Non-alcoholic beverage

Abstract

The present invention relates to a process of producing a non-alcoholic beverage, in particular a fermented, non-alcoholic beverage which is a wine analogue.

Description

[0001] NON-ALCOHOLIC BEVERAGE

[0002] TECHNICAL FIELD OF THE INVENTION

[0003] The present invention relates to a process of producing a non-alcoholic beverage, in particular a non-alcoholic beverage which is a wine analogue.

[0004] BACKGROUND OF THE INVENTION

[0005] Consumers are increasingly looking for non-alcoholic alternatives to traditionally alcoholic beverages. In the case of wine, the conventional method of producing nonalcoholic wine involves producing a traditionally fermented wine, and later de- alcoholising that wine by use of e.g. a spinning cone. The de-alcoholization process also removes some of the flavour profile, which is typically compensated for by adding additional flavouring and / or sugar. Though being conventional, such de-alcoholization process is time and energy consuming.

[0006] Thus, there remains a need for processes for providing non-alcoholic alternatives to traditional fermented alcoholic beverages which are more efficient and environmentally sustainable. There remains also a need for such beverages which have improved flavour profile.

[0007] SUMMARY OF THE INVENTION

[0008] The invention relates to a process for producing a non-alcoholic beverage comprising the steps of a. Providing a starting material comprising grape extract; b. Adding at least one ethanol production inhibitor, c. Inoculating the starting material with at least one malolactic bacterial strain; d. Fermenting the starting material including the at least one malolactic bacterial strain, thereby achieving a non-alcoholic beverage having an alcohol content of not more than 0.5 % Alcohol By Volume (ABV).

[0009] One advantage of the invention may be that an improved non-alcoholic grape extract based beverage may be obtained with improved organoleptic properties, e.g. improved taste. By including the ethanol production inhibitor in the process, a very desirable fermentation may be obtained without any ethanol production or at least with limited ethanol production. Therefore, a fermented non-alcoholic beverage may be obtained without removing alcohol e.g. by distillation, which is typically applied for conventional non-alcohol wine. By avoiding e.g. distillation, the flavor composition of the fermented beverage may be kept intact. Some flavor compounds may be removed by the distillation, which can be avoided for the present invention to keep the original flavor composition.

[0010] Also, a further advantage of the invention may be that a non-alcoholic beverage may be obtained with a lower energy consumption, decreased water use, decreased use of raw materials and more cost efficiently since removal of alcohol by e.g. distillation may be superfluous. Thus, the produced beverage may obtain a content of alcohol below 0.5% ABV without any post-fermentation alteration of the alcohol content.

[0011] It should of course be understood that the step of inoculating the starting material involves adding the at least one malolactic bacterial strain.

[0012] Also, in the present context it should be understood that the step of inoculating the starting material with at least one bacterial strain comprises adding the at least one bacterial strain and may further comprise, according to an embodiment, mixing, e.g. to obtain a substantially homogeneously mixed composition.

[0013] According to an embodiment of the invention, the step d starts at the moment where the starting material is inoculated and any further conditions necessary for fermentation are fulfilled, e.g. by adjusting the temperature to a level supporting bacterial growth.

[0014] It is noted that it is possible to reduce the amount of alcohol significantly and even to a degree that the content is below 0.5 % ABV even with a surprisingly low amount of ethanol production inhibitor, such as between 0.1 to 1.0 g / L.

[0015] According to an embodiment of the invention, step b is performed prior to step d.

[0016] According to an embodiment of the invention, step d comprises fermenting the starting material including the at least one malolactic bacterial strain and the at least one ethanol production inhibitor.

[0017] According to an embodiment of the invention, step b comprises adding the at least one ethanol production inhibitor to the starting material having a pH value of at least 3.0 at the time of addition, such as at least 3.5 at the time of addition, such as a pH value of at least 4.0 at the time of addition. According to an embodiment of the invention, step d results in a change of pH value, wherein step b occurs when no more than 50 % of the change of pH value has occurred, such as no more than 20 % of the change of pH value has occurred.

