Hen egg white lysozyme is a hydrolytic enzyme effective at preventing the growth of Gram positive bacteria by degrading the bacterial cell wall to a point of cell lysis. Investigating lysozyme as a processing tool in wine to control the growth of lactic acid bacteria and malolactic fermentation has significant commercial interest. In this project, the interactions of lysozyme with wine components and wine was evaluated along with the efficacy of lysozyme in preventing malolactic fermentation (MLF) in Oregon Pinot Noir and Chardonnay. The information from this work, together with results from similar projects, will allow the development of guidelines for lysozyme use in commercial wine. Interactions of lysozyme with wine components were evaluated by measurement of enzymatic activity in the presence of wine acids, ethanol, and phenolics. Enzyme inhibition was observed, to various degrees, with all wine components. Crude grape tannin altered the availability of free enzyme by complexing to lysozyme and forming a precipitate. In a model wine system, lysozyme activity was reduced by 50% when tannin was present. Lysozyme addition to red wine resulted in a reduction in pigmented compounds and detectable sensory differences. Wine trials evaluated the efficacy of lysozyme in completely preventing malolactic fermentation (MLF) and terminating MLF midway through fermentation in Oregon Pinot Noir and Chardonnay. Vintages from 1993 and 1994 were treated without SO₂, with SO₂, with SO₂ plus a starter culture of Leuconostoc oenos. Each lot was divided into 0 ppm lysozyme (control), 250 ppm lysozyme, 500 ppm lysozyme, and 1000 ppm lysozyme. Lactic acid bacteria were enumerated monthly, for ten months. Lysozyme prevented malolactic fermentation in all wines at the treatment levels of 500 and 1000 ppm. In the 1993 Pinot Noir, 250 ppm lysozyme prevented MLF but only delayed MLF in the 1994 vintage. Lysozyme effectively terminated MLF at a concentration between 200 and 300 ppm in both Pinot Noir and Chardonnay. / Graduation date: 1996

Pinot noir has a reputation for lower color stability than other red wine varieties. Because it has relatively low anthocyanin and phenolic content and lacks acylated anthocyanin pigments compared to other red vinifera varieties, color extraction and stability are particularly important. Varying the processing during fermentation of red wine can produce high quality wines. Pectolytic enzymes are used in wine processing for many purposes from increasing juice yield and filtering rates to improving color and phenolic extraction. Macerating enzymes used in this study, Scottzyme Color Pro (Scott Laboratories), Scottzyme Color X (Scott Laboratories), Lallzyme EX (Lallemand), GB Rapidase EX Color (Gist Brocades), and Vinozyme G (Cellulo) were added to 1997 Oregon Pinot noir must prior to fermentation to observe color, aroma and flavor changes. These commercial enzymes have been reported to increase color and improve aroma and flavor of red wines. The effect of these enzymes had not previously been investigated on Oregon Pinot noir but the manufacturers reported increased polymeric phenols, polymeric anthocyanins, tannins, color stability, red hue and saturation of red wine varieties. These enzymes have varying manufacturer recommended usage levels and it is not known how the dosage levels and the enzymes themselves affect Oregon Pinot noir. Sensory evaluations of these wines, at a high and low dosage level, were conducted through free-choice profiling by winemakers and descriptive analysis from a trained panel. In addition, chemical analyses were performed and related to sensory panel results. Overall the addition of these enzymes to Oregon Pinot noir produced wines with greater purple, red descriptors and higher color intensity than the control wine from trained descriptive panel and winemaker panel results. In aroma, the enzyme treated wines were higher in vegetative and earthy descriptors compared to the control. GB Rapidase EX Color (Gist Brocades) was higher in bitterness flavor compared with other samples. Low enzyme wine treatments separated wine samples more from the control then high enzyme wine treatments. The color and appearance, aroma and flavor axes of the profile maps were not significant in the high enzyme treated wines as determined from the winemaker panel. Furthermore, the winemaker panel found acidity the only aroma or flavor descriptor significant in the high dosage ANOVA results. Whereas six descriptors in the low enzyme ANOVA results were significant in separating aroma and flavor samples. Except for Lallzyme EX (Lallemand) treated wine, the hunter colorimeter results showed all low enzyme treated wines were significantly (p<0.05) more red-purple (lower hues) than the high enzyme treated wines. / Graduation date: 1999

