African Journal of Food Science Vol 5 (16), pp. 851-860, 23 December, 2011 Available online at http://www.academicjournals.org/AJFS DOI: 10.5897/AJFSX11.008 ISSN 1996-0794 2011 Academic Journals Review Status of wine production from guava (Psidium guajava L.): A traditional fruit of India Gurvinder Singh Kocher* and Pooja Department of Microbiology, Punjab Agricultural University, Ludhiana- 141004, India. Accepted 29 November, 2011 Thepotentialofwineproductionfromguavaispresentedinthisreview.Guavaiseasytoculture, posseseshighnutritivevalueanditsproductslikejuices,beverages,nectarsetc.arelargely appreciated by the consumers. Guava juice requires chaptalization so as to adjust its Brix and prepare aperfectwineoutofit.Thechaptalizedjuice(must)istreatedwithpectinaseoracombinationof enzymes and fermented with traditional yeasts at a temperature range of 22 to 30C and inoculum size of 6 to 11% (v/v). The addition of N and P improves ethanol production and quality parameters of guava wine. Racking and ageing of guava wine also improves the sensory and organoleptic characteristics of guava wine. Key words: Guava, Saccharomyces cerevisiae, Brix, inoculum size, temperature, DAHP supplementation. INTRODUCTION The nutritional roleof wine is important since its average contribution to total energy intake is estimated to be 10 to 20% among adult males (Macrae et al., 1993). During the pastfewdecades,grapeshavebeenthemainfruitfor wineproduction.Despitethat,severalstudieshave investigated the suitability of other fruits as substrates for thepurposeofwineproduction(Okunowoetal.,2005; Joshi and Attri, 2005). Moreover, the seasonal availability andhighcostofgrapesinthetropicalregionshasalso necessitatedthesearchforalternativefruitsourcesin tropical countries (Alobo and Offonry, 2009).ThereisabundanceoftropicalfruitsinIndiawhich includesguava,watermelon,pineapple,plum,orange etc.Thesefruitsarehighlyperishable,beingsusceptible tobacterialandfungalcontamination,thusleadingto theirspoilage,mechanicaldamageandoverripeness (IhekoroyeandNgoddy,1985).Hence,thesefruitsare difficult to keep for long and are utilized either as fresh or processed into juice and speciality products (Oyeleke and Olaniyan, 2007).Highratewastageofthesefruitsespeciallyattheir peakofproductionseasonnecessitatestheneed for *Correspondingauthor.E-mail:g_kochar@yahoo.comor gskocher@pau.edu.Tel:0091(161)2401960-79ext.330.Fax: 0091(161)2400945. alternativepreservationandpostharvesttechnologies towardstheirvalueadditionthatcanreducethelevelof postharvestlossesbesidesincreasingdiversityofwines (Okoro, 2007; Alobo and Offonry, 2009). GUAVA AS A WINE SUBSTRATE Guava (Psidium guajava L.) (Family Myrtaceae) is one of themostimportantfruitsinIndia.Itisoneoftheexotic fruits prized for its very pleasant, sub acidic and aromatic pulp.Guava,knownasthepoorman'sappleofthe tropics,hasalonghistoryoftraditionaluse,muchof which is being validated by scientific research. It is fourth mostimportantfruitinIndiaaftercitrus,mangoand bananaandsecondinthestateofPunjabafterkinnow withanannualproductionof1.5lakhMT(Stat.abs.Pb, 2009).Itrankssixthintermsofacerageunderdifferent fruitsinthiscountryandisgrowninthestatesofUttar Pradesh,Bihar,MadhyaPradeshandMaharashtraat large scale. Guavahasgreatpotentialforextensivecommercial usebecauseofitseaseofculture,highnutritivevalue andpopularityofprocessedGuavaproducts.Whileripe fruitisusuallyeatenasadessert,processedproducts likejuices,nectar,jam,jellies,babyfoods,puree, beveragebase,syrupandwinearealsopreparedfrom guava (Shankar et al., 2006).852Afr. J. Food Sci. Somepartsofguavatreehavemedicinalusesasits leaves are used for curing diarrhea. Leaves can be made intoteaandthisastringentdecocationcancure stomacheandactasavermifuge(Praneeetal.,1999). Fermentedguavafruitisalsohelpfulincuringdiabetes (Kavimani et al., 1997). Besides, Guava is a good source ofvitaminC,carbohydrates,proteins,minerals,pectin, calciumandphosphorus(Gargetal.,2007).Thefruit contains high concentration of vitamin A (200to400IU), ascorbicacid(88.2to250.8mg/100g),lycopene(45.3 g/gFW),totalsugars(10to15.3%),reducingsugars (2.05 to 6.08%), acids (10 to 15.3%), pectins (0.62%) and phenols (170 to345 GAE/g FW) (Kaur et al., 2009). CHEMICAL CHARACTERISTICS OF GUAVA Total soluble solids Solublesolidsarethebasicrequirementforthefunction ofguavaoranyotherfruitforwineproduction.Ithas been reported that in white and pink seedling varieties of guava,thechiefsugarcomponentsarefructoseand sucrose(MowlahandItoo,1982).TheTSScontentin differentvarieties(Hafsi,Applecolor,Allahabad Safeda, Sardar, Red fleshed guava, L-49) varies from 8.0to15.0%(Chatterjeeetal.,1992;Singh,1998; Pandey and Singh,1998).Sharma et al. (2010) reported thatthetotalsolublesolidsrangedfrom9.4to13.5Bin 22 genotypes of guava.Individualsugarconcentrationhasbeenfoundto increasegraduallywithfruitgrowthanddevelopment except during the end of the growth period (Rodriguez et al.,1971).Themaximumlevelisfoundtobevaryfrom 5.64 to 7.67, 1.90 to 8.00 and 6.20 to 7.78 mg/ 100 ml of juice for fructose, glucose and sucrose, respectively, in all the cultivars studied by Rashida et al.(1997). In 126 days oldcultivarsofShambati,Pakistani,Shendiand Ganib,fructoserepresented20,38,37and41%, glucoserepresentedaround59,23,25and14%and sucroserepresentedaround21,39,37and45%ofthe totalsugarforthecultivars,respectively(Sharmaetal., 2010).Wefoundreducingsugarscontentof3.40,3.29 and 3.68% in Punjab pink, Arka Amulya and Lucknow-49, respectively(Pooja,2011).TheTSSandfermentable sugarspresentindifferentcultivarsofguavaavailablein literature,thusindicatethatitrequireschaptalization (supplementationofsugars)beforeitisusedforwine production. Titrable acidity Titrable acidityis an asset for the fruits destined for wine production as it provides a conducive environment tothe yeasts. Guava is a sub acidic fruit as its soluble solids are composedmainlyoforganicacidsandsugars.The guava pulp possesses a total acidityof0.3 to 0.8% (w/v) ingeneralwithinterspeciesdifferences.Inguava cultivars(SpearAcid,HisarSafeda,Lucknow-49, Patillo,Punjabpink,ArkaAmulya)acidityhasbeen foundtobeintherangeof0.37to0.96%(Jainand Nema,2007;Sharmaetal.,2010;Pooja,2011).Aulakh (2004) found that the amountof titrable acidityincreases continuously from 30 days after fruit set to 135 days after fruitsetinwinterseason(0.31to0.62%).Similarly, duringrainyseasonitwasfoundtoincreaseupto0.58 from0.28%at110daysafterthefruitset.Theorganic acids(0.3to0.8%)contributingtoacidityinGuavaare mainly citric, malic, glycolic, tartaric and lactic acids (Hui, 2006).Further,aciditywasfoundtobeseason dependentwithAllahabadSafedaandSardarguava cultivarsshowing0.25and0.19%duringrainyseason and 0.31 and 0.33% during winter season crop.The major organic acids in guava pulp have been found tobecitric,malic,glycolic,aceticandlacticacids. ArchanaandSiddiqui(2004)foundthataceticacid rangedfrom55.40to122.13molkg-1.Quantitative determinationsusingsuccinicacidasaninternal standardshowedthatcitricandmalicacidarechief organic acids with lactic acid in smaller quantities among cultivatedguavas.However,inwildguavas,citricacid wasthepredominantacid,withlesseramountsofmalic andlactic acids (Chan etal., 1971). Fifty one acids have beenidentifiedinguava(P.guajavaL.)with(E)-cinnamoicacid(0.4mg/kg)and(Z)-3-hexenoicacid(0.2 mg/kg) as major constituents (Idstein et al., 1985). Ascorbic acid VitaminC(L-ascorbicacid)isthelactone2,3-dienol-L-gluconic acid and it belongs to the water soluble class of vitamins. Humans need a daily intake of 30 to 45 mg per daywhiledeficiencyofthisvitaminleadstoscurvy. Though,orangesarewellknownasfruitsrichinvitamin