[0018] Thus, according to the above embodiment, step b comprises adding the at least one ethanol production inhibitor to the starting material when it has a pH value of at least 0.5*pH_initial + 0.5*pH_final, such as at least 0.8*pH_initial + 0.2*pH_final, wherein pHJnitial is the pH value of the starting material before step c, and wherein the pH_final is the lowest pH value during step d.

[0019] According to an embodiment of the invention, step d results in a change of brix value, wherein step b occurs when no more than 50% of the change of brix value has occurred, such as no more than 20% of the change of brix value has occurred.

[0020] Thus, according to the above embodiment, step b comprises adding the at least one ethanol production inhibitor to the starting material when it has a pH value of at least 0.5*brix_initial + 0.5*brix_final, such as at least 0.8*brix_initial + 0.2*brix_final, wherein brix_initia I is the brix value of the starting material before step c, and wherein the brix_final is the lowest brix value during step d.

[0021] According to an embodiment of the invention, step b comprises adding the at least one ethanol production inhibitor to the fermentation mixture having an alcohol content of no more than 8.0 % ABV, such as no more than 3.0% ABV, such as no more than 1.0 % ABV, such as no more than 0.5 % ABV.

[0022] According to an embodiment of the invention, the fermentation is performed for a duration of which at least 50% is after step b, such as at least 80% is after step b, such as at least 90% is after step b, such as at least 95% is after step b, such as at least 98% is after step b, such as at least 99% is after step b.

[0023] It is noted that the above embodiment refers to the duration of the fermentation. In an example where the duration of the whole fermentation is 10 days, and at least 50% of the duration of the fermentation is after step b, this means that the step b occurs not earlier than after 5 days, i.e. such that at least 5 days of the fermentation is after step b. According to an embodiment of the invention, step d results in a change in cfu count, wherein step b occurs when no more than 50% of the change in the cfu count has occurred, such as no more than 20% of the change in the cfu count has occurred.

[0024] Thus, according to the above embodiment, step b comprises adding the at least one ethanol production inhibitor to the starting material when it has a cfu value of no more than 0.5*cfu_initial + 0.5*cfu_final, such as no more than 0.8*cfu_initial + 0.2*cfu_final, wherein cfujnitial is the cfu value of the starting material before step c, and wherein the cfu_final is the highest cfu value during step d.

[0025] According to an embodiment of the invention, the process comprises a primary fermentation and optionally a secondary fermentation, and wherein the ethanol production inhibitor is added before initiation of the primary fermentation.

[0026] According to an embodiment of the invention, the grape extract is made from one or more grape varieties selected from the group comprising Chardonnay, Sauvignon Blanc, Pinot Gris, Pinot Noir, Semilion, Gewurztraminer, Chenin Blanc, Pinot Blanc, Viognier, Roussanne, Marsanne, Muscat, Riesling, Silvaner, Muller-Thurgau, Gruner Veltliner, Torrontes, Garganega, Verdicchio, Sangiovese, Tempranillo, Zinfandel, Pinotage, Cabernet Franc, Merlot, Albarino, Airen, and any combination thereof.

[0027] It should be noted that the above-mentioned grape varieties are just a few examples of the many applicable varieties existing in total.

[0028] According to an embodiment of the invention, fermenting the starting material is performed by the malolactic bacteria.

[0029] According to an embodiment of the invention, fermenting the starting material is a primary fermentation.

[0030] In the present context the term "primary fermentation" refers to a fermentation being performed as the first fermentation, optionally followed by a so-called "secondary fermentation". Consequently, within the present context, a primary fermentation does not necessarily include yeast fermentation, in particular alcohol producing yeast fermentation. According to an embodiment of the invention, fermenting the starting material is a primary fermentation performed by the malolactic bacteria.

[0031] According to an embodiment of the invention, no secondary fermentation is performed.

[0032] In an embodiment, the fermentation is a primary fermentation, and where the primary fermentation is followed by a secondary fermentation.

[0033] According to an embodiment of the invention, a fermentation mixture is obtained from the starting material and the at least one bacterial culture.