This research investigated the effects of Pediococcus spp. on Oregon Pinot noir wines. Pediococcus (P. parvulus (7), P. damnosus (1), P. inopinatus (1)) isolated from Oregon and Washington state wines demonstrated differences in their susceptibility to SO��� with some isolates growing well in model media at 0.4 mg/L molecular SO���. All isolates were all able to degrade p-coumaric acid to 4-vinyl phenol. The conversion of p-coumaric acid to 4-VP by pediococci resulted in accelerated production of 4-EP by B. bruxellensis in a model system. Growth of the pediococci isolates in Pinot noir wine resulted in a number of chemical and sensory changes occurring compared to the control. Very low concentrations of biogenic amines were measured in the wines with only wine inoculated with P. inopinatus OW-8 having greater than 5 mg/L. D-lactic acid production varied between isolates with OW-7 producing the highest concentration (264 mg/L). Diacetyl content of the wines also varied greatly. Some wines contained very low levels of diacetyl (< 0.5 mg/L) while others contained very high concentrations (> 15 mg/L) that were well above sensory threshold. Despite suggestions to the contrary in the literature, glycerol was not degraded by any of the isolates in this study. Color and polymeric pigment content of the wines also varied with wine inoculated with OW-7 containing 30% less polymeric pigment than the control. This may be related to acetaldehyde as a number of Pediococcus isolates, including OW-7, reduced the acetaldehyde content of the wine. Sensory analysis revealed differences in the aroma and mouthfeel of the wines compared to each other and to the control. In particular growth of some isolates produced wines with higher intensities of butter, plastic, and vegetal aromas while other also had lower perceived astringency. / Graduation date: 2013

Terroir is determined by a combination of factors in the vineyard including the grape varietal, geology and soil, soil hydrology, physiography, and climate. Although most studies have examined regional differences in wine flavors and associated provenance of wine based on chemistry, few have examined the chemistry of the soil and the ability to trace that chemistry to grape juice and, finally, to the wine. This dissertation examines what soil physical and chemical differences specific to this region might influence grape juice chemistry and wine chemistry. Wine-grapes in the Willamette Valley, Oregon, are grown on three major soil parent materials: volcanic, marine sediments, and loess/volcanic. Winemakers have observed differences in the flavor of Pinot Noir wine made from grapes grown on these different parent materials. This dissertation examines differences in the soil properties and elemental chemistry of the soil parent materials at various vineyards to document their effect on wine chemistry as a step towards understanding differences in flavor. All aspects of the terroir are controlled by carefully selecting vineyards with similar exposure and elevation, the same grape varietal and wine making techniques, and only the soils vary. The hypothesis is that the chemistry of the grape juice and wine reflect the soil in which the grapes were grown and that the three parent materials have soils that can be distinguished by their physical and chemical characteristics. Soil pits were excavated in 20 vineyards, soil properties were described in the field, and soil samples were later analyzed in the laboratory particle size, organic matter, color, pH, cation exchange capacity (ammonium acetate method), clay mineralogy (x-ray diffraction), and elemental chemistry (ICP-MS/AES). X-ray fluorescence was used to examine the pisolites. ICP-MS/AES was used for elemental analysis of grape juice and wines produced from these vineyards. Principal component analysis was used to compare soil physical and chemical characteristics, grape juice and wine chemistry. The physical characteristics of soils from all the three