Thesis (MScAgric)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: The distinctive varietal flavour of wines is a combination of absolute and relative
concentrations of chemical compounds. Volatile compounds are responsible for the
odour of wine and non-volatiles cause the sensation of flavour. Accompanying these
senses, a third, tactile, sense of ‘mouth-feel’ is recognizable. This forms the complete
organoleptic quality of wine.
Several hundred different compounds are simultaneously responsible for the
odour release in wine, and since there is no real character impact compound, the
aroma of wine can be described as a delicate balance of all these compounds. One
of the most important groups of volatiles is the monoterpenes, which play a role in
both aroma and flavour. This is especially significant for the Muscat varieties, but
these flavour compounds are also present in other non-muscat grape varieties,
where they supplement the varietal aroma. Monoterpenes occur in wine as free,
volatile and odorous molecules, as well as flavourless non-volatile glycosidic
complexes. The latter slowly releases monoterpenes by acidic hydrolysis, but the
impact on varietal aroma is considered insufficient for wines that are consumed
young. It is therefore important to supplement the release mechanism, in order to
enhance the varietal aroma of the wine. The enzymatic hydrolysis mechanism
functions in two successive steps: firstly, depending on the precursor, the glycosidic
linkage is cleaved by α-L-arabinofuranosidase, α-L-rhamnosidase, β-D-xylosidase or
β-D-apiosidase. The second step involves the liberation of the monoterpene alcohol
by a β-glucosidase. This enzymatic hydrolysis does not influence the intrinsic
aromatic characteristics of the wine, as opposed to acid hydrolysis.
Pectolytic enzymes play an important role in cell elongation, softening of tissue
and decomposition of plant material. These enzymes are used to improve juice
yields, release colour and flavour compounds from grape skins, as well as improve
clarification and filterability. Pectolytic enzymes work synergistically to break down
pectins in wine. Protopectinase produce water-soluble and highly polymerised pectin
substances from protopectin, it acts on non-methylated galacturonic acid units. Pectin
methylesterase split methyl ester groups from the polygalacturonic chain.
Polygalacturonase break down the glycosidic links between galacturonic acid units.
Pectin and pectate lyases have a β-eliminative attack on the chain and it results in
the formation of a double bond between C4 and C5 in the terminal residues.
From the above it can be seen that enzymes play a pivotal role in the
winemaking process. Unfortunately, in winemaking a lot of factors can influence the
effects of enzymes. One possible factor in the wine medium is the presence of acidprotease,
from yeast and/or fungal origin. This type of enzyme utilizes other enzymes
as substrates and renders them useless. Pure enzyme preparations were used to
study the interactions of a yeast acid-protease and a report activity (β-glucosidase) in
vitro. A bottled wine and a buffer were used as in vitro conditions. Enzyme assays were performed to determine the relative activity over a number of days. The results
indicated that even though both enzymes showed activity in both the media, the
yeast protease did not have any significantly affect on the report activity.
Subsequently wine was made from Sauvignon blanc grapes, with varying enzyme
preparation additions. Enzyme assays were performed during the fermentation; and
chemical, as well as sensory analysis were done on the stabilized wine. The results
confirmed that the yeast protease did not have any significant affect on the report
activity in these conditions. The protease’s inability to affect the report activity seems
unlikely due to the fact that it is active at a low pH range and has been suggested as
the only protease to survive the fermentation process. It seems possible that a winerelated
factor, possibly ethanol, is responsible. Thus it seems that yeast protease
does not threaten the use of commercial enzymes in the winemaking process in any
significant way.
Future work would entail more detailed enzyme studies of interactions
between protease, both from yeast and fungal origin, and other report activities in
specified conditions. The degradation capability could be directed towards unwanted
enzyme activities that cause oxidation and browning of the must. The
characterization of interactions between protease and β-glucosidase activities may
hold key to producing wines with enhanced aroma and colour potential, as well as
the elimination of unwanted enzyme activities. / AFRIKAANSE OPSOMMING: Die herkenbare kultivar karakter van wyn is ‘n kombinasie van absolute en relatiewe
konsentrasies van verskeie chemiese komponente. Vlugtige komponente is
verantwoordelik vir die geur, of aroma, van wyn en die nie-vlugtige komponente
veroorsaak die sensasie van smaak. ‘n Derde, fisiese sensasie, die ‘mondgevoel’, is
ook herkenbaar. Dit vorm die omvattende organoleptiese kwaliteit van die wyn.
‘n Paar honderd verskillende komponente is gelyktydig verantwoordelik vir die
aroma vrystelling in wyn en omdat daar geen werklike karakter ‘impak’ komponent is
nie, kan die aroma van wyn beskryf word as ‘n delikate balans van al die betrokke
komponente. Een van die mees belangrike groepe vlugtige komponente is die
monoterpene wat ‘n rol speel in beide aroma en smaak. Dit is veral belangrik by
Muskaat kultivars, maar hierdie aroma komponente is ook teenwoordig in niemuskaat
druif kultivars, waar hulle bydra tot die kultivar karakter en aroma.
