ORIGINAL ARTICLE
Production of Star Fruit Alcoholic Fermented Beverage
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´via de Paula Valim 1•Elizama Aguiar-Oliveira 2•Eliana Setsuko Kamimura 3•
study是什么意思
Vanessa Dias Alves 1•Rafael Rende Maldonado 1,4
Received:19January 2016/Accepted:18May 2016/Published online:28May 2016ÓAssociation of Microbiologists of India 2016
Abstract Star fruit (Averrhoa carambola )is a nutritious tropical fruit.The aim of this study was to evaluate the production of a star fruit alcoholic fermented beverage utilizing a lyophilized commercial yeast (Saccharomyces cerevisiae ).The study was conducted utilizing a 23central composite design and the best conditions for the production were:initial soluble solids between 23.8and 25°Brix (g 100g -1),initial pH between 4.8and 5.0and initial con-centration of yeast between 1.6and 2.5g L -1.The conditions yielded a fermented drink with an alcohol content of 11.15°GL (L 100L -1),pH of 4.13–4.22,final yeast concentration of 89g L -1and fermented yield from 82to 94%.The fermented drink also prented low levels of total and volatile acidities.
Keywords Star fruit ÁFruit wine ÁFactorial design ÁSaccharomyces cerevisiae
Introduction
The alcoholic fermentation of fruit can be ud for the production of alcoholic drinks and it is commonly realized by yeast such as the Saccharomyces cerevisiae [1,2].In such process occurs the production of ethanol and carbon dioxide,which are obtained by the anaerobic conversion of the sugars naturally contained in the fruit or added to it.According to the Brazilian legislation [3],wine is a drink with alcohol content from 4to 14°GL (L 100L -1)at 20°C,produced from the alcoholic fermentation of healthy,ripe and fresh grapes.The term ‘‘fruit wine’’is applied to alcoholic fermented drinks produced from fruits other than grapes.Any fruit with reasonable amounts of fer-mentable sugars can be utilized as must for the production of wine.The usage of different fruits may lead to the obtain-ment of drinks with different flavors.Therefore,many exotic fruits have been utilized in the production of wine.
Carambola or star fruit (Averrhoa carambola )is a tropical fruit originally from Indonesia and India,being very popular in South-eastern Asia,South Pacific and some regions of Eastern Asia.The carambola tree is grown in other countries out from Asia,such as Colombia,Guiana,Dominican Republic,Brazil and the USA [4].The star fruit chemical characteristics depend on climatic factors,the type of soil utilized for cultivation,the fruit ripeness level,etc.Almeida et al.[5]characterized ripe star fruit from north-eastern Brazil and obtained average values of soluble solids and pH of 8.0°Brix and
3.7,respectively.Star fruit is rich in vitamins,oxalic acid,polyphenols,dietary fiber,volatile compounds,etc.Such traits allow innumerable usages for the fruit,as well as providing benefits for the health of the consumers [6,7].
The aim of this study was to evaluate the production of a star fruit alcoholic fermented drink incorporating the fruit
Electronic supplementary material The online version of this article (doi:10.1007/s12088-016-0601-9)contains supplementary material,which is available to authorized urs.&Rafael Rende Maldonado
br
1
Municipal College Professor Franco Montoro (CMPFM),R.dos ,Mogi Guac ¸u,SP 13.843-971,Brazil 2
Multidisciplinary Institute on Health (IMS),Federal
University of Bahia (UFBA),campus Anı
´sio Teixeira (CAT),R.Rio de Contas,58,Vito
´ria da Conquista,Bahia 45.029-094,Brazil
3
Faculty of Zootecnic and Food Engineering (FZEA),
University of Sa
˜o Paulo (USP),Av.Duque de Caxias,225,13.635-900Pirassununga,Sa
˜o Paulo,Brazil 4
Technical College of Campinas (COTUCA),University of
Campinas (UNICAMP),R.Jorge Figueiredo Corre
ˆa,735,Pq.Taquaral,Campinas,Sa
˜o Paulo 13.087-261,Brazil Indian J Microbiol (Oct–Dec 2016)56(4):476–481DOI 10.1007/s12088-016-0601-9
五年级下册英语电子书
traits in the wine.The effects of the initial concentration of visiae),initial pH and initial sugar concen-tration were evaluated utilizing a23central composite design whereby the fermentation kinetics data and the physico-chemical characteristics of the product were analyzed.