[0034] The fermentation mixture of the above embodiment refers to the material obtained by mixing at least the starting material and the at least one bacterial culture and throughout the fermentation step.

[0035] According to an embodiment of the invention, the viable yeast count in the fermentation mixture is equal to or below 3.0*10A4 cfu / mL, such as below 2.0*10A4 cfu / mL, such as below 1.56*10 4 cfu / mL, such as below 1.0*10 4 cfu / mL, such as below 3.0*10 3 cfu / mL, such as below 100 cfu / mL, such as below 10 cfu / mL.

[0036] According to an embodiment of the invention, step b is performed when the cfu count of ethanol-producing microorganisms in the starting material is between 0 and 3.0*10A4 cfu / mL, such as between 0 and 2.0*10A4 cfu / mL, such as between 0 and 1.56*10A4 cfu / mL, such as between 0 and 1.0*10 4 cfu / mL, such as between 0 and 3.0*10 3 cfu / mL, such as between 0 and 100 cfu / mL, such as between 0 and 10 cfu / mL.

[0037] According to an embodiment of the invention, the viable yeast count in the fermentation mixture is equal to or below 1.56*10A4 cfu / mL.

[0038] According to an embodiment of the invention, the inoculation is performed once.

[0039] According to an embodiment of the invention, the inoculation is a bacterial inoculation.

[0040] According to an embodiment of the invention, the inoculation is performed once and wherein the inoculation is a bacterial inoculation.

[0041] According to an embodiment of the invention, no yeast is added. According to an embodiment of the invention, no viable yeast is added.

[0042] According to an embodiment of the invention, no viable Saccharomyces yeast is added.

[0043] According to an embodiment of the invention, the at least one bacterial strain comprises a malolactic bacterial strain, such as consists of a malolactic bacterial strain.

[0044] According to an embodiment of the invention, the at least one malolactic bacterial strain includes lactic acid bacteria.

[0045] According to an embodiment of the invention, the at least one bacterial strain is selected from the group consisting of Lactobacillus, Leuconostoc, Pediococcus, Lactococcus, and Streptococcus, Aerococcus, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Tetragenococcus, Vagococcus Weissella, and any combination thereof.

[0046] According to an embodiment of the invention, the at least one malolactic bacterial strain is selected from the group consisting of one or more Lactobacillus plantarum ssp and / or one or more Oenococcus oeni ssp.

[0047] According to an embodiment of the invention, the at least one bacterial strain is Lactobacillus plantarum ssp.

[0048] According to an embodiment of the invention, the at least one bacterial strain is Oenococcus oeni ssp.

[0049] According to an embodiment of the invention, the at least one bacterial strain comprises a non-malolactic bacterial strain, such as a non-malolactic bacterial strain selected from the group consisting of Actinobacillus succinogenes, Anaerobiospirillum succiniciproducens, Escherichia coli, Acetobacteraceae (e.g. acetobacter and gluconobacter), Pseudomonadaceae, Enterobacteriaceae, and any combination thereof

[0050] According to an embodiment of the invention, the at least one malolactic bacterial strain is added in an amount in the range of from 2.0*10 8 to 1.5*10 10 cfu / L starting material.

[0051] It is noted that when using notation such as 2.0E10 or 2.0E+10, this of course refers to 2.0*10 10. This is also often referred to as the so-called E-notation. According to an embodiment of the invention, the starting material has a Brix value in the range from 0.5 to 75.

[0052] According to an embodiment of the invention, the starting material has a Brix value in the range from 2 to 20.

[0053] According to an embodiment of the invention, the starting material has a Brix value in the range from 5 to 10.

[0054] According to an embodiment of the invention, the Brix value of the non-alcoholic beverage is in the range from 0.25 to 30.

[0055] According to an embodiment of the invention, the Brix value of the non-alcoholic beverage is in the range from 2 to 7.

[0056] According to an embodiment of the invention, the process further comprises a step of adding nutrients to the fermentation mixture.