C but guava is far superior with vitamin C content three to six times higher than that in orange.Also,Whitefleshedguavaisreportedtobeabetter sourceofvitaminC(142.6mg/100g)thanpinkfleshed guavaandisalsorichinotherantioxidantssuchas phenolicsand-carotenes.ThevitaminCcontentvaries widely depending on the cultivar as ascorbic acid content of guava cultivars ranges from 149.0 to 250 mg per 100 g ofpulp(PandeyandSingh,1998;BalandDhaliwal, 2004; Thaipong and Boonprakob, 2005).Thaipong et al. (2005) observed highest vitamin C content in Fan Retief (397 mg/100 g FW). In our studies, ascorbic acid content rangedfrom169.7to229.6mg/100gDWinthree varieties of guava (Pooja, 2011). Pectin Guavacontainshighcontentofpectin(astructural heteropolysaccharidecontainedin the primary cell walls ofterrestrialplants),celluloseandhemicellulose.Pectin contentrangesbetween0.47to100%indifferent varieties(Allahabadsafeda,Banarsisurkh,Lucknow-49,ShambatiandShendi)ofguava(Rodriguezetal., 1971,1997;Pandaetal.,2009).Further,totalpectinfor differentcultivarssignificantlyincreaseswithfruitgrowth anddevelopment.InPakistaniandGanibcultivars,it was found to reach its maximum when the fruits were 106 days old. High content of pectin causes problems in juice extractionwhichresultsinunclarifiedjuiceleadingto hazinessinproducedwine.Hence,toincreasethejuice extractionandforproductionofclearwine,apectinase enzyme pre-treatment is necessary. PREPARATION OF WINE FROM GUAVA Pre-fermentation treatment Commerciallyavailableenzymeshavebeenwidelyused in the oenological industry in wine- producing countries to improveimportantcharacteristicsofwines,suchas aromaandcolour.Theyareusedtoincreasethegrape mustyieldduringpressing,facilitatethesettlingof musts,andimproveclarificationandfiltration.Theuse ofpectolyticenzymeshasbeenshowntobesuitableto improve the extraction of colour in red wines (Revilla and Gonzlez,2003;Bautista-Ortnetal.,2005),aroma compounds(Cabarogluetal.,2003)andsoluble polysaccharides(Docoetal.,2007)fromtheskinsand pulpofthegrapes.Oneofthemoststudiedandwidely used commercial pectinases is polygalacturonase.Commercialpreparationscontainingpectinases, arabinaseandcellulasehavealsobeenemployedfor guavajuiceextraction(Kashyapetal.,2001).Besides, pectinaseshavealsobeenusedtoclearthehazein finished wines and hence clear the wine.Ahmadetal.(2009)reportedthatmaximumyield (85.1%)andclarity(93.5%)wereobtainedusing0.245% pectinase,0.135%cellulaseand0.13%hemicellulaseat 50Cfor9.25hincubation.Asimilareffectwasalso reportedbyDiwanandShukla(2005)inenzymically hydrolyzedGuavapulpunderdifferentsetofconditions. Kauretal.(2009)recommendedoptimizedenzymatic treatmentconditionsasenzymeconcentration0.70 mg/100gGuavapulp,incubationtime(7.27h)and incubation temperature of 43.3C.Pectinaseshavealsobeenshowntobeaffectedby temperature andtime of enzyme treatment as increasing exposure time elevates yield but also causes a reduction inascorbicacidcontentofthejuiceduetoitsoxidation (Imungi et al., 1980). Thus, pectinase has been shown to affecttheoenologicalpropertiesofwines.However,the effectofpectolyticenzymesusingmustfromsun-dried grapesofthePedroXimenezvarietyonoenological parametersbeforeandafterenzymatictreatmentswith pre-fermentative maceration at room temperature forKocher and Pooja853 threehoursrevealedthattheenzymetreatmenthadno effectontotalpolyphenolsandotherchemical characteristics (Espejo and. Armada 2010). In guava, we have seen that a temperature of 45C, for 6 h is sufficient toyielda47%clarityinjuicesofdifferentvarietiesof guava studied (Pooja, 2011). Selection of yeast Yeastasagroupofmicroorganismshasbeen quantitativelyandcommerciallyexploitedasa fermentativespeciestocarryoutalcoholicfermentation andthishasurgedmanyscientiststostudythefactors governingitsgrowth,survivalandbiologicalactivitiesin