iersrgltslbln eurmns sdachlzdwine dealcoholized and Tobacco As Alcohol, requirements. of labeling Bureau regulates the while Firearms alcohol, v/v within 7% 24% between defined and contain as must re- Act, wine Administration States, Alcohol and United Federal the the wine In regulating wine. alcohol authorities duced government arising complexity different legislative have from con- countries be contain Some to wine. still concentration a may ethanol sidered specified but minimum levels the than alcohol less modify postfer- to any process undergone have mentation not con- may alcohol products low These naturally centrations. of consequently, levels low and occurring sugars, naturally fermentable with grapes harvested techniques early production using wine from (ANZFA ethanol arises confusion v/v Further 0.5% 2002a). than less “Non-intoxicating” contains and juice beverage ethanol the grape v/v implies 1.15% fermented than from less derived Alcohol” contains beverages “Low that to term applied The as be Product.” branded may Wine market be Alcohol could Australian product “Reduced the the a however, for permitted; destined be product not may a Alcohol “Reduced describe term the to of grape use Wine” may the amount Thus, Fermented this definition. than this than 2002b). fit less greater not concentration (ANZFA alcohol contain an (alcohol) with must products ethanol in wine example, v/v as For 8% labeled countries. different products for Australia, vary- and applicable of wine concentrations quantity of ethanol ing Definitions minimum complexities. specify legal products wine with fraught is ages ute erdcinwtotpriso sprohibited is permission without reproduction Further 10.1111/j.1750-3841.2011.02448.x doi: author to inquiries Direct Australia. Charles Sciences, 2678, Wine NSW [email protected]). and (E-mail: Wagga, Agricultural Schmidtke, Schmidtke of Wagga authors School and Univ., Centre, with Sturt Industry are Grape Agboola and and Wine Blackman, Natl. with and are Schmidtke Authors Blackman 12/9/2011. Accepted 5/7/2011, Submitted 20110811 MS Introduction Agboola O. Samson and Blackman, W. John Schmidtke, M. Leigh Wines Alcoholic Reduced for Technologies Production C h auatr n aeo eue looi teghbever- strength alcoholic reduced of sale and manufacture The 01Isiueo odTechnologists Food of Institute 2011 cerevisiae h nomto rsne nti eiwwl lo ieaest sesterltv ehia eiso aho the of reducing each for of techniques merits winemaking technical novel relative of the implementation assess regarding to wine. in decisions winemakers concentration allow make ethanol generally will and production. review is alcohol described removal for this expression technologies alcohol in gene modified presented the for or and downregulated distillation information processes with production strains The thermal yeast nonthermal specific on utilize the of attention selection that of focused or has development acceptance techniques consumers and from by arise perception technologies perception naturally manufactured negative would consumer This postfermentation However, wines unfavorable. challenges. than or techniques. concentrations marketing of ethanol production pre- and quality lower wine technical with upon sensory and wines of focus of fermentation number production that normal a for strategies through patented poses production and style, described wine wine been specific and have a solutions for engineering acceptable Several than greater levels alcohol Keywords: Abstract: pnigcn,winemaking cone, spinning , loo euto,gntcegneig lcs xds,omtcdsilto,rvreosmosis, reverse distillation, osmotic oxidase, glucose engineering, genetic reduction, alcohol h rdcinadsl fachlrdcdwns n h oeigo tao ocnrto nwnswith wines in concentration ethanol of lowering the and wines, alcohol-reduced of sale and production The R ietedadlwrachlcnupinrtsaeassociated are rates al- world- consumption healthier alcohol a for lower is demand and consumer consumption trend a alcohol wide satisfy Decreasing to 15 products. order last coholic in the y over developed 20 been to have beverages strength coholic government and manufacturer. that bodies with for regulatory appropriate wines the organizations to from of advised opinion strongly range expert are a seek concentrations develop ethanol ex- normal to and than wish manufactured lower are who alcohol Producers reduced ported. zones with trading goods economic which production or in wine countries or specific labeling with fer- associated of laws in discussion concentrations a guide ethanol or beverages, a reducing mented for be techniques employed production to be various intended the can not of that otherwise is or article legalities the This of to definitions exist. legal may produc- specific terms that wine these acknowledged of is context it the although in tion, interchangeably low used and are dealcoholized, reduced, alcohol alcohol terms Through- notable. the review, are this upon styles out wine based specific exceptions individual of and production for zones, historical laws defi- trading labeling economic Such specific or permissible. the countries be to Oort may according (van products vary regions nitions food or or pro- products countries as wine some permitted in Canal-Llaub be wines and not the for for may aids enzymes sugars cessing (GOX) fermentable oxidase of glucose con- reduction of alcohol the use reducing may The of technique confusion centration. the Further to unsuitable. according arise term also this render could sugar) rdcsuulycnanls hn7 / tao,lbln pro- labeling ethanol, v/v 7% than less contain usually products rmgaejieta a enfretdt rns ( dryness to fermented been has or that distillation juice prepared wines grape thermal from from alcohol of some of reduction use for for the processes suitable however, membrane be units; may keeping Product” term stock the the Wine of by use Fermented administered The act 2005). “Partially (Anon federal Administration separate Drug and a Food by covered are visions rdcintcnqe o auatrn o rrdcdal- reduced or low manufacturing for techniques Production o.7,N.1 2012 1, Nr. 71, Vol. rs20) u hi s ntemnfcueof manufacture the in use their but 2002), eres ` r ora fFo Science Food of Journal Saccharomyces < . g/L 2.0 R25