[0057] According to an embodiment of the invention, the process further comprises a step of adding stability enhancers to the fermentation mixture.

[0058] According to an embodiment of the invention, the process further comprises a step of adding regulators of osmotic pressure to the bacteria before inoculation.

[0059] According to an embodiment of the invention, fermenting the starting material comprises conversion of malic acid into lactic acid and carbon dioxide.

[0060] According to an embodiment of the invention, fermenting the starting material does not produce alcohol.

[0061] In the present context a fermentation may be understood as a sensorial change due to microorganisms (organoleptic changes).

[0062] According to an embodiment of the invention, fermenting the starting material is anaerobic. In the present context, anaerobic means that the process may be totally anaerobic but also essentially anaerobic in the sense that the containers in which the fermentation is performed may be closed but that a very little amount of oxygen may be present in the starting material from the start and during the fermentation or a part of the fermentation. It may sometimes be difficult in practice to ensure that the fermentation mixture is totally free of exposure to oxygen.

[0063] According to an embodiment of the invention, the fermentation is performed for a duration which is no more than 15 days.

[0064] According to an embodiment of the invention, the fermentation is performed for a duration which is no more than 45 days.

[0065] According to an embodiment of the invention, fermenting the starting material is performed for a duration of 1 hour to 45 days, such as 2 hours to 20 days, such as 3 hours to 15 days, such as 4 hours to 10 days.

[0066] According to an embodiment of the invention, the ethanol production inhibitor comprises one or more selected from the group consisting of sorbic acid, sorbic acid salts, benzoic acid, benzoic acid salts, and any combination thereof.

[0067] According to an embodiment of the invention, the ethanol production inhibitor comprises one or more selected from the group consisting of sorbic acid, potassium sorbate, benzoic acid, sodium benzoate, potassium benzoate, calcium benzoate, ethyl- p-hydroxybenzoate, sodium ethyl p-hydroxybenzoate, methyl p-hydroxybenzoate, sodium methyl p-hydroxybenzoate, and any combination thereof.

[0068] According to an embodiment of the invention, the ethanol production inhibitor comprises potassium sorbate and / or sodium benzoate.

[0069] According to an embodiment of the invention, the ethanol production inhibitor is a yeast fermentation inhibitor.

[0070] According to an embodiment of the invention, the ethanol production inhibitor is added in an amount of between 0.01 and 5 g / L, such as between 0.01 and 1 g / L. It is noted that it is possible to reduce the amount of alcohol significantly and even to a degree that the content is below 0.5 % ABV even with a surprisingly low amount of ethanol production inhibitor, such as between 0.1 to 1.0 g / L.

[0071] According to an embodiment of the invention, the ethanol production inhibitor is added in an amount of between 0.1 and 0.5 g / L.

[0072] According to an embodiment of the invention, the process further comprises adding fermentation aids and / or flavouring ingredients.

[0073] According to an embodiment of the invention, the flavouring ingredients include oak chips.

[0074] According to an embodiment of the invention, fermentation aids and / or flavouring agents are added before or during fermentation.

[0075] According to an embodiment of the invention, the flavouring ingredients include tannins.

[0076] According to an embodiment of the invention, the flavouring ingredients comprise botanical flavour ingredients and / or natural aromas.

[0077] According to an embodiment of the invention, the botanical flavour ingredients and / or natural aromas are derived from fruit, berries, herbs, spices, aromatic plant parts, or any combination thereof.

[0078] According to an embodiment of the invention, the process further comprises a step of fining the fermentation mixture after fermentation.

[0079] According to an embodiment of the invention, the step of fining the fermentation mixture includes the addition of fining agents such as organic fining components or such as mineral fining components.

[0080] According to an embodiment of the invention, the fining agents comprise Kieselsol and / or Chitosan.

[0081] According to an embodiment of the invention, the organic fining components comprise egg whites, casein, gelatine, chitin, isinglass, and any combination thereof. According to an embodiment of the invention, the mineral fining components comprise bentonite, charcoal, potassium ferrocyanide, silica, kaolin, and any combination thereof.

