Ciencia y Tecnología Alimentaria ISSN: 1135-8122 [email protected] Sociedad Mexicana de Nutrición y Tecnología de Alimentos México Rogerson, F.S.S; Vale, E.; Grande, H.J.; Silva, M.C.M. Alternative processing of port-wine using pectolytic enzymes Ciencia y Tecnología Alimentaria, vol. 2, núm. 5, julio, 2000, pp. 222-227 Sociedad Mexicana de Nutrición y Tecnología de Alimentos Reynosa, México Available in: http://www.redalyc.org/articulo.oa?id=72420501 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative Cienc. Tecnol. Aliment. Vol. 2, No. 5, pp. 222-227, 2000 Copyright 2000 Asociación de Licenciados en Ciencia y Tecnología de los Alimentos de Galicia (ALTAGA). ISSN 1135-8122 ALTERNATIVE PROCESSING OF PORT-WINE USING PECTOLYTIC ENZYMES PROCESADO ALTERNATIVO DEL VINO DE OPORTO USANDO ENZIMAS PECTOLÍTICOS PROCESADO ALTERNATIVO DO VIÑO DE OPORTO USANDO ENZIMAS PECTOLÍTICOS Rogerson, F.S.S*1; Vale, E.3; Grande, H.J.2; Silva, M.C.M.3 1* Departmento de Química, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre 687, 4169-007 Porto. Portugal. E-Mail: [email protected] 2 Licentec, Bernadottelaan 15, P.O.Box 8323, 3503 RH Utrecht, The Netherlands. 3 Escola Superior de Biotecnologia/Universidade Católica. Rua Dr.Antonio Bernardino de Almeida, 4200 Porto. Portugal. Recibido: 24 de Octubre de 1999; recibida versión revisada: 9 de Marzo de 2000; aceptado: 15 de Marzo de 2000 Received: 24 October 1999; rreceived in revised form: 9 March 2000; accepted: 15 March 2000 Abstract The objective of the present study was to investigate the application of a commercial pectolytic enzyme preparation “Ultrazym” during grape maceration for the alternative processing of 7 single varietal Port-Wines (Tinta Barroca, Mourisco Tinto, Tinta Roriz, Rufete, Tinta da Barca, Tinta Santarém and Touriga Nacional). The effect of enzymatic treatment resulted in improvements in: i) juice yield; ii) colour extraction; iii) wine filterability. Juice yields increased by up to 6%, wine colour intensified by up to 40%, and wine filtration speed was considerably more rapid in the enzyme treated wines. Ó 2000 Altaga. All rights reserved. Keywords: Wine, pectolytic enzyme. Resumen El objetivo de este trabajo fue investigar la aplicación de un preparado enzimático comercial durante la maceración de la uva para realizar un procesado alternativo de 7 variedades monovarietales de vino de Oporto (Tinta Barroca, Mourisco Tinto, Tinta Roriz, Rufete, Tinta da Barca, Tinta Santarém y Touriga Nacional). El efecto del tratamiento enzimático resultó en una mejora: i) del rendimiento en jugo; ii) de la extracción de colorantes; iii) filtrabilidad del vino. El Rendimiento en jugo se incrementó hasta un 6%, la intensidad del color del vino hasta un 40% y la velocidad de filtración fue considerablemente más rápida en los vinos preparados con enzimas. Ó 2000 Altaga. Todos los derechos reservados. Palabras clave: Vino, enzimas pectolíticos. Resumo O obxetivo deste traballo foi investiga-la aplicación dun preparado enzimático comercial durante a maceración da uva para realizar un procesado alternativo de 7 variedades monovarietais de viño de Porto (Tinta Barroca, Mourisco Tinto, Tinta Roriz, Rufete, Tinta da Barca, Tinta Santarém e Touriga Nacional). O efecto do tratamento enzimático resultóu nunha mellora: i) do rendemento en xugo; ii) da extracción de colorantes; iii) filtrabilidade do viño. O Rendemento en xugo incrementóu-se hasta un 6%, a intensidade do color do viño hasta un 40% e a velocidadd de filtración foi considerablemente más rápida nos viños preparados con enzimas. Ó 2000 Altaga. Todolos dereitos reservados. Palabras chave: Viño, enzimas pectolíticos. 222 ALTAGA ©2000 Rogerson et al.: Alternative processing of Port-wine using pectolytic enzymes INTRODUCTION stored for 9 months at -20oC, were then defrosted, destemmed, weighed and divided. The first half (blank) Over the past few decades, increasing interest has and second half (added “Ultrazym”, 30mg/L) were crushed been shown in the application of enzymes in wine-making. with the addition of SO2 (30mg/L). The crushed grapes, Enzymatic treatments of grapes, musts and wine have contained in 2.5L glass flasks, were left macerating for 8 multiple purposes, since they influence wine clarification hours (20oC), after which the Saccharomyces cerevisiae and filtration, juice yield, colour and aroma extraction and yeast, Firmivin, was inoculated (200mg/L). At specific wine stability (Ough et al., 1975; Villettaz, 1984; Zamorani, gravity (S.G.) 1.045, the fermenting mash was pressed 1989; Canal-Llauberes, 1989 &1990; Villettaz and (cone press) and the resulting juice was filtered (Whatman