R: Concise Reviews in Food Science -lactone spon- δ As gluconic acid is GOX (EC 1.1.3.4) is an aerobic gly- The most efficient conversion of glu- -D-glucose to D-glucono-1,5-lactone (D-gluconic acid β Effect of pH on GOX. Biochemical principle. Treatment of grape juice with GOX. -lactone). This reaction requires the presence of molecular oxy- δ gen, and a flavinelectron adenine donation dinucleotide to cofactorcatalase, form to is participate hydrogen frequently in peroxide.to present A degrade in 2nd the commercial unwanted enzyme, peroxideduring GOX that the preparations oxidation is of formed substrate. D-gluconic as acid a by-product juice range from lessoxidation than is 4% dependent todissolved 40%. upon oxygen The concentration, enzyme processing efficiency time, concentration, of and(Pickering temperature glucose juice and others pH, 1998; Pickering 2000). cose to gluconic acid in grape juice is reported to occur in a pH coprotein with dehydrogenase activitytion of that catalyzes the oxida- taneously hydrates to formfor gluconic these acid. reactions is The shown biochemical in basis Figure 1. not fermented by yeasts,achieved a in decrease wines in prepared alcoholGOX from production in GOX-treated can juice. be wine Thefrom was use dissolved of first oxygen described (McLeodThe for and treatment rapid Ough of 1970; protectionconcentration grape Ough of and juice 1975). wine consequently with alcoholwas explored GOX levels and in for further refined finished loweringPickering (Heresztyn and wines 1987; glucose others Villettaz 1999c). 1987; Glucoseing reduction GOX in is grape presently juice limited us- of to clarification white followed by grape enzyme varieties, reactionto as must yeast a first inoculation occur period prior forcose fermentation fraction in to grape commence.fermentable juice As sugars, represents the the approximately theoretical 50% glu- maximum ofproduction reduction total is in 50%, alcohol compared to winesIn made practice, from untreated some juice. inefficiencyported in alcohol glucose reductions for oxidation wines arises produced and using GOX-treated re- may dictate that thefor finished microbial stabilization, product thereby will leadingteration require to of pasteurization potential volatile loss flavorleaf or and al- area aroma compounds. to Adjustingfor crop moderating vine ratio the concentration is ofharvested fermentable an wine carbohydrates grapes in interesting (Stoll and viticultural otherspromising 2009; and intervention Whiting emerging 2010). This approachcentrations to managing attempts grape to sugar addressdrate con- the accumulation and imbalance the between developmentgrape carbohy- of constituents. sensorially However, important significantto research determine is the still optimumtion required of leaf leaf to removal from crop thelong-term vines ratios, impact relative to upon timing, fruit vine and location physiology. Juice and dilutionperformed loca- can the using only be a low Brixis grape not adulterate, a as permitted additionjuice, a of process by-product water for of wine grapeused juice production. concentrate, Low to is Brix more maintain commonly grape thermal wine distillation concentration techniques for during alcoholods reverse removal. Recent that osmosis meth- target prefermentation or productionalcohol strategies in for wines reducing have,minimize therefore, focused loss or upon alteration technologiesoff-flavor of that development. The desirable use organoleptic of qualities enzymeing and technology the for concentration lower- of fermentablelimiting sugars in alcohol grape juice production thereby methods prior and will to form theproduction fermentation basis options of in is discussion this for one article. prefermentation such Glucose oxidase 0.1%. Signifi- ± Early fruit harvest Juice dilution Glucose oxidase enzyme Modified yeast strains Arrested fermentation Pervaporation Osmotic distillation Solvent extraction Ion exchange Spinning cone column ´ acs and others 2007; Aguera and Vol. 71, Nr. 1, 2012 r sugars production extraction Reduced alcohol Nonmembrane Journal of Food Science Limiting alcohol production during fermentation by lowering fermentation Concurrent with Postfermentation Membrane separation Reverse osmosis Stage of wine productionPrefermentation Reduced fermentable Principle Technology R26 Pickering 2000; Smith 2002;others Tak 2010). Table 1–Technologies for reducingwine ethanol concentration and in fermented beverages (Duerr and Cuenat 1988; Production wine constituents (Aguera and others 2010).allow This winemakers information to will assess thethe relative technical technologies merits described of andmentation each make of of novel decisions winemaking regardingconcentration techniques imple- in for wine. reducing ethanol Prefermentation Technologies for Limiting Alcohol form the basis of discussionclassified in according this to article. the Technologies have stage been cally of used; wine that production is, they pre,However, are some concurrent, typi- well-established or “post postalcoholic fermentation. fermentation”such techniques, as low-temperature distillation, havecoholic been fermentation applied for during the al- ethanol removal of without approximately significantly 2% changing v/v the of concentration of other cant differences in flavorpresent intensities in and balances the areThis favored purportedly anecdotal sweet evidence spot shouldinvestigations alcohol be that levels considered demonstrate (Wollan in ethanol 2006). approximately light 1.0% difference of v/v thresholds recent (Yu are tion and methods Pickering 2008). thatwhile The are not produc- of exhaustive, most are relevance summarized to in the Table 1 wine and industry, generally wine sales compareders to may also full wish alcoholicfull-strength to wines marginally strength to reduce correct wines. alcohol balancestyle and concentrations Produc- between maintain in consistency vintages of oralcohol blends. reduction It can has2006). lead These also to sweet been an spots reportedlevel have alcohol that in been “sweet described a spot” when given (Wollan the wine alcohol is varied by as little as Technologies for reduced alcoholic wines . . . with certain positive health benefits (NationalResearch Health and Council Medical ethanol [Australia] concentrations 2009). arecountries Beverages also more thereby with favorably producing reduced excised some in most competitive advantages for be produced. Earlyorganoleptically grape undeveloped due harvest to reduced mayvelopment flavor in precursor result de- the in grapeslack wines prior of to that harvest, yeast-contributed are highrested flavor acidity compounds. fermentation levels, Comparatively, and leaving ar- high residual sugar levels in wines the concentration ofgrape fermentable harvest, juice sugars dilution, or innificant arresting levels juice of fermentation, unfermented while through sugars sig- of early remain in the the options wine that are some enable wines with reduced alcohol levels to R: Concise Reviews in Food Science lcncai.Ctls nyedgae h yrgnprxd omddrn lcs xdto HrmiradWlcx1981). Willcox and (Hartmeier oxidation glucose during formed peroxide hydrogen the degrades enzyme Catalase acid. gluconic of oxidation 1–Biochemical Figure of 1987). determination Villettaz techniques by 1987; laboratory (Heresztyn wine performed standard using monitor- be acidity Indirect titratable can and juice. pH oxidation grape is glucose of and of acidity preparations high ing GOX the by most affected in not of others present removal and is efficient peroxide, (Pickering for hydrogen required wine activity, finished enzyme Catalase adequate the 1998). an in provide suffi- alcohol to a of concentration ensure reduction glucose to with in required juice reduction be acid cient the therefore, to may, of due carbonate Deacidification 75% activity. to calcium rate up enzyme the by range, of reduced (O’Neil pH is inhibition activity production juice GOX acid grape gluconic optimum lower of corresponding considerably for 1998), the range others At pH 2006). and reported (Pickering the 6.0 to to 5.5 of range . . . wines alcoholic reduced for Technologies hscnrsswt h eotdtmeaueotmmfrGOX for optimum temperature reported the with contrasts This tatmeaueo 20 of occurring temperature oxidation a glucose at rapid more with demonstrated experiments juice Early grape ambiguous. are activity GOX for perature periods. prolonged for rates excessive at grape sparging of from oxygen arise Loss or potentially air 1999b). also may others components and and precursors volatile (Pickering storage wines medium-term to control short- than and during browning reactions to han- color susceptibility less reductively “pinking” have with do wines produced GOX juice. wines developed dled more than a color have yellow to reported golden con- is and wine resulting fermentation the during sequently precipitate 1987). (Villettaz do color phenolics brown a Oxidized of development and juice of constituents grape polyphenolic the of oxidation in result of also oxidation will the glucose, during during juice formation peroxide high the and of with oxidation, sparging glucose reported The been 1987). exces- (Heresztyn also levels with have aeration Problems evaporation and 1998). acid foaming others gluconic are sive of and rates rate mixing (Pickering the and production influence design, that sparger considerations rates size, Different important bubble juice. sparging, grape oxygen air of dissolved of treatment optimum GOX the for describes concentration that infor- available Little is ratios, concentrations. mation volume oxygen to dissolved area increasing surface thereby bubble maximizing limiting by and probably size activity, of bubble GOX the dispersion enhance into assist and will sparged bubbles, Agitation be oxygen treatment. must enzyme and during activity juice GOX for requirement tial feto xgno GOX. on oxygen of Effect feto eprtr nGOX. on temperature of Effect ◦ oprdt 30 to compared C β Dguoet -lcn-,-atn guoi acid (gluconic D-glucono-1,5-lactone to -D-glucose oeua xgni nessen- an is oxygen Molecular eot notmltem- optimal on Reports ◦ Hrstn1987). (Heresztyn C ciiy neial irba rwhmyas edcesdat decreased be also GOX may 20 for growth reactant microbial limiting oxy- rate Undesirable and activity. important temperatures lower an at is achieved concentration oxygen be gen dissolved can juice Higher the temperatures. in levels processing lower for ent rdcinrtsi O-rae uc ttmeaue f2 and 20 of temperatures at 30 acid juice GOX-treated gluconic in in rates difference production significant any demonstrated not have ciiybten3 n 35 and 30 between activity xds-rae rp uc Hrstn18;Vletz1987; Villettaz glucose 1987; from 1999c). (Heresztyn others made and juice Pickering wine grape corresponding redu- oxidase-treated in alcohol and percentages juice grape ction in rates oxidation 2–Glucose Table fgot tti temperature. this at growth of n tes19b.T oeaeti rbe,adt optimize to and problem, this moderate To 1999b). (Pickering others balance by of and prepared out wines wines the in render acidity may total juice GOX-treated The to composition. acid wine gluconic finished of acid for fermentable contribution problem gluconic of significant of fraction a levels is glucose High sugars the deacidified 4. of Table from conversion in from prepared given produced also wines com- is control The juices 3. and grape GOX and 2 Table wines of in and given position is juice wine treatment in GOX concentrations, from reduction arising alcohol acid resulting gluconic and reduction, composition, glucose of mary ait m/)()(/)(%) (g/L) reduction (%) acid (mg/L) reduction concentration variety Grape hsea 0 SN 23 NS NS NS 500 M Chasselas isig20 SN 36 23 NS 52 to 10 NS 36 2000 500 Riesling Riesling Muscat Muscat Muscat uller- Blanco Gordo Blanco Gordo Blanco Gordo Thurgau ¨ O rdcdwnsadtercomposition. their and wines produced GOX = ◦ ◦ Pceigadohr 98.Svrlavnae r appar- are advantages Several 1998). others and (Pickering C ,atog aywn piaeognssaeqiecapable quite are organisms spoilage wine many although C, o stated. not δ xds/aaaeGuoeGuoi Alcohol Gluconic Glucose oxidase/catalase lcoe yguoeoiaeadsbeun yrto to hydration subsequent and oxidase glucose by -lactone) o.7,N.1 2012 1, Nr. 71, Vol. Glucose 005 333 53 40 56 73 1000 87 2000 0 12 13 25 31 200 01 13.6 11 13 50 ◦ ONi 06,wieohrreports other while 2006), (O’Neil C r ora fFo Science Food of Journal sum- A R27