Materials and Methods
Preparation of the Star Fruit Pulp and Must
For the star fruit pulp preparation was lected ripe fruits with good appearance,smell and texture.They were bought at a local supermarket in the region of Campinas-SP,Brazil, in March2014.The star fruits were cleaned,cut and blitzed in a blender until a pulp consistency was achieved.There-after,the pulp wasfiltered with be means of a cotton cloth, in order to remove the insoluble solids and then pasteurized at80°C for5min.The pasteurized pulp was transferred to plasticflasks which were clod and left cooling to room temperature and then frozen and stored at-18°C to the moment they were utilized on the must preparation.
In order to prepare the must for the fermentation,the star fruit pulp was thawed at room temperature(*25°C)and mixed with saccharo to adjust the concentration of sol-uble solids indicated on Table1(varying between19and 25°Brix).Thereafter,calcium carbonate was added to eac
h test to adjust the pH,also regarding the values indicated on Table1(ranging between4.0and5.0).
Alcoholic Fermentationironic
The must obtained was then utilized for the alcoholic fer-mentation.A lyophilized commercial visiae (Fermentais Lessaffe GroupÒ)was utilized.The lyophi-lized yeast was rehydrated with a small portion of must and thereafter added to the total must volume(1.0L).All fer-mentations were conducted at room temperature(*25°C), without agitation in glassflasks covered with plasticfilm, in which small orifices were made to facilitate the elimi-nation of the carbon dioxide produced during the fermen-tation process.Each fermentation lasted9days and the soluble solids consumption of the must,as well as its pH, were measured daily.By the end of the fermentation,the alcoholic content and the yeast concentration were also measured.The total,fixed and volatile acidities were also analyzed for each wine obtained.
Factorial Design
The study of the star fruit alcoholic fermentation by S. cerevisiae was conducted utilizing a central composite rotatable design with23factorial points?6star points?3 central points,totaling17tests[8]with independent vari-ables:soluble solids concentration(SS)(19a25°Brix), initial pH(4.0–5.0)and initial ye
ast concentration(IY)(1.0 a4.0g L).The effects of the three variables on the fer-mentation process respons(alcoholic content(AC),final pH,volumetric fermented yield(VFY)andfinal yeast concentration(FY))as well as on thefinal product(total,fixed and volatile acidities)were evaluated.The results obtained were analyzed by means of the software Statistica 8.0(Statsoft)and the factorial design matrix is shown on Table1.
Analytical Methods
The pH was measured directly with a bench pHmeter(Bel EngineeringÒ,W3B model).The soluble solids (S)(°Brix=g100g-1)were determined with a portable refractometer(InstrutempÒ,model ITREF25). From the S values,and knowing the stoichiometry ratio of sugar consumption/ethanol production(1:4mol of sac-charo per mol of ethanol)it was possible to estimate the alcohol content as the function of the fermentation time.At the end of each fermentation a sample was collected to measure the alcohol content(°GL=L100L-1)in an Alcolyzer equipment(Anton PaarÒ).The conversion fac-tors:fermentable sugars into biomass(Y X/S=dX/-dS), fermentable sugar into alcohol(Y P/S=dP/-dS)and bio-mass into alcohol(Y P/X=dP/dX)were also calculated considering(-dS:g L-1)the fermentable sugar con-sumption,(dX:g L-1)the cell growth,and(dP:g L-1)the alcohol production.Thefixed and total acidities(meq L-1) were measured using titrimetric methods according
to Adolfo Lutz Institute[9];the volatile acidity was calcu-lated by the difference between total andfixed acidities. Results and Discussion
耆寿耇老The respons obtained from the central composite rotat-able design applied for the production of star fruit alcoholic fermented visiae can be obrved in Table1. From the results,the analysis of variance(ANOVA) (Online Resource1)was performed for each respon and cond order models were evaluated to explain the process.
Considering the ANOVA results for the alcohol content (Online Resource1),it was possible to obtain a significant and predictive codified model with94%confidence (p=0.06),reprented by Eq.1,where OH is the alcohol content(°GL),S is the initial soluble solids concentration (°Brix),P is the initial pH and Y is the initial yeast con-centration(g L-1).The resulting surface respons and contour lines can be obrved on the Online Resource2.