Monoterpene kom in wyn voor as vry, vlugtige en aromatiese molekules en in
geurlose, nie-vlugtige glikosidies-gebonde komplekse. Die gebonde vorm word stadig
vrygestel deur ‘n suurhidrolise, maar dit word as onvoldoende beskou vir wyne wat
vroeg gedrink word. Dit is dus belangrik dat die vrystelling van geurstowwe verhoog
word om die kultivar karakter van die wyn te versterk. Die ensiematiese hidrolise
proses behels twee opeenvolgende stappe: eerstens, afhangende van die aard van
die voorloper, word die glikosidiese verbinding deur α-L-arabinofuranosidase, α-Lramnosidase,
β-D-xilosidase, of β-D-apiosidase gebreek. In die tweede stap word die
monoterpeen-alkohol deur β-glukosidase vrygestel. Hierdie ensiematiese afbraak
proses verander nie die intrinsieke aromatiese kenmerke van die wyn, soos met
suurhidrolise die geval is nie.
Pektolitiese ensieme speel ‘n fundamentele rol in selverlenging, sagwording en
afbraak van plant materiaal. Hierdie ensieme word gebruik om sap opbrengs te
verhoog, aroma en smaak komponente vry te stel uit die doppe, asook om
sapverheldering en filtrasie te verbeter. Die pektolitiese ensieme werk op ‘n
sinergistiese wyse om pektien in wyn af te breek. Protopektinase produseer wateroplosbare
en hoogs gepolimeriseerde pektien uit protopektien, slegs uit niegemetileerde
galakturoonsuur eenhede. Pektien metielesterase verwyder metielester
groepe van die poligalakturoonsuurketting. Die glikosidiese bindings tussen
galakturoonsuur eenhede word deur poligalakturonase afgebreek. Pektien- en
pektaat-liase het ‘n β-eliminasie aanslag op die ketting en as gevolg daarvan word
dubbelbindings tussen C4 en C5 in die terminale residue gevorm.
Vanuit bogenoemde is dit dus duidelik dat ensieme ‘n kardinale rol speel in die
wynbereidingsproses. Ongelukkig is daar ‘n verskeidenhied van faktore wat die
werking van ensieme in die wynbereidingsproses kan beïnvloed. Een moontlike
faktor is die teenwoordigheid van ‘n suur-protease, van fungisidiese en/of gis
oorsprong, in die wynmedium, omdat dit ander ensieme as substraat kan benut en degradeer. Suiwer ensiem preparate is gebruik om die ensiem interaksie tussen ‘n
gis suur-protease en ‘n verslag aktiwiteit (β-glukosidase) in vitro te ondersoek. ‘n
Gebotteleerde wyn en ‘n buffer is gebruik om die in vitro kondisies na te boots.
Relatiewe ensiem aktiwiteit is ontleed oor ‘n aantal dae. Beide die ensieme het
aktiwiteit getoon in die media, maar gis protease het geen statisties beduidende
invloed gehad op die aktiwiteit van die verslag ensiem nie. Daaropvolgend is wyn
berei van Sauvignon blanc druiwe, met verskillende ensiempreparaat toevoegings.
Die ensiemaktiwiteit is deurlopend tydens fermentasie gemeet. Na afloop van
stabilisasie is chemiese, sowel as sensoriese ontledings op die wyn gedoen. Die
resultate het bevestig dat gis protease, onder hierdie kondisies, geen beduidende
invloed op die verslag aktiwiteit gehad het nie. Die protease se onvermoë om die
verslag aktiwiteit beduidend te beinvloed blyk onwaarskynlik aangesien die suurprotease
aktief is by lae pH vlakke en dit as die enigste protease voorgestel is wat
die fermentasie proses kan oorleef. Dit blyk asof ‘n wyn-verwante faktor, moontlik
etanol, hiervoor verantwoordelik kan wees. Dus hou protease geen gevaar in vir die
gebruik van kommersiële ensieme in wynbereiding nie.
Navorsing kan in die toekoms fokus op meer gedetailleerde ensiem interaksie
studies tussen protease en ander ensiem aktiwiteite, in gespesifiseerde kondisies.
Die degradasie kapasiteit kan moontlik aangewend word om ongewenste ensiem
aktiwiteite, wat byvoorbeeld oksidasie en verbruining veroorsaak, te verminder. Die
karakterisering van die interaksies tussen protease en β-glukosidase kan dus die
sleutel wees tot die produksie van wyne met verhoogde aroma potensiaal, asook die
eliminasie van ongewenste ensiematiese aktiwiteite.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:sun/oai:scholar.sun.ac.za:10019.1/16445 |
| Date | 12 1900 |
| Creators | Swart, Elsa Marita |
| Contributors | Van Rensburg, Pierre, Otero, Ricardo Codero, University of Stellenbosch. Faculty of Agrisciences. Dept. of Viticulture and Oenology. |
| Publisher | Stellenbosch : University of Stellenbosch |
| Source Sets | South African National ETD Portal |
| Language | en_ZA |
| Detected Language | Unknown |
| Type | Thesis |
| Format | xii, 94 leaves : ill. |
| Rights | University of Stellenbosch |
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