[0082] According to an embodiment of the invention, the process further comprises a step of sterilizing the fermentation mixture after fermentation.

[0083] According to an embodiment of the invention, the fermentation mixture comprises sterile filtration and / or pasteurization.

[0084] According to an embodiment of the invention, the process further comprises adding sulfites to the fermentation mixture after fermentation.

[0085] According to an embodiment of the invention, the process further comprises bottling, canning, boxing or kegging the non-alcoholic beverage.

[0086] According to an embodiment of the invention, the process further comprises a step of filtering the fermentation mixture after fermentation.

[0087] According to an embodiment of the invention, the fermentation mixture after fermentation comprises a first filtration step and a second filtration step.

[0088] According to an embodiment of the invention, the first filtration step is performed using a filter having a pore size of at least 2 micron.

[0089] According to an embodiment of the invention, the second filtration step is performed using a filter having a pore size between 0.1 and 2.0 micrometers such as between 0.2 and 1.5 micrometers, such as between 0.3 and 1.0 micrometers, such as between 0.1 and 0.5 micrometers, such as between 0.2 and 0.5 micrometers, such as between 0.3 and 0.5 micrometers, such as between 0.4 and 0.6 micrometres.

[0090] According to an embodiment of the invention, the starting material has been subject to enzymatic treatment prior to inoculating the starting material with at least one bacterial strain.

[0091] According to an embodiment of the invention, the starting material is fermented until the pH is in the range of from 2 to 4. According to an embodiment of the invention, the process further comprises a step of adding nutrients and / or growth support agents to the fermentation mixture.

[0092] According to an embodiment of the invention, the nutrients and / or growth support agents comprise inactivated yeast, protein hydrolysate, amino acids, polysaccharides, bacterial cell wall polypeptides, parietal polysaccharides, cofactors, vitamins, and any combination thereof.

[0093] According to an embodiment of the invention, the nutrients and / or growth support agents comprise Enartis Nutriferm ML and / or Enartis Nutriferm Osmobacti.

[0094] According to an embodiment of the invention, the non-alcoholic beverage is a nonalcoholic wine analogue.

[0095] The invention further relates to a process for producing a non-alcoholic beverage comprising the steps of a. Providing a starting material consisting of a grape extract; wherein the starting material has a Brix value in the range from 5 to 10, such as from 6 to 8; b. Adding at least one malolactic bacterial strain to the starting material; wherein said bacterial strain is selected from the group consisting of one or more Lactobacillus plantarum ssp and / or one or more Oenococcus oeni ssp; c. Fermenting the starting material comprising the at least one malolactic bacterial strain until the pH is in the range of from 2-4; d. Adding Potassium sorbate in the range from 0.1 to 1 g / L; e. Adding flavour ingredients comprising adding oak chips in an amount in the range from 0.01 to 2 g / L; f. Fining the non-alcoholic beverage; g. Filtering the non-alcoholic beverage; h. Bottling the non-alcoholic beverage; thereby achieving a non-alcoholic beverage having an alcohol content of not more than 0.5% ABV.

[0096] The invention further relates to a process for producing a non-alcoholic beverage comprising the steps of a. Providing a starting material comprising grape extract; wherein the starting material has a Brix value in the range from 6 to 10; b. Adding at least one malolactic bacteria to the starting material; c. Fermenting the starting material comprising the at least one malolactic acid bacteria until the pH is in the range of from 2 to 3.5; d. Addition of a least one agent which prevents yeast fermentation, for example a sequestrant, to the fermentation; e. Optionally adding at least one flavour ingredient, thereby achieving a non-alcoholic beverage having an alcohol content of not more than 0.5% ABV.

[0097] According to an embodiment of the invention, the ethanol production inhibitor comprises potassium sorbate in the range from 0.1 to 1 g / L.

[0098] According to a further embodiment of the invention, the ethanol production inhibitor comprises sodium benzoate in the range from 0.1 to 1 g / L.