differentfruitecosystems(HeardandFleet,1985). Yeasts play a prominent role in wine fermentations, which canstronglyaffectthequalityandflavourofthefinal product(QuerolandFleet,2006).Yeastowesits invertingandfermentativepropertytothevarious enzymespresentinitlikesucrase,zymase,maltase, lactase, reductase, carboxylase etc. But yeast of different speciesdonotcontainthesameenzymesandhence differentyeastspeciesbehavedifferentlytowardsthe varioussugars.Amongseveralyeasts,Saccharomyces cerevisiaeandS.bayanusvar.uvarumarethemost importantspeciespresentduringthefermentation process (Pretorius, 2000;Querol and Fleet,2006). It has beenestablishedthatthegrowthofyeastduring fermentationdependsonthemediacomposition (substrate),theinitiallevelsofpH,temperatureand dissolved oxygen (Ruiz et al., 2004). Theselectionofagoodyeaststrainhavingdesirable properties is a prerequisite for the quality wine production (Degree,1993).EnologicaltraitsofS.cerevisiaehave beendividedintotwogroups,thatis,technologicaland qualitative, and both groups have to be considered in the selectionofwineyeasts.Thetechnologicalones influencethefermentationefficiency,andthequalitative onesdeterminethechemicalcompositionandsensorial characteristics of wines. In traditional winemaking, natural fermentationofgrapejuiceiscarriedoutbyasequence ofdifferentyeastspecies.Theearlystagesofthe alcoholicfermentationaredominatedbythegrowthof non-Saccharomycesyeasts,characterizedbyalow fermentative power. Of these, Hanseniaspora (Kloeckera) andCandida(e.g.CandidastellataandCandida pulcherrima)aremorefrequentlytheprincipalyeasts observedbothinspontaneousandinoculated fermentations(HeardandFleet,1986).Thisisfollowed byappearanceofmoreethanoltolerantSaccharomyces sp.VariousstrainsofS.cerevisiaeareavailableas starterculturestosupplydistinctivesensoryattributesto wine(Cavazzaetal.,1989).Winemakersrelyonrapid fermentationsandtheyproducepredictableflavorand aromacharacteristicsinthefinishedwine.Thismakes researchongeneticsandphysiologyofwine yeast, of854Afr. J. Food Sci. paramountimportance.ThedesiredS.cerevisiae characteristicsforwinemakinginclude:Osmotolerance, relative insensitivity to high acidity, and acceptance of low oxygen concentrations.Insomecases,wineproducedwithyeastmono-cultureslacksflavorcomplexitythatmayoriginatefrom goodindigenousfermentations.Thepotentialofnon-Saccharomycesyeaststoenhancewinearomaintensity andflavorcomplexityisalsofoundtobeconsiderable. SomeofthesestrainssuchasKloeckera apiculata, Pichiafermentans,C.stellataandotherspecieshave beenstudiedfortheirinterestingorganoleptic contributions(Clementeetal.,2005;Uglianoand Henschke,2009).Moreover,thewineyeast,S. cerevisiaealsoplaysacentralroleintheproductionof volatilesulfurcompounds.Throughthesulfatereduction sequencepathway,thehydrogensulphate(HS)is formed,whichcanleadtotheformationofhydrogen sulfideandvariousmercaptancompounds,thusadding bouquet to the wine (Swiegers and Pretorius, 2007). Guava must fermentation The chief sugars of guava juice are glucose and fructose that are fermented by S. cerevisae to produce ethanol.A numberoffactorsaffectyeastfermentationperformance likethenatureofyeaststrain,fermentationtemperature, media composition, pH, substrate concentration, mode of substratefeeding,ethanolconcentrationetc(DAmore, 1992).Hence,thesefactorsmustbestudiedindetail, especiallytheinteractionsbetweenthemandtheir influenceonfermentatingmicroorganisms.Theeffectof importantfermentationparametersasfermentationof guava is described subsequenty:

