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Well-Fed and Heathly Wine Yeast

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Well-Fed and Heathly Wine Yeast YEAST NUTRITION AND PROTECTION FOR RELIABLE ALCOHOLIC FERMENTATION THE STATE OF THE ART Natural solutions that add value to the world of winemaking THE STATE OF THE ART YEAST NUTRitiON AND PROTECtiON FOR RELIABLE ALCOHOLIC FERMENTAtiON LALLEMAND, A LEADING PRODUCER OF OENOLOGICAL YEASTS AND BACTERIA AND THEIR FERMENTAtiON NUTRIENTS, IS A PRIVATELY OWNED CANADIAN CORPORAtiON. THE OENOLOGY GROUP, BASED IN TOULOUSE (FRANCE), IS FOCUSED ON RESEARCH AND DEVELOPMENT, BOTH IN-HOUSE AND IN COLLABORAtiON WitH RENOWNED RESEARCH INStitUTES. THE PURPOSE OF THIS DOCUMENT IS TO PROVIDE WINEMAKERS AND OENOLOGISTS WitH A DESCRIPtiON OF THE CURRENT SCIENtiFIC UNDERSTANDING ON YEAST NUTRitiON AND PROTECtiON FOR RELIABLE ALCOHOLIC FERMENTAtiON MANAGEMENT. Document prepared by Anne Julien (Lallemand SAS, France), Didier Theodore (Lallemand SAS, France), Gordon Specht (Lallemand, North America), Ann Dumont (Lallemand Inc., Canada) and Cecile Brung (Lallemand SAS, France). ACKNOWLEDGEMENTS: Dr. Charles Edwards - Washington State University (USA) Pr. Jean-Marie Sablayrolles - UMR Sciences pour l’Œnologie - INRA - Montpellier (France) Dr. Jean-Michel Salmon - UMR Sciences pour l’Œnologie - INRA - Montpellier (France) I INTRODUCTION ..................................... 04 III YEAST NUTRITION MANAGEMENT .......... 24 I.1 Nitrogen ............................................. 06 III.1 The importance of Complex nutrients such as Fermaid® .............................. 26 I.2 Oxygen/survival factors (sterols) ............ 08 I.3 Minerals ................................................. 10 I.4 Vitamins ................................................... 12 IV 4 STUCK FERMENTATION MANAGEMENT ................ 30 IV.1 Prepare the stuck wine .......................................... 32 II SELECTION AND PROPER HANDLING OF YEAST .......... 14 IV.2 Prepare the rescue yeast ........................................... 32 ® II.1 YSEO yeast .............................................................. 16 IV.3 Activate the prepared rescue yeast II.2 Specific selected Yeast (characterization with nutrients and sugar ........................................................... 33 of Nitrogen requirements) ........................................................ 18 IV.4 Restart the fermentation and sequential II.3 Yeast Rehydration optimization: NATSTEP® process .............. 20 addition of the stuck wine ................................................................. 33 2 3 THE STATE OF THE ART 1. INTRODUCtiON TO ASSURE A COMPLETE AND REGULAR FERMENTAtiON, it IS NECESSARY FOR THE GRAPE MUST TO HAVE A REASONABLE BALANCE OF NUTRitiONAL, PHYSICAL AND CHEMICAL ENVIRONMENT THAT WILL ALLOW THE YEAST TO MULtiPLY AND THRIVE. IT IS ESSENtiAL TO HAVE AN OPtiMUM DEVELOPMENT OF HEALTHY YEAST CELLS IN ORDER TO REDUCE THE RISK OF SLUGGISH AND STUCK FERMENTAtiONS. THE FOLLOWING FACTORS ARE THE MOST LIKELY CAUSES OF STUCK AND SLUGGISH FERMENTAtiONS: N ITROGEN DEFICIENCY LACK OF OXYGEN LACK OF TEMPERATURE MANAGEMENT IMPROPER YEAST REHYDRATION AND HANDLING LACK OF GROWTH FACTORS SUCH AS VITAMINS AND MINERALS HIGHLY CLARIFIED MUSTS INHIBITORY METABOLITES PRODUCED BY YEASTS EXCESSIVE SULPHUR DIOXIDE OR AGROCHEMICAL RESIDUES I.1 Nitrogen ......................................... 