R: Concise Reviews in Food Science § 5 0 0 3 2 08 017 65 9 1 13 7 95 2 ...... Control wine with lower ethanol has been described; (Erten and Campbell 1 19 010 010 523 . . . . . § (% v/v) reduction (%) 210 0<1 0<1 7<0 882 17030 0 0 49 1 82 88 05610 05 3 2 510 ...... † were demonstrated to have an 43 39 S. cerevisiae 241284 170 209 S. cerevisiae 811 910 010 010 38 . . . . . Williopsis Saccharomyces cerevisiae strains for wine production. and 7<1 8<1 366 1 657 14033 0 0 93 2 31 127 25822 08 3 3 ...... 75 3 60 4 50 6 40 7 09 13 . . . . . pH acidity (g/L) 18 Pichia † Saccharomyces 03 83 03 23 13 . . . . . 784 289 7<0 896 33075 0 0 89 1 74 77 93710 07 3 3 ...... 2001). A problem of fermentation usingis novel or potential wild yeast off-flavor species developmentcharacters and (Heard undesirable 1999), organoleptic modified hence the development of genetically predictability than reliance upon2000). natural A difficulty fermentation faced (Pretorius bythe some production winemakers of and wine viticulturists grapescomponents is that and have accumulated a balance sugar.producers between to The flavor harvest requirement grapes forachieve at typical some high varietal levels of charactersalcohol fermentable produces concentrations sugar wines and to with undesirableLopes excessive palate and hotness others (de 2000).production Barros One of alcohol strategy in to these wines overcomewith is the lowered ethanol the production excessive selection during of fermentation. yeastability Some strains vari- in ethanol productionwine by starter different cultures of commerciallyhowever, the available difference in final ethanol concentrationsin determined one controlled experiment wasThe less selection than 1% of v/v yeasts (Jensonproduction other 1997). rates than for grapecohol juice wines fermentation produced by is the possible. fermentationusing Low of strains al- oxidized of grape juice acceptable, albeit different palate structureities, and than organoleptic wines qual- produced with Wine analysis Wine analysis after alcoholic after malolactic 57 6 37 6 30 8 25 9 00 16 . . . . . pH acidity (g/L) † 03 03 05 3 25 3 73 . . . . . 710 887 372 91 6715028 72 0 0 77 3 226 89722 49 2 3 ...... Deacidified GOX-treated Control Decreased 63 6 40 7 31 8 20 10 84 19 . . . . . ◦ 03 03 25 3 02 a.u. 1 g/L 2 g/L 3 Vol. 71, Nr. 1, 2012 . . . . mg/L 95 r ¨ uller-Thurgau and Riesling wines prepared from GOX-treated deacidified juice (Pickering and others ¶ ‡ † † ‡ 2 Composition of M Glucose oxidase/catalase treated Chasselas juice and reduced alcohol wine production (Villettaz 1987). 2 2 – – Journal of Food Science Selection of specific yeast strains and their use as starter cul- ¨ uller-Thuragau To t a l S O Bound SO Gluconic acidTartaric acid g/L g/L <0 1 EthanolGlucoseFructose % v/v g/L g/L 84 89 6 Titratable acidity A420A520Free SO 0 0 Total hydroxycinnamates Titratable acidity Total flavonoids pHpH 4 5 EthanolBrix % v/v 6 Analysis at time of bottling. Caffeic acid equivalents. As tartaric acid. Corrected for gluconic acid/lactone. As tartaric acid. § Component juice juice juice alcohol wine ‡ ¶ R28 † 1999a, 1999b). M 003 † Table 4 Table 3 Glucose oxidase/ catalaseconcentration (mg/L) Contact time hours pH acidity (g/L) Titratable Titratable Titratable Alcohol Alcohol Production Use of novel yeast strains ous outcome of GOXthese treatment is wines an to increased undergocreased susceptibility premature flavonoid production for browning (Pickering consistent and others with1999c). 1999a, in- 1999b, Fermentation Technologies for Limiting Alcohol associated with the production ofjuice wines is from GOX-treated the grape formationpounds, of resulting in substantial significantly quantitiesthan higher of in sulphur control carbonyl wines dioxide com- (Pickeringthe binding and others total 2001). concentration Consequently, microbial of stabilization sulphur in dioxide GOXceed wines necessary legal may limits to approach (Pickering achieve or and even others ex- 1999a). A further deleteri- Technologies for reduced alcoholic wines . . . GOX activity, deacidificationcarbonate of prior the to grapescheme GOX for juice treatment the production may of with reduced bement alcohol calcium wine of necessary. by white GOX A treat- grape typical juice is shown in Figure 2. A particular issue Treatment Juice analysis fermentation fermentation tures for themon consistent winemaking manufacture practice. ofwine The production a also use ensures improved wine fermentation of reliability and style specific is yeast a strains com- for Riesling 50 2 100 2 500 2 500 15 R: Concise Reviews in Food Science et eesgicnl ifrn oatpclgaejieo must or juice grape typical experi- a these to in different conditions significantly culture were However, ments glyc- formed. of be levels higher to enabled cultures erol 2001) aerated 1996, in phos- others concentrations and triose (Compagno glucose or low at 1990) isomerase others phate and (Drewke dehydrogenase cohol targeted. biochemical been that have associated products balance and redox gene in pathway involved of pathways glycolytic wines number the of A with concentration 1997). interact identified ethanol others final been the and has decrease (Bartowsky biomass to strategy increasing a production or as to metabolites, ethanol from other compounds of carbon grape divert that eeial engineered Genetically . . . wines alcoholic reduced for Technologies al okwt igegn uain oionye fal- of isoenzymes to mutations gene single with work Early of strains yeast specific engineer to potential The .cerevisiae S. .cerevisiae S. ltn tao rdcin hl anann eo aac and balance redox maintaining while mod- production, in promise ethanol some ulating show growth yeast en- during oxidase expressed Trans- zymes (NADH) 2009). hydride dinucleotide others adenine and nicotinamide oxygen-dependant (cytosolic) Hou soluble 2006; water 2006b; of others incorporation 2006a, genic and others (Geertman pathways and of of flux Heux suite the a altering thereby through and carbon cell yeast the within balance redox strains. yeast wine specific in encoding of interest genes development of the the enzymes of redi- and manipulations genetic synthesis, to for ethanol methods potential from flux with carbon pathways rect in with metabolic important interpreted specific were experiments be identifying early should these Nonetheless, results caution. therefore and fermentation, oercnl,cfco niern a enue omodify to used been has engineering cofactor recently, More o.7,N.1 2012 1, Nr. 71, Vol. eue loo ie sn lcs oxidase. glucose using wines alcohol reduced of production for scheme 2–Processing Figure r ora fFo Science Food of Journal R29