parent materials indicate: they are old (>50,000 years) based on their high clay content, low cation exchange capacity, red colors, and high Fe and Al content. These features indicate enough time has passed to reduce organic matter and other cations at depth, leave behind insoluble Fe and Al, and develop pedogenic clays. In my study region, volcanic and marine sediment soils are more developed with slightly lower acidity than the loess/volcanic soils. A new finding for this region is the presence of pisolites (Fe/Mg concretions) in the volcanic and the loess/volcanic soils, but absent in the marine sediment soils. Winemakers hypothesized that pisolites were present only in loess soils and influenced wine flavor in some way. Volcanic soils have the highest P, S, Fe, Co, Mn, and V concentrations and the lowest As and Sr values. Marine sediment soils have higher Cl and Sr and lower P, Co, Mn, Ba, and V concentrations than volcanic soils. Loess soils have the highest values of K and Mg and are similar to volcanic soils with higher P and V values and similar to marine sediment soils with higher Sr values. The main elements found to be significant in determining one parent material from another are V and Mn (volcanic soils), Mg and K (loess soils), and Sr (marine sediment or loess soils). Sr is slightly higher in grape juice and wine from vines grown on marine sediment parent material compared to volcanic and loess parent material, whereas Mn is higher in the juice and wine from grapes grown in volcanic parent material. P, S, Fe, Co, V, Cl, Ba, Mg, and K did not maintain their relative concentration levels from soil to grape juice to wine. The principal component analysis shows that soil and wine chemistry differs between parent material, but is inconclusive for grape juice chemistry.

Vineyard management strategies, including vineyard floor management and crop level management, can be used to influence vine vigor and fruit composition. Two studies were conducted to evaluate the impact of these practices on Pinot Noir in Oregon's Willamette Valley. Managing crop levels is common in cool climate vineyard production though it is a costly practice. With economic pressures, the premium winegrape industry is questioning whether they can reduce production costs and increase yields without compromising quality. A crop thinning trial was conducted in 2010 and 2011 to address these concerns and to better understand the role of vine balance on fruit composition. Crop levels were moderately (35% crop removed) or severely (65% crop removed) thinned at pre-bloom, fruit set, lag phase, or véraison and compared to full crop treatments. In both years, crop thinning reduced yields but had no effect on berry weight or cluster size. In 2010, poor fruit set reduced overall yields, and thinning treatments resulted in very few differences in vine growth, cluster architecture or fruit composition, including total soluble solids (TSS), pH, titratable acidity (TA), yeast assimilable nitrogen (YAN), anthocyanins, phenolics and tannins. In 2011, yields were much higher due to high fruit set and larger cluster size. No differences were found in vine growth (leaf areas or pruning weights) or fruit YAN, but thinned vines had higher TSS and pH and lower TA than full crop vines at harvest. Fruit thinned at lag phase and véraison had higher TSS and lower TA than fruit thinned pre-bloom. Intensity of thinning had a stronger influence on anthocyanin and tannin concentration than timing, while phenolics were not impacted by either factor. Ravaz index values (fruit yield/pruning weight) below 2.25 and leaf area to yield ratios of 2.25 to 3.25 m²/kg improved fruit composition in 2011 as did later season thinning, though data from the remaining years of this study will provide more insight into appropriate crop load metrics for cool climate Pinot Noir. A second study was implemented in 2011 to determine the impact of crop thinning in vines with different levels of vegetative vigor caused by three vineyard floor management techniques: permanent grass (Festuca rubra spp. rubra) cover (grass), alternating grass cover and tillage (grass & tilled), and tillage of every alleyway (tilled). Crop was thinned at the BB stage of berry development (EL stage 73) to one cluster per shoot (half crop) or not thinned (full crop); all cluster wings were removed at the time of thinning. Tillage