06 SOME VitiCULTURAL FACTORS CAN AFFECT THE CONDitiON OF MUST FERMENTAtiON. AMONG THESE FACTORS, IT IS IMPORTANT TO REMEMBER: THE HIGH DEGREE OF MATURitY RESULtiNG IN A HIGH CONCENTRAtiON OF FERMENTABLE SUGARS IN THE GRAPE MUST IS ASSOCIATED WitH LIMitED LEVELS OF AVAILABLE NitROGEN 2 Oxygen/survival factors (sterols) .............. 08 INCREASING THE RISK OF SLOW OR STUCK FERMENTATIONS. I. A POOR SANITARY CONDITION OF THE GRAPES RESULTING IN THE PRESENCE OF LARGE POPULAtiONS OF UNDESIRABLE MICROORGANISMS (BACTERIA AND YEASTS, BOTRYtiS) REDUCES THE AVAILABILitY OF NitROGEN, PREVENtiNG DESIRED YEAST GROWTH AND THEIR ACTIVITY. 3 Minerals ............................................................ 10 AGROCHEMICAL RESIDUES ARE ALSO IMPLICATED IN FERMENTAtiON ISSUES. SOME OF I. THEM ARE RESPONSIBLE FOR A LONGER LAG PHASE DELAYING THE START OF ALCOHOLIC FERMENTATION (SUAREZ ET AL., 2000). THESE FACTORS ARE USED TO ASSESS THE POTENtiAL RISK OF STUCK FERMENTAtiON, Vitamins .......................................................................... 12 BUT WHEN A PARTICULAR WINE STYLE IS DESIRED, THE INTERVENTION IN THE VINEYARD IS I.4 NOT ALWAYS POSSIBLE. HOWEVER, WINEMAKERS CAN COMPENSATE THE SITUATION BY BEING PROACTIVE DURING FERMENTATIONS. 4 5 THE STATE OF THE ART I.1 NitROGEN NitROGEN, THE MOST IMPORTANT YEAST NUTRIENT, IS A KEY FACTOR THAT HAS A SIGNI- FICANT IMPACT ON WINE FERMENTAtiON. THIS NUTRIENT INFLUENCES BOTH FERMENTAtiON KINEtiCS AND THE RESULtiNG WINE QUALitY (AGENBACH, 1977; BEZENGER AND NAVARRO, 1987). IN GENERAL, NitROGEN MUST DEFICIENCY LIMitS YEAST GROWTH AND FERMENTAtiON SPEED (BELY ET AL., 1990). itrogen content in grape juice is variable and amino acids. The yeast assimilable nitrogen (YAN) is ranges from 60 to 500 mg nitrogen/L (J.M composed of ammonium and alpha amino acids NSablayrolles and J.M Salmon, 1996) depen- except proline which is not assimilable by yeast. NitROGEN IMPACT ON ALCOHOLIC FERMENTAtiON HOW MUCH YAN IS NEEDED? ding on grape variety, vintage and microclimate. Sometimes, YAN levels in must are not measured YAN content has the most influence on the fermen- The minimum quantity of YAN is 150 to Grape musts contain nitrogen under different forms: even though they are frequently deficient. tation speed; it impacts the yeast biomass at the 200mg/L and lower levels in the must are consi- + proteins, peptides, ammonium ions (NH4 ) and alpha beginning of fermentation, as well as the sugar dered as nitrogen deficient. This minimal level NITROGEN ASSIMILATION BY YEAST transport kinetics during fermentation. As soon as a varies based on different parameters: Chardonnay 220 g/L, fermented with EC 1118 (20 g/hL) Nitrogen is essential for yeast protein synthesis must is nitrogen depleted at the end of the growth Temperature: an increase in temperature stimu- 1.4 20 mgN/L throughout fermentation. However, the different phase, there is a decrease in protein synthesis and lates the growth of yeast and fermentation is Ammonium (Inorganic YAN) types of nitrogen are assimilated all differently. sugar transport activity (Bely et al., 1994). YAN addi- faster requiring increased amounts of nitrogen. 