Effect of temperature Temperature can affect the sensitivity of yeasts to alcohol concentration,growthrate,rateoffermentation,viability, length of lag phase, enzyme and membrane function, etc. Becauseyeaststrainsdifferinresponsetotemperature, theoptimumtemperatureforvinificationcanvarywidely. Torijaetal.(2001)observedamixedresponseto fermentationtemperature(15to35C)onmixedstrain populationofS.cerevisiae.Alcoholyieldwashigherat higher temperature while at lower temperature secondary metabolitessuchasvolatiles,esters,glyceroletc. increased(Roehr,2001;Torijaetal.,2003;Pramanik, 2003;Robinson,2006).Fermentationsconductedunder lowtemperaturesenableariseinproductionandaroma retention,whichmayfavoranimprovementinthe aromaticprofileofthewine(Torijaetal.,2003).Thisis becausefermentationsalteryeastnitrogentransportand metabolismthushamperingthecoordinationbetween carbonandnitrogenmetabolismsatlowtemperatures (Beltranetal.,2007).Ontheotherhand,high temperaturemaydisruptenzymeandmembrane functions,resultinginstuckfermentation(Seneretal., 2007).Fermentationathighertemperaturesmayhave adverseeffectonthewineinstunningtheyeastto inactivity and even "boiling off some of the flavors of the wines.Amongthedifferentreportsavailableinliteratureon guavafermentation,atemperaturerangeof25to30C hasbeenreportedfordifferentvarieties(Srivastava, 1997;Shankaretal.,2006;YuandZhang,2008;Sevda andRodrigues,2011;Pooja,2011).Sevdaand Rodrigues(2011)reportedthatforS.cervisiaeNCIM 3095,themaximumethanolproductionis7.784%(v/v) at25CandforNCIM3287,themaximumethanol production is 8.396% (v/v) at 25C.High temperature shift (Heat shock) has been used for increasing glycerol production byyeast which contributes towardssmoothmouthfeelinguavawineproduction (SevdaandRodrigues,2011).Amongthetemperature range (15 to 35C) studied by us, the results revealed the maximumethanolproductionof11.1,11and11%at 25Cinthreeguavavarietiesviz.Punjabpink,Arka amulya and Lucknow-49, respectively (Pooja, 2011). Effect of pH ThepHofthegrowthmediumisanotherimportant parameterforthesuccessfulprogressoffermentation becauseitinfluencesyeastgrowthaswellasethanol formation besides sensory quality of wine. It is known that awinewithapHoflessthan3.4presentsanotable resistancetobacterialattack.However,inawinewitha pHmorethan3.6,thedevelopmentofharmfulmicrobial floramayoccur.Ontheotherhand,fermentations conductedinexcessivelyacidicmediabecometooslow due to the low growth rate of the yeast (Ough, 1991). The pHofthemediummaychangeinresponsetometabolic activitiesofmicroorganisms.Also,duringthecourseof fermentation,nitrogensourcecansignificantlyaffectthe pHandthereforeinitialpHofthemediummustbe adjusted carefully. Infact,combinedtreatmentofthemustbypH adjustmentandsulfurdioxideadditionhasbeen consideredasanappropriatetechniquetoprevent microbialspoilageinwinefermentationbecauselowpH canimprovethepasteurizationeffectofsulfurdioxide and give the winemaker important information about how muchsulfurdioxideisneededtocontrolmicrobes effectively.Thisissignificantbecausehighsulfurdioxide contentinthemustnegativelyaffectshumanhealth (Robinson, 2003).pHhasbeenfoundtoaffectmalicacid(animportant volatilecompoundaffectstitrableacidityofwine).Ithas beenreportedthatmalolacticfermentation(MLF) rates aredirectlyproportionaltoinitialpH values and MLF is associatedwithgreaterdiacetyl, acetoin,andvolatile acidityproduction(BousbourasandKunkee,2007;Liu andGallander,1983).GuavamusthadapHrange between4.2to4.4inthreedifferentvarietiesstudiedby us(Pooja,2011)whichwassuitableforitsfermentation. ApHlevelof4.0to5.0wasfoundtobeoptimumfor guavamustfermentationinotherstudiestoo(Yuand Zhang,2008;Shankaretal.,2006;Sevdaand Rodrigues, 2011). Effect of initial sugar level Theconcentrationofinitialsugarsisanimportant parameterinthefinalethanolproductionanditssensory quality.Ithasbeenobservedthatinitialsugarlevelof juicegreatlyaffectstherateoffermentation.Useof concentrated sugar substrate is one of the ways to obtain highethanolyieldduringfermentation.However,high substrateconcentrationsareinhibitorytofermentation duetoosmoticstress(Jonesetal.,1981).Thereare