrate protein plays a vital role in traditional medicine (Singh & Singh, and alcohol % P ? color etc content of foreign liquor 2006) similar observation also been reported regarding wine estimated the protein carbohydrate content of for the local liquor called ‘Rokshi’ of (Singh et al eign shoes less than tribal wine. The total carbohydrates 1999, Singh & Singh 2006). protein and free amino acid content in decanted product The prevalence of traditional food in the state in of judima, horo, poro and jumai was estimated the pres largely linked to availability of raw materials, geograph ence adds to the energy metabolism in the cell. The pres ical barris environmental conditions and different ethnic sure of these metabolism and energy processing sub tribal groups (Savitri et al ). In Manipur traditional food stance in the decanted product would certainly changes & beverages are consumed as regular foods (Jairam etal the nutritional status of the beverage which is consumed 2009). Mutherian tribal community of Iduki district of as refreshing drink. Kerala palm wine and consumed directly without fer The biochemical conversion of starch to alcohol mentation (Monithorum et al 2007). Kiad a popular local during domestic alcohol fermentation occur in two liquor of pnar tribe of Jaintia hills in consid phases viz sacchari cation of rice to simple mono and ered to be good for health and acts as remedy for various disacchari cation thought hydrolisic of starch and ailments such as Urinary bladder, dysentery (Sowati et conversion of monosaceharides to alcohol. The saccha al 2007) ri cation process is accomplished by the extracullu Apart from experiments performed and result lar enzymes D amylase & E glucosidase (Bryzak 2003), obtained, it can be predicted that careful identi cation which has been observed to be in many yeast steains & selection of yeast and bacteria and useful fungi from and moulds. The amylase in hydrolysis of 14 glycoo starter of cultures employed in traditional brewing in side linkage of starch to produce to maltose and other N.E India can yield industrially important and bene disaharides (Bryzak 2003). Alcoholic fermentation suc cially microbes for the bene t of mankind. cessively involves different microorganism although The present investigation clearly showed that indig yeast are the prominent speies. On the other hand the enous rice beverages contains more nutrient value that use of plant materials such as Buddeja asiatica (leaves), of foreign liquor. Thus it can be predicted that careful Hedyotis scandens (Leaves and Twigs) and Acacia pen- identi cation and selection of yeast and bacteria and nata (leaves and barks) as essential ingredients during useful fungi from starter culture employed in traditional preparation of starch for the fermentation of alcoholic brewing in N.E India yield industrially important and beverage may contribute to additional to phenolics bene cial microbes for the bene t of mankind and at which in turn might have contributed changes in the same time industrialization of tribal wine may be microbial population during preparation. Recent report very useful for the physiological point of view. for alcoholic fermentation of Manipur shows Hamic as starter, traditional fermented foods of Naga tribes (Mao and Odoiso 2007) and Sajan, a slocal rice beer of Deori REFERENCES tribe (Deori et al 2007) suggest the use of starch in the Achaya, K.T. 1991. 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BIOSCIENCE B IOTECHNOLOGY R ESEARCH C OMMUNICATIONS COMPARATIVE BIOCHEMICAL ANALYSIS OF CERTAIN INDIGENOUS RICE BEVERAGES 143 J. Arjun

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144 COMPARATIVE BIOCHEMICAL ANALYSIS OF CERTAIN INDIGENOUS RICE BEVERAGES BIOSCIENCE B IOTECHNOLOGY R ESEARCH C OMMUNICATIONS