The analysis of thisfigure shows that the best results for the AC,after9days of fermentation,were obtained from the highest soluble solids concentrations,the highest pH and the initial yeast concentration levels between-1 (1.6g L-1)and?1(3.4g L-1).The conditions,pre-dicted by the model,can be verified in Table1,wherein the highest alcohol content were obtained in trials4,8,10and
12,which resulted in values from10.45to11.15°GL.The results obtained in this study were better than others cited in the literature,which also ud star fruit as a substrate for alcoholic fermentation.Napahde et al.[10]obtained only 0.2°GL of alcohol after21days of fermentation,while Sibounnavong et al.[11]obtained8.3°GL(average)after 2weeks.In other study,Bridgebassie and Badrie obtained a similar alcohol content(from10.25to11.50°GL)after 4weeks of fermentation using star fruit must pretreated with different concentrations of pectola and using dif-ferent yeast strains[12].
The best conditions(trials4,8,10and12)were lected for the analysis of their consumption of soluble solids and the ethanol production during the fermentation time.The obtained kinetic profiles can be en in Fig.1.The con-sumption of soluble solids and the ethanol production showed similar profiles for the lected assays,indicating no significant difference among the chon conditions.It was also possible to e that after5days of fermentation there was a trend for stabilization(Fig.1),with low sugar consumption and little ethanol production.
OH¼10:01þ0:61SðÞÀ0:05S2
ÀÁ
þ0:34PðÞÀ0:05S2
ÀÁþ0:34PðÞÀ0:05P2
三月英文简写ÀÁ
þ0:02YðÞÀ0:42Y2
ÀÁþ0:44SðÞPðÞÀ0:26SðÞYðÞþ0:22PðÞYðÞð1ÞRegarding to thefinal pH of the star fruit alcoholic fermented,it was not possible to obtain a predictive model (p\0.10)to reprent the process(Online Resource1).By obrving Table1,however,it was possible to notice that the averagefinal pH value,considering the17trials,was 4.05and the initial value was4.50,this reduction is a good indicator for the fermentation process becau it indicates that most of the substrate was ud for ethanol production and cellular growth and there was little formation of acid. Excessive production of acids may indicate microbial contamination by acetic bacteria,excess oxygen in the fermentation medium or excessive fermentation time. Bridgebassie and Badrie[12]obtained a lowerfinal pH of about3.1in star fruit wine,but the authors applied a pretreatment in the must using1%of citric acid before the fermentation.
Similarly,for the volumetric fermented yield,the cor-relation with the independent variables was very low (R2=0.24)and thus,none of the studied variables had a
Table1Kinetic parameters for star fruit alcoholic fermentation at room temperature after9days of fermentation
星期一至星期日的英语单词Runs SS(°Brix)pH Yeast(g L-1)Alcohol(°GL)Final pH Yield(%v/v)Final yeast(g L-1)
120.2(-1) 4.2(-1) 1.6(-1)9.06 3.7485.4783.94
223.8(?1) 4.2(-1) 1.6(-1)9.76 3.7785.0054.58
320.2(-1) 4.8(?1) 1.6(-1)8.36 4.2390.0065.22
423.8(?1) 4.8(?1) 1.6(-1)11.15 4.2294.0089.66
520.2(-1) 4.2(-1) 3.4(?1)8.36 3.8285.5088.14
623.8(?1) 4.2(-1) 3.4(?1)8.36 3.8790.1074.54
720.2(-1) 4.8(?1) 3.4(?1)9.06 4.2491.5063.85
823.8(?1) 4.8(?1) 3.4(?1)10.45 4.2190.8070.74
919.0(-1.68) 4.5(0) 2.5(0)9.06 4.1178.6091.73
1025.0(?1.68) 4.5(0) 2.5(0)11.15 4.1381.7489.84
1122.0(0) 4.0(-1.68) 2.5(0)9.76 3.9483.33131.81
1222.0(0) 5.0(?1.68) 2.5(0)10.45 4.1975.76167.08
1322.0(0) 4.5(0) 1.0(-1.68)8.36 4.0583.33144.86
1422.0(0) 4.5(0) 4.0(?1.68)9.76 4.0082.69121.88
1522.0(0) 4.5(0) 2.5(0)9.76 4.3587.78190.80
1622.0(0) 4.5(0) 2.5(0)10.45 4.0075.30213.07
1722.0(0) 4.5(0) 2.5(0)9.76 4.1077.69190.89
Central composite rotatable design23?6star points?3central points for the star fruit alcoholic ferment
ation visiae after9days at room temperature(*25°C)without agitation.Independent variables are:the soluble solid concentration(S)(°Brix=g100g-1),initial pH and initial yeast concentration(Yeast)(gL-1).Respon variables are:thefinal alcohol content(Alcohol)(°GL=L100L-1),final pH,volumetric fermentation yield(Yield)(%v/v)andfinal yeast concentration(Final Yeast)(g L-1)
Codified values are prented in parenthesis
significant influence on this respon (Online Resource 1).Considering the 17trials (Table 1),the average volumetric fermented yield was 84.6±5.6%(L 100L -1).Volume variations in similar process are common and can occur due to the consumption of must to produce gas (carbon dioxide)during the fermentation.