reportsthathighsugarsensitivity(osmotolerance)inS. cerevisaeisduetohigherintracellularaccumulationof ethanol(StrchaianoandGoma,1983).Differentstrains presentvariablelevelsoftolerancetothisaccumulating ethanolandhencetheirrequirementofdifferentinitial sugar concentration for fermentation. In some strains, this levelisjust10to15%(Praminik,2003;Asli,2010)and thishasbeenobservedinguavafermentationalso (Cheema,1989;Srivastavaetal.,1997).However,initial sugarsashighas25to30%havealsobeenemployed successfullyforethanolicfermentation(Bertolinietal., 1991).Inguava,weoptimized25%(w/v),Sevdaand Rodriguez (2011)optimized 20 to22% (w/v) andYuand Zhang(2008)optimized20%(w/v)asinitialsugar concentration for guava wine production. Theeffectofinitialsugarconcentrationontimeof fermentationhasalsobeenobservedashighersugars tendtoprolongfermentation(Borzanietal.,1993). Higherinitialsugarsalsopossessbetterretentionof ascorbicacid,increaseinconcentrationoftotalesters and phenols thus improving the wine quality (Attri, 2009). Effect of inoculum size Thestandardizationofinoculumsizeisimportantas sugarconsumptionisabalancebetweenbiomass development and ethanol production and a high inoculum sizewillthusbeacompromiseonamountofethanol produced.Wehaveobservedthatethanolproduction increaseswithincreaseininoculumconcentrationupto 9%(v/v)anddecreasessignificantlybeyondinoculum levelof9%(v/v)incaseofthreevarietiesofguavaviz; Punjab pink, Arka amulya and Lucknow-49 (Pooja, 2011). SimilartrendshavealsobeenreportedbySinghand Kaur (2009) where they observed 10% (v/v) as optimized inoculum level for litchi wine production.Kocher and Pooja855 Inguava,Srivastavaetal.(1997)reportedthat10% inoculum size added in non chaptalized guava pulp led to the production of 5.8% ethanol (w/v) by S. cerevisae.An optimizedinoculumlevelof10%v/vforalcoholic fermentation of jamun, plum, apple, pear juice, guava and 7.5% inoculum size for kinnow wine production has been observedinotherresearchreports(Tewarietal.,1987; Cheema, 1989; Panesar et al., 2009). Effect of nitrogen and phosphorus sources Theintrinsicimportanceofassimilablenitrogentoyeast growth and metabolism is a well known recognized fact in winemaking.Factorssuchasthejuicecompositionand thekindoftheyeaststraincanaffecttheassimilationof thenitrogenouscompounds(Colombietal.,2007; Manginotetal.,1998)asthemetabolismofaminoacids can affect the efficiency of the alcoholic fermentation and thequalityoftheproduct(Berthelsetal.,2004). Imbalances and in particular deficiencies in the supplyof assimiablenitrogencompoundshasremainedasthe mostcommoncausesofstuck/sluggishfermentation. Presence of sufficient amount of nitrogen in the fruit juice isreportedtoenhancetheyeastgrowthandsugar catabolicrate.However,additionofexcessNandP sourcecaninhibitthefermentationprocess.Hence addition of N and P sources in an optimum quantity is an important fermentation parameter. Further,fermentationprocessnotonlydependsupon thequantityofNandPadded,butalsoonthequalityof thesesupplements.Variousnitrogenandphosphorus sourceslikeammoniumsulphate,diammonium phosphate,potassiumdihyrogenphosphate,di potassiumhygrogenphosphate,aminoacids,etc.have been used to carry out fermentation efficiently and rapidly in different fruit wine fermentations (Ough, 1991; Shankar et al., 2006; Soni et al., 2009; Asli, 2010; Pooja, 2011).Ithasbeenfoundthatdi-ammoniumhydrogenortho phosphate(DAHP)supplementationimprovesthewine colour,totalacids,bouquet,taste,aromaandoverall sensoryquality.Shankaretal.