R: Concise Reviews in Food Science ˜ no )-2, 3- ´ alez and R has been ,3 ´ ereau-Gayon R S. cerevisiae )-acetoin to (2 S were developed that included overex- strains with a downregulated PDC gene acetoin, and (3 3R)- S. cerevisiae Saccharomyces cerevisiae A novel method to overcome the deleterious effects of high While genetic engineering has shown significant promise as a an acetaldehyde dehydrogenase-6 (ALD-6)stantially gene lower deletion, ethanol a and sub- demonstrated acceptable (Eglinton acetate and concentration others was tration 2002). of However, other the sensorially concen- important metabolites,hyde such as and acetalde- acetoin, wereconsidered significantly to be in acceptable excess fornipulations of wine of products. concentrations Further genepressed ma- 2, 3-butanediol dehydrogenaseers (BDH) 2009), (Ehsani and anconversion oth- NADH-dependant of ( reductase responsible for the and others 1999). and overexpressed GPDH-1 were developedproduction to at enhance the glycerol expensePDC of activity was ethanol reduced (NevoigtGPD-1 to and less overexpression Stahl than resulted 1996). one inhowever, the fifth end a of products, wild acetate 45% and typealternate reduction acetaldehyde and derived redox in from reactions, ethanol; and were Stahl subsequently 1996). increased (Nevoigt acetate levels inpressing fermentation yeasts has conducted been by developed. GPDH-2 By overex- using a yeast strain with Controlling the fluxucts of not metabolic specifically intermediatesstrains targeted and remains an end by important prod- geneticable, barrier must manipulations that, be in controlled while in notthat yeast order insurmount- are to achieve organoleptically commercialof acceptable. products one Deletion gene or product overexpression can significantly alter a suite of metabolic technology to enable selection offor yeast expression strains of specifically desirable tailored take traits, of a this number technology of for barriers regular wine to production the are up- apparent. Further limitations of gene technology formanipulations yeast strain butanediol and meso-2-3,butanediol, respectivelyothers (Gonz 2000). A summaryin of cellular metabolite redox concentrations balancetargeting involved GPDH, using ALD, and genetically BDH modified expression5 is and yeast presented important strains in biochemicals involved Table in theirin formation Figure is 3. shown Clearly, anymanipulations causes attempt numerous to biochemical pathway redirect interactions. Consequently, carbon redox imbalance flux within by the gene celland must arising be from corrected, alternate biochemicalproduction of pathway a modulation suite is of the sensoriallyexceed important desirable compounds organoleptic that may concentrations in wine. (Michnick and others 1997;Lopes Remize and others and 2000; others Cambon andtion 1999; others by de 2006). yeasts Acetate Barros during forma- fermentationof may acetyl-CoA occur or either the pyruvate by dehydrogenaseway hydrolysis (PDH) in bypass which path- pyruvate is decarboxylatedby to oxidation to acetaldehyde acetate followed (Remize and othersand 2000; Rib others 2006). Thepyruvate enzymes decarboxylase involved (PDC) in thenase and PDH that an belongs bypass acetaldehyde to are enzymes. the dehydroge- aldehyde The dehydrogenase (ALD) ALD groupextensively group of characterized of with 5 enzymesnated, isoforms in and (ALD2-6) being the desig- mostALD-5 important (mitochondrial) of and these ALD-6stitutive during enzymes. (cytosolic) The fermentation as expression are both ofrepressed and are isoforms therefore these con- ALD2-4 enzymes do is notlar glucose play redox any balance role in during cellu- grape juice fermentation (Navarro-Avi during with the major- S. cerevisiae S. cerevisiae Vol. 71, Nr. 1, 2012 r ´ ereau-Gayon and others 2006). The , any influence upon the flux of these com- + ´ evant 1991). Journal of Food Science As glycerol and ethanol production by yeasts during fermenta- Glycerol formation arises from reduction of the glycolytic in- Most effort in recombinant technologies for manipulation of Genetic manipulation of yeast strains to overexpress either R30 regeneration of NAD others 1997). Osmoprotectiontion in by upregulated the GPDH-1 early expressiona has stages more been of shown significant toof fermenta- have role redox in imbalance by glycerol(Remize GPDH-2 and production others during 2003). than anaerobic correction fermentation tion are important regulators of cellular redox balance through the and subsequent dephosphorylation.alyzed These by an 2 NADH-dependentgenase reactions glycerol-3-phosphate (GPDH) are dehydro- cat- andrespectively. Two a isoforms of specificdesignated GPDH GPDH-1 glycerol-3-phosphatase and have (GPP), GPDH-2 beenencoding with these described expression isoenzymes regulated of and by theosmoprotection the genes yeast and requirement for redox balance, respectively (Michnick and formation involve oxidation of NADH,arises the redox from imbalance anaerobic that glycolysisration is and corrected glucose (Scanesof repression and redox of balance others respi- that 1998). isfunction It considered of is the glycerol the formation most important correction (Michnick and biological others 1997). termediate dihydroxyacetone-phosphate to glycerol-3-phosphate pH, sugar concentration, nitrogen source, and sulphurels dioxide (Scanes lev- and others 1998;benefits Remize of and others glycerol 1999). formation Theare for major 2-fold. yeast cells Glycerol during isbiomass fermentation normally formation produced at by commencementcells of from fermentation the high to osmolar protect venting concentrations cellular of sugars, dehydration. thereby pre- Also, as the reactions for glycerol 4% and 10% of grapeproduction juice during carbon is fermentation normally directedity by to being glycerol produced during(Scanes the and initial others stages 1998; of Rib final biomass formation concentration ofranges glycerol between in 4 finishedstrain and and dry a 9 range wines g/L of environmental normally and signals including is temperature, dependent upon the yeast investigations involve the utilization of relatively lowconcentrations carbohydrate relative to grape juice or must. wine yeasts has targeted increasingpense glycerol of production at ethanol the(Nevoigt and ex- and has Stahl required 1996;Lopes multiple Michnick and and gene others others modifications 2000; 1997; de Geertman Barros and others 2006). Between Technologies for reduced alcoholic wines . . . avoiding unwanted accumulation of sensoriallyas active compounds carbon flux isicant redirected disadvantage through of multiple cofactor pathways.soluble engineering A oxygen is signif- to the requirement be for supplied during fermentation and current compounds in winemay at be detrimental levels to exceeding perceived wine1988; their quality Eti (Boidron sensory and others threshold pounds will alter the concentration ofthat a are range also of other involved metabolites succinate, with acetoin, redox balance. diacetly, Acetaldehyde, andmost acetate, important 2,3-butandiol of appear theseRemize to metabolites and (Michnick be others and the 2000; othersand Taherzadeh 1997; others and 2006; others Ehsani 2002; and Cambon others 2009). The presence of these erol formation andin decreased acetate, ethanol succinate, production was acetoin, increased acetaldehyde, and 2, 3-butandiol GPDH-1 or GPDH-2 hastween resulted 19% in diminished and ethanolin 22% of grape be- juice in fermentations. model Concomitant solutions with increased but glyc- significantly less R: Concise Reviews in Food Science ehooisfrrdcdachlcwns... . . wines alcoholic reduced for Technologies

Table 5–Changes in metabolite concentrations of yeast cultures with modified gene expression for glycerol production. Concentration of metabolites Yeast strain Modified gene Ethanol reduction Ethanol Glycerol Acetate Succinate Acetoin 2, 3-butanediol Acetaldehyde designation product expression Growth media % g/L g/L g/L g/L g/L g/L mg/L Nevoigt and Stahl (1996)† Glucose 18 g/L Wild type Control 7.9 0.6 0.20 NS NS NS <5 GPD-1 GPDH-1 overexpressed 35 5.1 4.1 0.58 NS NS NS 361 pdc PDC downregulated 29 5.6 2.9 0.29 NS NS NS <5 pdc GPD-1 PDC down regulated 45 4.3 5.1 0.40 NS NS NS <5 GPDH-1 overexpressed

Michnick and others (1997) Glucose 100 g/L, pH 3.3 V5/pVTU Control strain 46.8 4.3 NS NS NS NS NS GPD1 V5/GPD1 GPDH-1 over expressed 22 36.6 14.0 NS NS NS NS NS V5/pVTU Control strain Glucose 200 g/L, pH 3.3 89.2 7.1 0.52 0.25 <0.1 0.90 <100 GPD1 V5/GPD1 GPDH-1 overexpressed 19 72.5 28.6 1.60 0.54 6.10 1.30 220

Remize and others (1999) Glucose 200 g/L pvt100-U-ZEO R Control strain 88.4 7.4 0.42 0.40 0.00 0.24 0.01 pvt100-U-ZEO-GPD1 R GPDH-1 overexpressed 3 85.7 16.5 1.18 1.11 0.06 1.92 0.04

de Barros Lopes and others (2000) Chardonnay juice 21.8 ◦Brix, pH 3.16 AWRI 838 Control strain 129.8 7.9 0.58 0.39 NS NS NS AWRI 838 GPD2-OP GPDH-2 overexpressed 4 124.0 16.5 1.02 0.65 NS NS NS Eglinton and others (2002) Glucose 80 g/L GPD2 ALD6 Control strain 34.1 5.1 0.66 0.59 NS NS 0.64 GPD2-OP ALD6 GPDH-2 overexpressed 24 26.0 13.4 1.42 0.83 NS NS 8.36 ALD6 normal expression o.7,N.1 2012 1, Nr. 71, Vol. GPD2 ald6 GPDH-2 normal 9 31.1 6.0 0.20 0.59 NS NS 0.79 expression ALD6 deletion GPD2-OP ald6 GPDH-2 overexpressed 20 27.3 16.3 0.36 0.88 NS NS 8.79 ALD6 deletion Cambon and others (2006) Glucose 200 g/L; malic and citric acid 6 g/L; YAN 460 mg/L VL1 Control strain 97.2 6.5 0.57 0.31 NS 1.14 16 r ora fFo Science Food of Journal VL1ald6 ALD6deletion <1 96.6 7.4 0.19 0.29 NS 1.30 8 VL1 GPD1 GPDH-1 overexpressed 17 80.2 24.4 2.64 0.48 5.4 4.01 105 VL1 GPD1 ald6 GPDH-1 overexpressed 23 74.7 26.8 0.62 0.62 6.2 6.05 182 ALD6 deletion K1M Control strain 93.3 5.8 0.38 0.31 NS 0.61 39 K1M ald6 ALD6 deletion 93.6 6.8 0.10 0.43 NS 0.52 18 K1M GPD1 GPDH-1 overexpressed 11 82.6 18.2 0.98 0.78 4.2 4.85 183 K1M GPD1 ald6 GPDH-1 overexpressed 16 78.8 21.9 0.43 0.88 5.8 3.93 228 ALD6 deletion (Continued) R31