treatments had been in place four years prior to the start of the study and competition for nitrogen in grass caused reduced early season vine growth, leaf chlorophyll and canopy size at both bloom and véraison while crop thinning increased canopy size at véraison. Yields were altered by tillage and crop thinning treatments, as grass had fewer clusters per shoot and berries per cluster, and crop thinning reduced yields to 64.7% of full crop across all tillage treatments. At harvest, grass had the lowest TA while TSS and pH were not affected by tillage. Crop thinning increased TSS but did not impact pH or TA. Anthocyanins were affected by both tillage and thinning and were found to be related to vine yield, YAN, leaf N, and leaf area index. Tannins were highest in grass but were not affected by crop thinning, and phenolics were not changed by either factor. Few interactions between tillage and crop thinning were found, but as variables such as yield per vine were impacted by both treatment factors, monitoring long term effects of crop / Graduation date: 2012

This research investigated yeast populations and diversity during pre-fermentation cold maceration and alcoholic fermentation of Vitis vinifera L. cv. Pinot noir grapes from a commercial vineyard (Dayton, OR). Fermentations were conducted at the Oregon State University research winery in 100 L tanks while grapes from the same vineyard lot were fermented at a commercial winery. Samples were taken daily during pre-fermentation maceration (9°C) and alcoholic fermentation (27°C) and plated on WL and lysine media to determine Saccharomyces and non-Saccharomyces populations and diversity. Total non-Saccharomyces populations increased from 1 x 10³ cfu/mL to 1 x 10⁵ cfu/mL during pre-fermentation cold maceration and reached a maximum of 1 x 10⁷ cfu/mL during alcoholic fermentation. Thirteen distinct yeast species were tentatively identified based on appearance on WL media and were initially screened for β-glucosidase activity using 4-methyllumbelliferyl-β-D-gluconopyranoside (4-MUG) plates. The identity of the isolates screening positive for β-glucosidase activity was determined by sequence analysis of the D1/D2 domain of the 26S rDNA gene. The five isolates identified were Metschnikowia pulcherrima, Hanseniaspora uvarum, Kluveromyces thermotolerans, and two Saccharomyces cerevisiae isolates. β-glucosidase activity was further characterized and quantified using a liquid media representing grape must conditions (pH 3.5, 20° Brix) at two temperatures (25°C and 8°C). While increasing sugar concentration suppressed the β-glucosidase activity of H. uvarum (-99%), β-glucosidase activity still remained relatively high for M. pulcherrima, S. cerevisiae isolate 1, and S. cerevisiae isolate 2. At 8°C, β-glucosidase activity was reduced for M. pulcherrima compared to activity at 25°C, but activity increased for K. thermotolerans, S. cerevisiae isolate 1, and S. cerevisiae isolate 2. The yeast isolates possessing β-glucosidase activity were used in fermentations of Vitis vinifera L. cv. Pinot Noir grapes. The grapes were treated with high hydrostatic pressure (HHP) to inactivate naturally occurring yeast and bacteria. All yeast isolates grew during pre-fermentation cold maceration (7 days at 9°C) and populations increased 3 to 4 logs. Following pre-fermentation cold maceration, all ferments were warmed to 27°C and inoculated with S. cerevisiae RC212. Alcoholic fermentations were all complete within eight days and after pressing wines were analyzed for volatile aroma compounds by SPME-GC-MS. The presence of different yeast isolates during pre-fermentation cold maceration resulted in wines with unique aroma profiles. Ethyl ester concentrations were highest in the wine that did not undergo a pre-fermentation cold maceration, while concentrations of branch-chained esters were higher in the treatments with yeast present during pre-fermentation cold maceration. Pre-fermentation cold maceration with yeast isolates demonstrating β-glucosidase did not affect the concentration of β-damascenone or β-ionone. Wines that had undergone pre-fermentation cold maceration with S. cerevisiae isolate 1, S. cerevisiae isolate 2, and a combination of all isolates resulted in over twice the concentration of β-citronellol over wines that did not undergo a pre-fermentation cold maceration. / Graduation date: 2013

Graduation date: 2004