(2006)reportedthatwine producedfromGuavapulp(1:4dilutionwithwater)with 0.1%DAHPsupplementionwasbetterinitssensory attributesandhadthehighpercentageethanolthanthe non supplemented one. Patil and Patil (2006) had similar observations with pineapple wine fermentation. The effect ofDAHPsupplementationattherateof0.3%(w/v) producedsignificantlyhigherethanolpercentageof 13.60.2withafermentationefficiencyof93.80.8%in threedifferentvarietiesofGuava(Pooja,2011).A schemeforpreparationofguavawineispresentedin Figure 1. POST FERMENTATIVE TREATMENTS Consumeracceptabilityisthe final goal of wine 856Afr. J. Food Sci. Figure 1. Scheme for preparation of Guava wine. development. The crude wine obtained after fermentation containsyeast,proteinhazes,residualsugarsanddoes nothavecompletequality(sensory)attributes.Hence,it requirespostfermentativetreatments(finishing)tomake a wine potable. In fact, there are five goals of "finishing" a wine:clarity,stability,compositionaladjustment,possible blendingandpackaging.Itisimportant,especiallyin white wines like guavawine, that thewine at the point of consumption should not be cloudy or contain any haze or precipitate.Itisalsoimportanttopreventunwanted microbialgrowthfromoccurringinthewineafterthe primary fermentation is complete as exposure to air (after CO2blankethasdissipated)willaffecttheflavorand aroma profile (Robinson, 2003). Racking Racking is theprocess of siphoning thewineinto anew, cleanbarrel.Rackingallowsclarificationandaidsin stabilization. Wine that is allowed to age on the lees often develops"off-tastes".Arackinghoseortubingisused and canbeattachedtoa racking coneto make this task easier.Therackingprocessisrepeatedseveraltimes during the aging ofwine. Repeated racking produces the clarityrequiredinwine,especiallyifitisagedinabarrel (Robinson,2003).Besidesclarification,rackingalso provides suitable conditions for oxygen to dissolveinthe wine,ataratevaryingfrom2.5to5mg/L.Oxygen eliminatescertainunpleasantreductionodors(H2S),as wellasiron(ferriccasse)andisalsoresponsiblefor intensifyingcolorofwine(Ribereau-Gayonetal.,2000). Weobservedthatduringthestorageperiod(upto90 days)ofguavawine,variousparameters(pH,ascorbic acid,totalphenols,percentageethanol)decreased significantly.Inspiteofdifferencesininitialethanollevels the final percentage ethanol (at 90 days) was constant at 12.6to12.8whicharereasonablygoodforwine(Pooja, 2011).YuandZhang(2008)reportedfiltrationand pasteurizationofyoungguavawinetoprepareaclear guava wine. Effect of fining agents Fining is the non-mechanical removal of unwanted and/or unstable elements in juice or wine by the addition of inert andadsorptivesubstances.Itgenerallyinvolvesthe formation of an insoluble deposit which is separated from theliquidbyeither filtration or racking. Interactions mayKocher and Pooja857 Table 1. Effect of storage time on microbiological and physicochemical properties of wine var. Punjab pink, Arka amulya and Lucknow-49.

Storage time (days) Parameters Percentage ethanol(v/v)pHAscorbic acid (mg/100ml)Total phenols (mg/100ml)Total yeastcount (cfu/ml) Punjab pink Arka amulya L-49 Punjab pink Arka amulya L-49 Punjab pink Arka amulya L-49 Punjab pink Arka amulya L-49 Punjab pink Arka amulya L-49 013.813.513.53.43.63.583.676.091.2337.0268.0246.07.3X1066.2X1067.0 X 106 1513.713.313.53.43.63.580.374.388.7326.0257.0232.02.1X1011.5X1011.1 X101 3013.513.213.33.43.53.477.670.686.0318.0242.0229.00.3X1010.6X1010.4X101 4513.513.213.13.33.53.475.268.484.1315.0231.0201.00.00.00.0 6013.313.012.83.23.53.471.967.280.3302.0229.0194.00.00.00.0 7512.913.012.73.33.43.467.065.178.9287.0215.0189.00.00.00.0 9012.812.812.63.33.43.463.064.276.0281.0211.0180.00.00.00.0 CD (5%)0.291NS0.7540.984- Residual sugars in all the varieties were not detected even at 0 day of storage. include electrostatic charges, hydrogen bonds, ion exchangeandhydrophobicreactions.Fining agents are used to achieve clarity and to improve color,flavorandphysicalstabilitye.g.,Earths (bentonite),proteins(gelatin,isinglass,casein, albumen),polysaccharides(agars),carbons, syntheticpolymers(PVPP),silicondioxide (kieselsol)andothers(includingchelatorsand