Dubourdieu, 1991). Nº1 filter paper) and yield determined. The semi-fermented must was then fortified with alcohol (99.8% v/v, Merck) Industrial pectolytic enzyme preparations have to give calculated alcoholic strength of 20% v/v. The been widely used for many years, most being derived from resulting Port-Wines were bottled and allowed to mature cultures of Aspergillus niger which is a species accepted for two months. The micro-vinification procedure is as G.R.A.S. (Generally Recognised As Safe) (Canal- represented in Figure 1. Llauberes, 1993). Besides the main pectolytic activities (pectinmethylesterase, polygalacturonase, pectin lyase & Analytical methods pectate lyase), industrial pectinase preparations also contain hemicellulolytic, cellulolytic and side-activities Juice volume including glycosidases (Cordonnier et al., 1989) and Port-Wine juice volume was measured before the proteases (Schmitt et al., 1989). addition of the fortification alcohol, but after the removal of both the skins (pressing) and insoluble solids (filtration). Pectolytic enzyme preparations are valuable processing tools for many wine and grape processing Colour applications, including: mash treatment for juice extraction Colour measurements were made after 2 months (Ough & Crowell, 1979; Canal-Llauberes, 1989); juice wine maturation, using a Pye-Unicam spectrophotometer clarification and wine filtration (Brown & Ough, 1981; according to the method of Jackson et al. (1978). Canal-Llauberes, 1989); colour extraction (Ough et al., 1975; Castino & Ubigli, 1979) and aroma liberation Measurements included: Wine colour (W.C.) at (Cordonnier et al., 1989; Rogerson et al., 1995). wine pH (l520nm, 1mm) and total pigment colour (W.C.A.) measured in 1.0M HCl (1% solution, l520nm, Recent investigations (Birks & Sarmento, 1991; 10mm). Bakker et al., 1996) compared the two most typically used maceration techniques for Port-Wine production (open tank Total Phenolics treading & static tanks with pumped over juice), however Total phenolics were measured in 1.0M HCl (1% nothing appears to have been studied, looking at the solution, l280nm, 10mm), and calculated according to the application of pectolytic enzymes for alternative processing method of Somers & Evans (1977) using the formulae of Port grapes, musts and wines. Therefore the objective (A280nm –4). of this work was to investigate the influence of a pectolytic enzyme preparation on juice yield, colour release and wine filterability, during the production of several single varietal Port-Wines. GRAPES MATERIALS AND METHODS Defrost Destem 1/2 1/2 Crush Crush + SO (30 mg/L) The A. niger enzyme preparation was “Ultrazym 2 + Ultrazym (30mg/L) + SO2 (30mg/L) 100G” from Novo Nordisk. The inoculation yeast was Maceration Maceration o Saccharomyces cerevisiae , “Firmivin”, from Gist- 8 Hrs, 20 o C 8 Hrs, 20 C Brocades. Yeast 200mg/L Yeast 200mg/L Fermentation 25 º C Fermentation 25 º C until S.G. 1.045 until S.G. 1.045 The grape cultivars (Touriga Nacional, Tinta Press, Filter Press, Filter Roriz, Tinta Barroca, Mourisco Tinto, Tinta da Barca, & Fortify & Fortify Bottle & Bottle & Tinta Santarém and Rufete; 1992 vintage) were from Tua Storage Storage (20 º C) (20 º C) in the Douro valley (Portugal). All reagents used were o research grade. PORT WINE PORT WINE (BLANK) (Ultrazym) Red Grape Micro-Vinifications The grapes, picked at commercial maturity, were Figure 1. Processes of micro-vinifications performed in this study (S.G. taken back to the University in Porto where they were = Specific Gravity) . 223 Cienc. Tecnol. Aliment. Vol. 2, No. 5, pp. 222-227, 2000 ISSN 1135-8122 ©2000 ALTAGA Cross-Flow Micro-Filtration Maceration gives red wine its essential colour and tannic Micro-filtration analysis was performed on the structure, being particularly important for grape processing fortified wines after two months bottle storage, using a for the production of great red wines intended for ageing. laboratory scale Carbosep® 40 (TECH-SEP, Grupe Rhône- Apart from traditional maceration, alternative extraction Poulenc), equipped with a tubular membrane (zirconion technologies include thermovinification (Dubourdieu, oxide contained in a carbon support) with pore size 0.14mm 1986) and enzymic methods (Villettaz and Dubourdieu, and filtration area of 0.008m2. The membrane was initially 1991). washed for 10 minutes with wine which was discarded. Each filtration was carried out using 120mL of wine, using Thermovinification (typically 70-80ºC) has been a transmembranal pressure of 1.7 bar. The filtration shown as beneficial for increasing colour (anthocyanin equipment was allowed to filter the wine, using