Regarding to cell ,the final yeast concen-tration,the ANOVA (Online Resource 1)proved that it was possible to obtain a statistically significant and predictive model at 93%of confidence (p =0.07).The complete codified model obtained is reprented by Eq.2where FY is the final yeast concentration (g L -1),S is the soluble solid concentration (°Brix),P is the initial pH,Y is the initial yeast concentration (g L -1).The resulting surface respon and the contour curves can be obrved on the Online Resource 3,in which it appears that the final con-centration of yeast (FY )was the highest in the conditions clo to the central points.However,when it comes to
alcoholic fermentation process,the best condition for cell growth is not necessarily the best condition for ethanol production.Reddy and Reddy [13]evaluated the effect of different parameters over visiae growth in mango must and it was obrved a great influence of temperature;at 25and at 30°C they obtained the highest cell populations within 6and 8days of fermentation,respectively,with an ethanol production 70%smaller at 25°C than at 30°C (41.3g L -1day -1).
FY ¼201:32À1:08S ðÞÀ48:35S 2ÀÁþ3:48P ðÞÀ27:56P 2
ÀÁ
À2:55Y ðÞÀ33:25Y 2ÀÁ
þ9:28S ðÞP ðÞÀ0:22S ðÞY ðÞ
À5:56P ðÞY ðÞ
ð2Þ
The conversion factors were also calculated for each trial (Online Resource 4).The ANOVA was also obtained (data not prented),but it was not possible to obtain cond order models to explain each
chine foodof the conversion factors with a reasonable level of confidence.The results (Online Resource 4)indicate that the highest values of Y X/S and Y P/S were obtained in trial 12,however,the highest value of Y P/X was obtained in trial 2.It confirms the pre-viously mentioned suggestion that a greater microbial growth does not necessarily result in a greater ethanol production.Confronting the three conversion factors and the two models obtained (for OH and FY),the best con-ditions for the production of star fruit alcoholic fermented (aiming the highest alcohol content)were tho in which conditions where the substrate was better ud for con-version into product than cell growth.The conditions correspond to trials 4and 10which prented the highest initial concentrations of soluble solids (23.8–25.0°Brix),the highest initial pH (4.8–5.0)and initial concentrations of yeast in the intermediate level (1.6–2.5g L -1).
Regarding to the acidity levels (Online Resource 4),the obtained values were also evaluated by ANOVA (data not prented)but none of the three respons there were sig-nificantly correlations with the independent variables of the process.This means that under all conditions evaluated the resulting acidity values were very similar to each other,a fact that is in agreement with the analysis for the final pH of the fermentation,which had the same behavior.The composition of the acidity of star fruit alcoholic fermented (Online Resource 4)suggests that most of it is related to the fixed acidit
y (89%)and a small part (11%)to the volatile acidity.This is an excellent result,since excessive volatile acidity may indicate microbial contamination or excess of oxygen in both production and storage of fermented drink.Paul and Sahu [14]obtained star fruit alcoholic fermented with a titratable acidity of around 119meq L -1and,despite of all the other similar parameters obtained,this value is almost 2.5times higher than the average value obtained in our study (48.8meq L -1
).