R: Concise Reviews in Food Science The separation of 2 solutions of unequal The removal of ethanol from wine following the completion Reverse osmosis. Some of the gene inserts used to induce overexpressed gene From the explanation outlined above, it should be easy to see

of fermentation canof be thermal achieved distillation either processes,transport through with of or ethanol the across without a application vacuum, semi-permeableVarious barrier or technologies or the in membrane. whichtive a removal membrane of is ethanol used fromrely for beverages upon the have been selec- molecular developed permeationwith that of high ethanol concentration, to fromtration. a the The stripping feed most stock phase prevalentremoval membrane-based with of technology low organic for concen- constituents the sis, from and beverages emerging ispervaporation, reverse technologies, are osmo- such still nonmainstream. as osmotic distillation and possible future use of gene technologythe in identification the and wine comparison industry of will fermentationyeast be strains performance traits of bred by selective breeding techniques. Postfermentation Technologies for Removing Alcohol Membrane transport processes intermediates and end productsnipulated during yeast fermentation cells attempt asSome the to metabolic ma- compensate traitsinteraction altered are metabolism. of encoded several bywithin gene a the systems yeast suiteoenological genome of on outcomes (Pretorius genes by different genetic 2000). orbe chromosomes engineering Achieving possible the with may desirable approaches thereforeA targeting not a complicating limited factor numberthe in of failure genes. genetic of manipulations some transgenic ofa strains wine timely to yeast manner complete fermentation is (Remize2003; in Heux and and others others 2006b). 2000; Remize and others products may not be stabledisadvantage within is the the altered loss strain.ing of A fermentation the significant plasmid (Remize insert andothers others from 2000). yeast 1999; cells de Thus, dur- Barros incorporationtaining Lopes of overexpressed and genes, stable or plasmidduction gene inserts of deletions con- specific limitingachieved the enzymes in order pro- that to produce alter consistentwith and metabolic these reliable fermentations flux, yeasts.consumers Most must importantly, to be however, foods ismodified that the organisms (GMOs). contain attitude The of or useis of a produced GMOs major in cause using food of concern genetically products led and to mistrust the among introduction consumers of andbetween some has trading economic restrictions zones, ofof these labeling goods GMOs requirements in stating foodby production, the retailers and, use to in stock someGMOs products instances, or that the their cannot refusal metabolic be products proven to (Burton be and free others of 2001). A known as reverse osmosis.dilute Effectively, solution the reduces concentrationincreases. while of that the of the concentrated solution how this phenomenoncess. could In be fact, exploited simply as adjusting a the filtration pore pro- size of the membrane solute concentration by aa semi-permeable concentration membrane or establishes osmotic pressure pressure. gradient Incess such between of a them osmosis system, from knownmembrane the water in solution as will order of to low movepressure concentration restore by (greater equilibrium. across a than the However, if the pro- osmotic sufficient concentration pressure) side, is solvent can applied move on out ofmembrane the this high to solution across the the low concentration solution, in a phenomenon

eyrgns;NS dehydrogenase; o stated. not =

yuaedcroyaegn;GPDH-1 gene; decarboxylase pyruvate PDC lcrl3popaedhdoeae2gn;ALD6 gene; 2 dehydrogenase glycerol-3-phosphate GPDH-2 gene; 1 dehydrogenase glycerol-3-phosphate -2,3-butanediol )- R ,3 R (2 BDH1 gene; 6 dehydrogenase acetaldehyde = = = = =

ovre rmmmol/L. from Converted

†

deletion

vrxrse ALD6 overexpressed

223 222 221 89 206 . . . 120 7.4 0.6 0.9 0.65 32 97 18 BDH-1 and GPDH-1 5ad P1BDH1 GPD1 ald6 V5 , ,

deletion

vrxrse ALD6 overexpressed

5ad P1BH PH1adBDH-1 and GPDH-1 BDH1 GPD1 ald6 V5 79 007 . . . 120 8.0 0.5 0.9 0.70 30 98 17

L6deletion ALD6

5ad P1GD- overexpressed GPDH-1 GPD1 ald6 V5 69 206 . . . 140 2.5 5.9 0.9 0.65 32 99 16

5ad L6deletion ALD6 ald6 V5 1 .405001310 1.3 0.0 0.5 0.14 9 117 1

<

5Cnrlsri 1 .304000910 0.9 0.0 0.4 0.63 7 118 strain Control V5 Vol. 71, Nr. 1, 2012

r haiadohr 20)Guoe20g/L 240 Glucose (2009) others and Ehsani

L6deletion ALD6

CGD l6GD- overexpressed GPDH-1 ald6 GPD1 BC 87. 6905 .49550 320 5.09 9.5 0.74 0.50 26.9 75.5 18

CGD PH1oeepesd1 271. .605 . . 95 3.9 2.9 0.58 1.36 16.5 82.7 10 overexpressed GPDH-1 GPD1 BC

Cad L6dlto 047100 .4N .910 0.99 NS 0.64 0.05 7.1 90.4 2 deletion ALD6 ald6 BC

CCnrlsri 247203 .7N .59 1.45 NS 0.37 0.38 7.2 92.4 strain Control BC

einto rdc xrsinGot ei / / / / / / mg/L g/L g/L g/L g/L g/L g/L % media Growth expression product designation

es tanMdfidgn tao euto tao lcrlAeaeSciaeAeon2 -uaeilAcetaldehyde 3-butanediol 2, Acetoin Succinate Acetate Glycerol Ethanol reduction Ethanol gene Modified strain Yeast ocnrto fmetabolites of Concentration

Journal of Food Science al 5–Continued Table R32 Technologies for reduced alcoholic wines . . . R: Concise Reviews in Food Science iue4Sprto aaiiiso ifrn ebaessessoigtpcloeaigpesr i aetee) RO parentheses). (in pressure operating typical showing systems membrane UF different nanofiltration; of capabilities 4–Separation Figure in production ethanol and flux carbon modulating for targeted enzymes specific and pathways 3–Biochemical Figure . . . wines alcoholic reduced for Technologies tes20,Esn n tes2009. others and Ehsani 2007, others BDH ALS decarboxylase; pyruvate GPDH isomerise; phosphate triose = ,-uaeildhdoeae DR dehydrogenase; 2,3-butanediol = lrfitain MF ultrafiltration; = ctlcaesnhs;ALD synthase; acetolactate = lcrl3popaedhdoeae GPP dehydrogenase; glycerol-3-phosphate = microfiltration. = ictlrdcae oie rmRmz n tes20,Rib 2000, others and Remize from Modified reductase. diacetyl = ctleyedhdoeae PDH dehydrogenase; acetaldehyde = lcrl3popaae ADH glycerol-3-phosphatase; o.7,N.1 2012 1, Nr. 71, Vol. = yuaedhdoeae DS dehydrogenase; pyruvate ra-ao n tes20,Crirand Cordier 2006, others and ereau-Gayon ´ r = ora fFo Science Food of Journal acaoye cereivisiae Saccharomyces loo eyrgns;PDC dehydrogenase; alcohol = ees soi;NF osmosis; reverse = ictlsynthase; diacetyl .TPI R33 = = =