enzymes). Certaingelatinscansignificantlyreduceacetic acidbacteriaandyeastpopulations,compared with samples thathavebeen racked butnot fined (MuratandDumeau,2003).Clarificationofapple juicebyflocculationandprecipitationwith bentoniteandgelatinwasdeterminedbyturbidity andzeta-potentialwhichwastreatedwith Polyvinylpolypyrrolidone(PVPP)toremovetotal polyphenol.Resultsindicatedthatriskofhazeby freegelatininjuicerequiredatleasttentimes moregelatinthantheoptimumdosagefor clarification (Benitez and Lozano, 2007). Theeffectsofdifferentfiningagents,usedat differentconcentrations,ontheantioxidantstatus offinedwineswerestudiedbyYildirim(2011). Theresultsdemonstratedthattheuseofa combinationofgelatinandKieselsolledtothe highesttotalphenolvalue(3,491mg/LGAE)and antioxidantactivities(29%)amongthetested finingagents.Theresultsofthegroupingof analyzed parameters in n-dimensional space, with differentfiningagentsatdifferentconcentrations, demonstratedtheimportanceofalow concentrationoffiningagentsforhighantioxidant activityandtotalphenols.Inliterature,however wedidnotcameacrossanyreferenceoffining the guava wine. Ageing Theageingofwineanditsabilitytopotentially improvewinequalityforits consumption, is one ofthemostimportantstepafterwineproduction (Robinson,2006).Theratioofsugars,acidsand phenolicstowaterisakeydeterminationofhow wellawinecanage.Highertemperatures acceleratetheagingprocessdramatically(e.g, storingwineat59degreesages50%fasterthan 55degrees).Fasteragingincreasestherateof undesirablechemicalreactionswhichcan produce compounds with foul odors and off tastes (Robinson,2006).Ageinginwoodenbarrels improvesthewinequalitybyaddingdesirable components including ethyl acetate, phenolics etc. and leads to reduction in undesirable components suchasn-proponal,n-butanol,iso-butanol, isoamylalcohols(Sonietal.,2009).Sonietal. (2009)alsoreportedthatstorageofamlawinein oakwoodenbarrelsforamonthimprovesthe quality and sensory attributes than the wine stored in glass bottles. We stored our guava at 15C and recorded significant decrease in phenols, ascorbic acid,ethanolandviablecount. The final wine at858Afr. J. Food Sci. theendof3monthshad12.60.2%ethanol(v/v), 67.737.18ascorbicacid(mg/100ml),and224.051.7 totalphenols(mg/100ml)andwasfreeofviableyeast cells (Table 1). Sensory evaluation Thesensoryanalysisofwineisanimportantparameter in determining the quality of wines. It revolves around the taste, feel, aroma and bouquet of the aged wine. A numberofmethodsintheformofhedonicscalesand analyticaltechniqueslikeGCMShavebeendeveloped (Amerine and Roessler 1976; Reynolds, 2010). So much istheimportanceofsensoryevaluationthatcapturing consumersmindandattitudestowardswinesisa flourishingbusiness(Lesschaeve,2007).Shankaretal. (2006)highlightedtheincreaseinaromaandflavorof guavawinewithsupplementationofNandPinthe must.Wealsofoundthatguavawinefromthree varieties that is, Punjab pink, Arka amulya and Lucknow-49hadenhancedtaste,aromaandflavorwithageingof three months (Pooja, 2011). CONCLUSIONS AND FUTURE PERSPECTIVES Wineisoneofthefunctionalfermentedfoodshaving manyhealthbenefitslikeanti-ageingeffects, improvementoflungfunction(fromantioxidantsinwhite wine),reductionincoronaryheartdisease,development ofhealthierbloodvesselsandreductioninulcer-causing bacteria.Manywinesaremadefromfruitshaving medicinalvalue(Tapselletal.,2006).InIndia,wine industryisgrapebasedandisstillinitsinfancy(Joshi andAttri,2005)withwineriesrestrictedtoMaharashtra. Out of the total 60 wineries in the country, 57 are located inMaharashtra,2inKaranatakaand1inGoa(Patil, 2008).AsfarasPunjabisconcerned,inspiteof61,618 haunderfruitsandaproductionof10,55,408MT annuallyproductionwithguavacontributing1.5lakhMT (Gill et al., 2009), no winery has been set up here yet. 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