Fig.1a Soluble solids (°Brix =g 100g -1)consumption and b production of alcohol (°GL =L 100L -1)in star fruit alcoholic fermentation visiae as a function of the fermentation time.The conditions for the concentration of soluble solids (°Brix)/pH/concentration of yeast (g L -1)were:23.8/4.8/1.6(trial 4);23.8/4.8/3.4(trial 8);25.0/4.5/2.5(trial 10);and 22.0/5.0/2.5(trial 12).The experiments were conducted at room temperature (*25°C)without agitation,for 9days.Lines were ud to connect the points and guide the eyes
Brazilian law,for instance,specifies some quality stan-dards for grape wines[3],but it does not specify any stan-dard for wines from other fruits.Comparing with the current legislation,star fruit wine obtained showed an average total acidity of48.8±7.7meq L-1similar to the minimum established by Brazilian law(55meq L-1);however vola-tile acidity was much lower,5.4±2.0meq L-1,than the maximum established(20meq L-1)indicating low levels of acetic acid,or low oxidation of ethanol.The volatile and total acidities of jabuticaba(Myrciaria jaboticaba)wines determined by da Silvaet al.[15],were higher than tho obtained in our study,with values above185and 17meq L-1,respectively.
Other study conducted for the production of watermelon (Citrullus lanatus var.Lanatus)alcoholic ferm
ented[16] led to similar results to tho obtained with star fruit fer-mented for:pH(4.1),final concentration of SS(6.6°Brix), alcohol content(10°GL),fermented yield(94%)and Y P/S (0.67).However thefinal biomass concentration (20g L-1)and Y X/S(0.14)were lower and the fermenta-tion time(48h)was reduced comparing to tho obtained with star fruit wine.The results indicate that the alco-holic fermentation with star fruit was predominantly anaerobic,with good ethanol production.A wine made from jackfruit(Artocarpus heterophyllus)[17]stored for 11months prented a higher alcohol content(13°GL)and a higher total acidity(100meq L-1),but the volatile acidity was very similar to the star fruit wine(6meq L-1).
Using a similar initial concentration of yeast (1.65g L-1),Andrade et al.[18].had a slightly more acidic wine(pH=3.51)from strawberry(Fragaria ana-nassa)and similar alcohol content(9.62°GL)after 30days of fermentation,with soluble solids consumption varying from27to9°Brix.For this product,the authors obrved that the consumption of soluble solids were more inten in thefirst10days.Tamarind(Tamarindus indica) and soursop(Annona muricata)were also objects of study [19]for the production of alcoholic drinks,resulting in alcohol levels of8.1and6.3°GL,respectively.Star fruit was applied by Paul and Sahu[14]and it was obtained an alcoholic fermented drink with similar alcohol content of 12.15°GL,however the authors obtained hi
gher titrable acidity(0.76%w/w),lower pH(3.94),lower total soluble solid concentration(4.6°Brix)and they ud an inoculum step which added more time to the process. Conclusion
At the initial conditions of:23–25°Brix;pH=4.8–5.0and 1.6–2.5g L-1of yeast concentration,it was possible to obtain a fermented drink,from star fruit,with:an alcohol content of11.15°GL,pH from4.13to4.22,afinal yeast concentration of89g L-1,volumetric fermentation yield from82to94%(v/v),total acidity from42to52meq L-1, a volatile acidity of5meq L-1andfixed acidities of 37–47meq L-1.The average conversion factors were:Y X/ S
=0.79;Y P/S=0.67and Y P/X=1.02.The fermentation kinetics showed that after5days of fermentation(*25°C), the process reached thefinal stage.The obtained drink had similar characteristics to drinks made from other fruits mentioned in the literature and the differences among their parameters are basically due to the differences in the pro-cess and in the compositions of raw materials ud. Acknowledgments Authors are grateful to the Municipal College Professor Franco Montoro(Mogi Guac¸u,Sa˜o Paulo,Brazil)where all the experiments were conducted and also to the Coordenac¸a˜o de Aperfeic¸oamento de Pessoal de Nı´vel Superior(CAPES,Brazil)for theirfinancial support.
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