R: Concise Reviews in Food Science In a reverse osmosis process for alco- C (Smith 1996). Operating conditions ◦ 200 Da (Catarino and others 2007) so that water and ethanol, velocity under pressure. Some liquid passes throughbut the the membrane solids or materialsnominal with molecular molecular weight weight cutoff higherwill (NMWCO) than be the of swept the alongRecycling membrane in will the ensure stream thatmembrane of more during permeate feed each will acrossthe cycle pass the feed until through membrane. is achieved. the the Inconfigurations order desired to have concentration effectively been do of developed. this, These several(also module include known the as flat platewound sheet and configurations. frame), The spiral tubular, wound hollow configurationmost fiber, makes economic and the spiral useflat of membranes space rolled for upThe a together original given like membrane space acollection area, between cigar channels being (Pretorius the and 2000). membranesthe the membranes serve new becomes space the as feed generated permeate channel. fromApplications winding and limitations. hol reduction in wines,alcohol the content. feed is This the wine(40 regular is atm) wine through pumped with a at normal membranesult pressures module in up and elevated to such temperatures pressures 4excessive temperature can at MPa arising re- the from high membrane pressures,are heat surface. exchangers typically To avoid a componentperatures around of 20 the to 22 apparatus with operating tem- Figure 5–Schematic of a typical asymmetric composite membrane(1) showing: thin skin of porous membrane;(3) (2) polyester polymeric support. micro-porous support; and being small molecules, passmeate through the stream. membrane The into retentate the per- is redirected to the feed tank and must be balanced betweenat gaining higher efficiencies pressures inlower and temperatures permeate (Catarino aroma flux and retention, othersers 2007; 2009). which A Labanda membrane is and is oth- selected< improved with at a low NMWCO, typically Reverse osmosis membranes can Vol. 71, Nr. 1, 2012 r ) is called the flux and is dependent on the 2 − m 1 − Journal of Food Science During membrane processing, the feed is separated into Reverse osmosis, such as other membrane techniques, operates As a membrane technology, reverse osmosis requires low-energy The first patent for the application of reverse osmosis in al- R34 square (L h Thus, not only waterular or weight substances may with pass comparably throughbut low the other molec- membrane as solutes in andweights. reverse solvents osmosis in gradation of size and molecular 2 streams: retentate (concentrate) and permeateumetric (filtrate). throughput The of vol- the membrane surface per hour, per meter Technologies for reduced alcoholic wines . . . material and the applied pressurebrane give filtration rise processes to with a increasingreducing spectrum solute operating of permeability mem- pressure) (and suchtion, as ultrafiltration, reverse and osmosis, nanofiltra- microfiltration, respectively (Figure 4). under the principle ofuid cross flows or parallel tangential or flow, tangential whereby to the the liq- membrane surface at high polymer (membrane material) withlectivity. Such the membranes required give permeationvery good se- durable flux under the characteristics high-pressuresis. application and These of are membranes reverse osmo- are also cleanablerestore and initial allow flux back flushing rates to on by the destabilizing membrane any surface. build up of materials aration of uranium isotopesmembranes (Gibson are 1986). the The asymmetricthin most (heterogenous) skins successful types, of whichers membrane are of material polymeric bondedbrane support configuration to material (Figure one 5). toare Thin-film or create composite very more membranes a commonly lay- compositeand used porous mem- where structure is a chemically polymer bonded with to a high very strength thin film of Membrane types and configurations. be made of severalregenerated different cellulose, materials synthetic polymers, including and celluloseand ceramics Hartel acetate, (Heldman 1997; Westbrook 1989). The cellulosic materialsas are durable not and give lowmers, flux rates which compared are to also theand more synthetic durable, selective. poly- are Ceramics, also while expensive, very being designed strong originally for sep- technology or arrested fermentation,produced by the reverse osmosis reduced usually alcoholcomparable have flavor wines to and the aroma regular profile (Bui wines and from which others they 1986)process, were as and obtained no water phase andremoved change from alcohol the is are feedstock. associated largely with the the only components juices (Baldwin 1998; Smith 2002). input, operates at ambient temperatures,trol allows reproducible over con- separations,other requires additions, no and is disposable(Gibson easily filtration automated 1986). media for Specifically, continuous or inproducing operation comparison low to alcohol other wines methods such of as distillation, spinning cone coholic beverages wascompany obtained Lowenbrau in by 1975 for the thewine dealcoholization (Meier West of 1992). German beer Other and applicationsproduction brewing of include reverse osmosis removal in of wine tration colors (as and an flavors, alternativeproducts must to such concen- chaptalization), as development aperitifs,cipitation of wine and new stabilization deacidification (removal against of tartrate volatile pre- acids) of grape applied pressure and totalresistance is membrane a resistance. composite Theviscosity of of membrane permeate, factors pore that size, extentbetween limit of membrane permeation fouling, material and such interactions and as area, feed. the The greater larger the the filtration membrane capacity. R: Concise Reviews in Food Science hs fdgse uewtr evprto ssa nr a ihwtrvpr tao irtstruhtemmrn nagsospaeand phase gaseous a in membrane the through migrates Ethanol 1998. vapor. others and water Hogan with from gas stripp Adapted a permeate. inert employs as an distillation phase Osmotic uses stripping membrane. the pervaporation permeable within semi water: recondenses a pure across differential degassed pressure of vapor by phase removal ethanol of principle 7–Basic Figure a distil through thermally (1) to pressure by-product. used a is high as (5) collected under column (7) pumped rectification ethanol A and is wine (4). Wine the retentate to and process. back (3) added osmosis permeate (6) reverse streams, water 2 the closed-loop with into a permeate separated the is using and wine (2), of membrane semi-permeable dealcoholization for 6–Schematic Figure separate others to and permeate, the (Bui of unit unit rectification permeable commonly, this ethanol More from 1986). the supplying im- filtrate feed ethanol the the an redirecting to with one and by parallel, membrane possible in permeable units is osmosis or wine reverse has juice 2 dealcoholized using Brix also with is low This pressure wine, of tank. production osmotic alcohol-enriched simultaneous the feed The since the reduced. permeation also in increasing of content effect alcohol added, the juice con- the Brix volume lower low the more keep the the Basically, to process. be by-product the (wine) may during a feed juice stant juice, the Brix to Brix low added Alternatively, low continuously concentrate. original of juice the addition grape to the of restored is by Wine content 1992). water (Duerr Meier concentrated 1988; and Cuenat dealcoholized and continuously is wine the . . . wines alcoholic reduced for Technologies soi shge n trmvlo tao oblw04%v/v (Pilipovik 0.45% removed below ethanol to of liter ethanol per of electricity removal more at reverse consumes and of cost higher capital is the wines, osmosis alcohol low of preparation or tion allowing v/v, thereby % 0.5 wines, than alcohol less low to of flexibility. v/v production range 15% con- wide to alcohol 12% a reduce about to producing from used shown wine be is in can system tent osmosis closed-loop Reverse a 6. within Figure (Smith process in products typical wine a from now and removal are 1996) ethanol apparatus for Such used addition. the juice without commonly volume Brix feed wine low the maintains for system to requirement closed-loop back a component in water tank distillation the thermal redirecting using and possible is processes content water the and ethanol ncmaio oohrcnetoa ehd fdealcoholiza- of methods conventional other to comparison In o.7,N.1 2012 1, Nr. 71, Vol. r ora fFo Science Food of Journal R35 ing