1.2 Organic nitrogen in the form of amino acids enters tion to nitrogen deficient must leads to a significant Oxygen: when adding more O2 to the must, 1.0 the cell through an active transport and can be accu- decrease in fermentation length. After a YAN addi- nitrogen is captured faster and more is needed in 0.8 mulated inside the yeast cells (Bisson, 1991). The tion, there is a reactivation of the protein synthesis comparison to fermentations taking place under main system is the GAP (General Amino Permease) and the sugar transport speed, which results in an anaerobic conditions (white wine). 0.6 40º Slope which is able to transport all alpha amino acids. increase in the fermentation rate. Initial sugar content: the higher the initial concen- 0.4 5 mgN/L Different groups have been determined as a function tration, the more YAN is required. ␣-AMINO N 8º Slope 0.2 (Organic YAN) of their assimilation order in a must from “very fast” NitROGEN AND AROMAtiC PROFILE OF WINE Turbidity: when musts are over clarified, many Fermentation Speed (g/L.h) Fermentation to ”partial“ (Henschke et al., 1993). Once inside the Amino acids composition of the must is quite important nutritional factors for yeast are removed, there- 0 0 50 100 150 200 yeast cell, these alpha amino acids can be integrated for the wine’s aromatic profile. Higher alcohols and fore it is necessary to supplement them with the Time (hr) directly into proteins, degraded into ammonium esters (ester acetates and ethyl esters) production is addition of complete and balanced nutrients. ϳ 46 hr fermentation time difference + (NH4 ), or converted into glutamate. Their assimila- influenced by the amino acids must content. Sulfur Efficiency of organic versus inorganic nitrogen on EC1118 wine tion by the yeast is more gradual and efficient during compounds production is also linked with the nitrogen yeast fermentation kinetics as measured by CO2 release. The fermentation compared to inorganic nitrogen. metabolism. When assimilable nitrogen is depleted and/ sluggish fermentation with inorganic nitrogen added had 4 Ammonium nitrogen, the other type of assimilable or deficiency in methionine and its derivates appears, times the YAN level than the organic nitrogen addition which nitrogen referred to as inorganic nitrogen, is rapidly H2S synthesis is favoured. finished alcohol fermentation ~46 hours earlier. consumed by yeast but is less beneficial. 6 7 THE STATE OF THE ART I.2 OXYGEN /SURV IVAL FACTORS (STEROLS) OXYGEN PLAYS AN ESSENtiAL ROLE IN ALCOHOLIC FERMENTAtiON HELPING THE DEVELOP MENT OF AN ADEQUATE YEAST POPULAtiON AND MAINTAINING THEIR VitALitY. OXYGEN IS REQUIRED FOR THE SYNTHESIS OF SURVIVAL FACTORS SUCH AS STEROLS AND UNSATURATED FAttY ACIDS (KIRSOP, 1973; KIRSOP, 1978; ALEXANDRE ET AL., 1994; FORNAIRON- BONNEFOND ET AL., 2002), WHICH ARE COMPONENTS OF THE YEAST CELL MEMBRANE. THEY PLAY A KEY ROLE ON THE MEMBRANE STRUCTURE HELPING TO MAINTAIN THE MEMBRANE FLUIDitY, CELL INTEGRitY AND VIABILitY. OXYGEN AND STUCK FERMENTATIONS During the yeast growth phase, each multiplication OXYGEN ADDITION Oxygen can be incorporated efficiently into the must
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