R: Concise Reviews in Food Science The spinning cone column (SCC) is a device used to extract final concentration of 0.5% v/vtention in of chardonnay 80% resulted of in(Karlsson the and the re- Tragardh concentration 1996). Clearly, of furtherand research most the is aroma required retention compounds ofporation aroma becomes more compounds widely improved, adopted before forin moderation perva- beverages. of ethanol volatile flavor components fromconsists a of a liquid vertical or shaftporting rotating slurry. up at The approximately to 350 column 22 rpm,each inverted sup- pair (pointing of downwards) cones. cones,casing Between there of is the a column fixed (Figureof inverted the 8). cone, column The attached into liquid to the feed the 1st is spinning fed cone. to A the film of top liquid is flung Figure 8–Mechanical layout of the spinningin; cone 2: product column out; (SCC). 3: 1: gas product in;7: 4: rotating gas out; cones. 5: Courtesy rotating Flavourtech, shaft; 6: Lenehan stationary Rd., cones; Griffith, Australia. Spinning cone columns Vol. 71, Nr. 1, 2012 r Osmotic distillation, also referred to as ´ acs and others 2007). One report in which a hy- Journal of Food Science ´ acs and others 2007). The basic principles of ethanol removal Pervaporation technologies for ethanol removal have not been The nature of the stripping phase determines the process; os- Emerging membrane technologies: osmotic distillation permeate (Tak drophilic membrane was employed for ethanol reduction toR36 a as widely adopted asfew osmotic reports distillation in or thefor reverse literature osmosis, the describe and treatment thehigher of use temperatures wine. of required this Thismeation to technology may compared achieve possibly to effectivevation be ethanol osmotic will attributed per- also distillation to increase as the temperature flux ele- of aroma compounds to the 98% (Diban and othersto 2008). the Compound polarity losses and werecreased attributed with volatility residency of time for the treatment,ethanol thus compound potentially removal. and limiting Difference therefore testing in- not detect with significant untrained differences between the panels wines11.3% could at v/v 13.35 illustrating % and the potentialtion for for the the use removal of of osmotic small distilla- fractions of alcohol from beverages. improved from lower operating temperatures (Varavuth and2009). others As the vaporaroma pressure compounds differential is across the generally much membraneand lower for subsequent than ethanol, aroma the lossesminimized. flux of In a important pilot-scale flavor operation,moved compounds from 2 a are % merlot v/v wine and ofimportant the ethanol to loss was of wine re- compounds, flavour, considered to the permeate varied from 0.9% to considerably lower and are mostly retained withinbranes the wine. used Mem- for osmoticpolyethylene, polypropylene, polytetrafluoroethylene, or distillation polyvinyl are usuallydifluorides constructed in from varyingethanol pore flux sizes. is Compared considerablyquired to for lower the reverse equivalent and ethanol osmosis, removal; thus however, energyarise savings longer from times the considerably are lower re- pressures and product integrity Tak from wine byFigure establishing 7. Osmotic a distillation membranes vaporlar, have hydrophobic, properties, differential apo- which are are essential illustratedthe as this in flux critical of factor determines ethanolvor components from have lower retentate vapor to pressuresthan in permeate. ethanol ethanolic Aroma itself; solutions and thus, fla- the flux ratios of these compounds are a process of evaporationsion at of the the vapor wine acrossstripping the membrane membrane, phase. interface, and condensation diffu- into the motic distillation employs degassed pure1998; water Diban (Hogan and and others 2008), others whereas pervaporationan makes inert use of gas containing water vapor (Karlsson and Tragardh 1996; and pervaporation. evaporative pertraction(Diban and or others 2008) and isothermalin pervaporation that share similar selective membrane processes removal ofof ethanol, distillation a arises from vapor theally pressure establishment has differential hydrophobic across properties. Thus, a ethanol membrane, removal occurs which as usu- already being employed for improvingviability efficiency of and the maintaining winepossible industry. Apart uses from in dealcoholization, the other treatment of wine saline industry water for include irrigation,ter amelioration and to treatment of reduce of costs wine, waste wa- ofdifferent waste configuration disposal. Each of process equipment requiresprocessing slightly and of various permeate. changes to the Technologies for reduced alcoholic wines . . . and Riverol 2005). However, laborlow, and especially other since operating energy costsoperating are is costs would used also more dependosmosis, efficiently. as on part Savings the of plant on the capacity. family Reverse of membrane filtration techniques, is R: Concise Reviews in Food Science r eoee eaaet h rae rdc.TeSCcnbe can SCC The product. components treated volatile the and to (8) va- cyclone separate gas recovered condensate a stripping are to the fed column, are the the leaving pors through On back (3). passed exchanger once recovered heat remainder discharge the (5). product prod- with SCC (7), the the heater of of the reinjection portion into a a through of fed discharge treatment uct and from obtained (3) is exchanger gas warmed Stripping is heat product regenerative the (1), a efficiently tank function in feed to the leaving process After the 10). for (Figure required Other are constant 1999). items Sykes hence and ancillary (and (Harders pressure column constant the within almost transfer temperature) mass at the occur enables turn to in process pres- this that and minimized, ensure are cones drops fixed sure and rotating between clearances able iue9Cosscino oesoigfist raetruec.1 ttoaycn;2 oaigcn;3 n :rttn hf.Cuts Flavourtec Courtesy shaft. rotating 4: fin; 3: cone; rotating 2: cone; stationary 1: turbulence. create to Rd. fins Lenehan showing cone of 9–Cross-section Figure Typical 30 temperature. approximately are reduced temperatures a column at and off feed evaporated be will ponents within gas stripping and liquid both ensures column. for also the into cones length product of path volatile number adequate and of considerable an rotating The transfer the gas. mass of stripping a surfaces the upper enhance is the liquid that on the cones film that fixed thin fact a the and as turbulence out its this spread is spinning Along It the of 9). column. underside (Figure the the cones considerable to to attached of exposed fins by top is caused gas the turbulence stripping it at the components upward, collected path volatile tortuous is heater with up, a along picked through gas, has discharge The reinjection. product to the from and prior of generated rotating portion be can the a vapor between redirecting stripping voids alternative An the cones. through fixed passes and column extract. materials more coffee different to as wine) of such example, range materials (for a viscous products handle low-viscosity typically from can is feed, SCC time as resident The the s. thin, and quite 20 low is around is film volume liquid holdup the liquid As liquid the column. the until the of times bottom several the repeated spinning reaches 2nd is movement the onto the the passes and a under then cone, as cone liquid migrate the The gravity. of and of center cone influence the fixed toward a and cas- downwards onto the film drop of thin then surface will inside liquid the The onto ing. action centrifugal by outwards . . . wines alcoholic reduced for Technologies h ounoeae ne eaiepesr,s oaiecom- volatile so pressure, negative a under operates column The the of base the to admitted is nitrogen, as such gas, stripping A ◦ .Reason- C. rt a icltdbtenteSCadafretto vessel, fermentation a and SCC The the 1996). between Pyle circulated and was (Wright broth broth yeast fermenting a from moval 2009). others akgn nbe ag fpoutsye ob developed. be to styles product of and range filtration, a to enables prior packaging specifications product and final dealcoholized juice to or concentrates the juice wine, full-strength with with Blending aroma constructed base. recovered dearomatized is the wine dealcoholized reblending final by The alcohol. the remove osiunsi h oee tao ie(Belisario-S wine of ethanol concentration lowered the the the to in with leading removed constituents changes is volumetric which and vary- dioxide, ethanol, of sulfur presence of the concentrations to ing base, attributed were the wines of nonflavonoids dealcoholized in and and flavonoid phenolics, Differences esters, total wines. resveratrol, tartaric white flavonols, activities, and radical-scavenging red free the Spanish several under- with was process taken dealcoholization the from arising composition opnnscnb lae sn ceni lc”system. place” in “clean ancillary a and using column cleaned The be process. can sterilizing components or component a pasteurizing be a can and of conditions aseptic under operate to sealed rdc cuswt h ermtzdwn tasihl higher slightly of a pass at 38 wine 2nd (approximately dearomatized The volume. temperature the product with total occurs product of 1% approximately in h s aso ietruhteSCocr tlwtemperature low at occurs SCC the through 28 process.(approximately wine 2-stage of a pass of 1st in consists The technology concentration SCC alcohol using adjusting wine for finished process general The wine. affected. heat be to remain to unlikely SCC able are the are and flavors Since “fresh” delicate stage. temperatures, blending low the at at operate and wine can operation the processing to critical back a added before then wine from recovered or as be juice can equipment, from Flavors filtra- storage. as the during such oxidation operations of and winemaking fining, feature tion, certain in important lost be an can recovery aromas is The aromas concentrates. delicate juice grape of of of reduction production alcohol juice, and aromas, grape wines, delicate from of dioxide recovery sulphur including of industry, removal wine the in tions h C a enue oivsiaecniuu tao re- ethanol continuous investigate to used been has SCC The nivsiaino h fet fteSCo iephenolic wine on SCC the of effects the of investigation An C oei winemaking. in role SCC h C a nipratrl ntermvlo loo from alcohol of removal the in role important an has SCC The o.7,N.1 2012 1, Nr. 71, Vol. ◦ )advcu orcvrvltl iearomas wine volatile recover to vacuum and C) h C a ueosapplica- numerous has SCC The ◦ )advcu odtosto conditions vacuum and C) r ora fFo Science Food of Journal nhzand anchez ´ R37 h,

R: Concise Reviews in Food Science C, no toxic ◦ substances are required for use, it ishandled relatively (Rizvi inexpensive and and easily others 1994).dioxide in Furthermore, wine the production use does not ofis pose carbon ideally any suited legal difficulties for and extraction(Marignetti of and alcohol others from 1992). eitherof A wine alcohol or patented from wine beer process or for beer using the supercritical removal carbon dioxide ex- properties that can be exploited forThe separation use or of liquid extraction. carbon dioxideindustry for is supercritical gaining extraction popularity incritical and the temperature food offers for several this advantages gas as is relatively the low at 31 Vol. 71, Nr. 1, 2012 r Journal of Food Science Compression of a gas at temperatures above its critical point will Figure 11–Processing scheme for production of reduced alcohol wines using supercritical carbon dioxide extraction (Seidlitz and others 1991). R38 product discharge pump; 5: spinning10: cone recovered column; volatile 6: extract product pump. reinjection Courtesy pump; Flavourtech, 7: Lenehan product Rd. reinjection heater; 8: condensate cyclone; 9: vacuum pump; Figure 10–Typical layout of a spinning cone column and ancillary items. 1: product feed tank; 2: product feed pump; 3: product heat exchanger; 4: issue for finished wines, where fermentation has been completed. Supercritical solvent extraction Technologies for reduced alcoholic wines . . . and ethanol removal efficiency was foundIt to was be quite observed, high however, ataffected that 85%. the viability the of vacuum the yeastin applied cells, size and to and the the had cells became a SCC smaller different shape. This limitation should not be an result in formation of a supercritical fluid with increased solvent R: Concise Reviews in Food Science otnl sdfrtemnfcueo ag fhg-au foods. high-value of range a of manufacture the are for processes used extractive routinely plant, supercritical However, within of industry. technology this inflexibility wine of the uptake the the and for barriers distillation significant vacuum remain employed require- high commonly a for costs, a capital ment not High industry. is beverage the beverages within process alcohol pro- low the for of dioxide duction carbon using extraction supercritical feasible, opud VilyadLbes19) h hne nflavor in changes The 1998). Lubbers that these process and of aroma perception (Voilley a sensory of compounds thus wines, binding and volatility, in diminished the to materials increase leads proteinaceous also may to wine compounds the of from removal The ethanol 2000). techniques (Pickering aroma distillation processes thermal low-temperature of in than greater Loss be processing. to likely dealcoholized during is compounds from wines lost full-strength Thus, easily than 1998). more wines Lubbers be their and may (Voilley components to wines volatile due strength in ethanol solubility alcoholic increased of have full compounds presence these the and nature in nonpolar aroma reduced of volatility is The 1994). compounds Noble and acidity (Fischer oth- of astringency and acuity and Gawel decreases ethanol 2006; lowering Conversely, Pickering 2007). and ers (Nurgel others hotness and Gawel and 2005; 2007), and Pickering and (Fischer (Nurgel sweetness body 2006), 1994), Noble Pickering ethanol and and (Fischer Increased Nurgel bitterness alcohol. 1994; of Noble of perception the characteristics wine, enhance a sensory concentrations of weight the palate to have and sweetness will due the ethanol body upon of effect lack removal obvious wines The an 1993). alcoholic (d’Hauteville general reduced flavor A that and excep- juice. is GOX-treated the perception with from consumer article, manufactured this wines in of described tion by procedures produced the wines, of alcohol most reduced of evaluation sensory trolled esr ult fLwAchlWines Alcohol Low of Quality Sensory Mart provide and differences (Medina to considerable original presumably with the beverage from ethanol, a of produced some weight, of lack palate removal However, and cider. compounds original with aroma the product a to produced similarity components wine some base and Reblending aroma wine. the base of the aroma al- from of ethanol low extraction by followed producing supercritical compounds, for sequential using feasibility products technical cohol the shown have cider liquid a as bottom. away the drained from extracted component col- ethanol–water The the and from top. recovered head are the umn gas into dioxide carbon fed column and solvent mixture aromas the volatile of the alcohol bottom volatile which into the liquid in a and column as process pumped current is extraction counter dioxide) critical (carbon a The in 11. dioxide conducted Figure carbon is in supercritical the shown via illustrating is wine diagram extraction alcohol flow low A distil- of 1991). after production others remaining of and base portion (Seidlitz wine by aroma lation dearomatized returned the the is extracts scrubbing into dioxide bars sparging carbon 25 after to which distillate, fluid 18 the supercritical to the drop of expansion pressure at Partial by extraction pressure. bars supercritical 100 to to subjected 80 then and is alcohol containing compounds aroma fraction volatile captured The the distillation. high-vacuum subjects low-temperature beverages to beverage alcohol strength low alcoholic high of production the and traction . . . wines alcoholic reduced for Technologies eyltl ulse nomto saalbeta ecie con- describes that available is information published little Very io-cl xeiet o h rdcino dealcoholized of production the for experiments Pilot-scale ´ nz19) hl technically While 1997). ınez fehnlrmiigi h product. the quantity in the remaining upon ethanol dependent as- of is changes wine sensory dealcoholized with of sociated magnitude of perception The the characteristics. to compounds, mouthfeel changes aroma and interac- sweetness, of complex alcohol-related concentration of a loss and therefore volatility altered are removal of tion ethanol to due profile infiatvlm fpouti re obcm economically become to up- order a rapid in require product SCC, with an of process as volume osmosis such production significant Technologies, reverse producers. used that wine by widely ensured take a faults wine become some application has of of treatment flexibility in improvements for and Recent portability, of high. technology, relatively type membrane is this distillation equipment with low-temperature and associated plant costs of chemical distilla- development capital the The the thermal However, to processes. ethanol. with led associated of has compounds absence tion volatile the in and in to changes esters change due of chemical volatility alcohols from reduced higher arise and structure aromas may compound decreased wines volatile and alcohol mouthfeel reduced in in (d’Hauteville changes styles relative wine the these The of of 1993). sales one for is barriers products significant dealco- inferior most that organoleptically perception are wines consumer reality holized A commercial 1993). necessarily Young to not and publicity has (Howley marketing 1990s from early transcribed the associated been in risks consumption the alcohol of awareness with consumer increased from arising xlnto n rvso fifrainrgrigtespinning the regarding column. for information cone Delves Steve Trevor of provision and and and Griffith Code explanation Ltd., Standards Pty. Flavourtech Food from assistance Australian Sykes for of Corporation interpretation Brandy and in Wine Australian the from manufacturers. process many production wine for important an vintages remain between to likely consistency however, wines is, style products maintain in to concentration wine levels ethanol acceptable such to of for reduction future the unclear, the remains mar- Although been success. have with products keted strength several alcoholic challenges, these reduced of exacerbates commercial spite further In levels growth. sugar contaminant improve low potential and with composition associated weight varietal palate enhance grape to preserve with concentrates wines to juice ethanol-reduced order of in Blending conditions integrity. controlled product highly must in products these packaged highly and be growth a microbial creates beverage for product the product the from susceptible ethanol of of man- transport Removal by timely market. and faced to issue packaging real aseptic very is a ufactures wines, of strength production alcoholic for reduced employed technology the of Regardless feasible. uuePtnilo tao oie Wines Modified Ethanol of Potential Future nf nf.20b tnad451wn rdcinrqieet.Cner:Commonwealth Canberra: requirements. production containing wine 4.5.1 food Standard and 2002b. beverages Anzfa. alcoholic Anzfa of labelling 2.7.1 Standard 2002a. ANZFA. Washington, beverages—labeling. ANZFA malt and wine dealcoholized 510.400 during Sec. ethanol CPG of removal 2005. Partial Anon. 2010. J-M. Sablayrolles C, Camarasa A, Roy M, Bes E, Aguera References Acknowledgments fAsrla p. 6 Australia. of p. 6 Australia. of Commonwealth Canberra: alcohol. Administration. Drug and 61(1):53–60. Food D.C.: Vitic Enol J Am wines. reduced-alcohol obtain to fermentation h niiae nraei ae frdcdachlcbeverages alcoholic reduced of sales in increase anticipated The h uhr iht cnweg h dieo tpe Guy Stephen of advice the acknowledge to wish authors The o.7,N.1 2012 1, Nr. 71, Vol. r ora fFo Science Food of Journal R39

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