[0099] According to an embodiment of the invention, the process further comprises at least one step selected from the group consisting of

[0100] - Fining the non-alcoholic beverage;

[0101] - Filtering the non-alcoholic beverage;

[0102] - Bottling the non-alcoholic beverage; and any combination thereof.

[0103] The invention further relates to use of the malolactic bacterial strain in a process according to the invention or any of its embodiments.

[0104] The invention further relates to a non-alcoholic beverage obtainable or obtained by the process according to the invention or any of its embodiments.

[0105] The invention further relates to a non-alcoholic beverage having an alcohol content of not more than 0.5% ABV alcohol, a pH in the range of 2-4 and a Brix value of 5 to 10 °Brix.

[0106] The invention further relates to a non-alcoholic beverage having an alcohol content of not more than 0.5% ABV alcohol, the beverage comprising fermented grape extract, wherein the fermented grape extract is obtained without yeast fermentation. In an embodiment of the invention, the non-alcoholic beverage of the invention described above is obtainable by the process according to the invention or any of its embodiment.

[0107] The invention further relates to a non-alcoholic beverage having an alcohol content of not more than 0.5% ABV alcohol, the beverage comprising fermented grape extract, wherein the beverage is made without removing alcohol.

[0108] In an embodiment of the invention, the non-alcoholic beverage of the invention described above is obtainable by the process according to the invention or any of its embodiment.

[0109] The invention further relates to a non-alcoholic beverage having an alcohol content of not more than 0.5% ABV alcohol, the beverage comprising fermented grape extract and an ethanol production inhibitor of less than 0.5 g / L.

[0110] In an embodiment of the invention, the non-alcoholic beverage of the invention described above is obtainable by the process according to the invention or any of its embodiment.

[0111] The inventors have identified a process of producing a non-alcoholic beverage, using malolactic bacteria. The process provides an efficient method of producing beverages with taste profiles similar to wine while avoiding the alcohol produced by yeast fermentation. The method is more sustainable from a climate and energy point of view as it provides a more efficient use of resources. The invention thus relates to a process of producing a non-alcoholic beverage, to the non-alcoholic beverage obtained or obtainable by the process, and to the use of malolactic bacteria in said process.

[0112] A further aspect the invention relates to a process for producing a non-alcoholic beverage comprising the steps of a. Providing a starting material comprising grape juice; wherein the starting material has a Brix value in the range from 6 to 10; b. Adding at least one malolactic bacteria to the starting material; c. Fermenting the starting material comprising the at least one malolactic bacteria until the pH is in the range of from 2 to 4; d. Addition of a least one agent which prevents yeast fermentation, for example a sequestrant, in fermentation mixture; e. Optionally adding at least one flavour ingredient, thereby achieving a non-alcoholic beverage having an alcohol content of less than 0.5% ABV.

[0113] A yet a further aspect, the invention relates to the use of malolactic bacteria in a process according to an embodiment of the invention.

[0114] In yet a further aspect, the invention relates to a non-alcoholic beverage obtainable or obtained by the process. The non-alcoholic beverages, according to an embodiment of the invention, may be referred to as wine analogues. As used herein, the term wine analogue refers to a beverage based on fermented grape juice where only low amounts of alcohol are produced. These wine analogues resemble wine in look and taste but have an alcohol content of not more than 0.5 % ABV.

[0115] BRIEF DESCRIPTION OF THE FIGURES

[0116] Figure 1 shows the pH developments throughout fermentation for 1 sample production run (See Example 6, Results). Initial pH is 4-4.15 and throughout the 10 days of fermentation it drops to within the range of 3.1-3.2. Further experiments showed an initial steep drop in pH, roughly 1 pH level over the first 4 days, followed by a flattening out of the pH decrease, with about 0.3 pH level over the following 2-3 days.

[0117] Figure 2 shows the changes in °Brix throughout fermentation for a given production of 7°Brix base substrate. Figure 2 shows very little to no variation in °Brix throughout fermentation, maximum difference recorded between initiation and day 10 has been 0.2 °Brix. Later experiments showed a higher decrease in Brix value of up to about

[0118] 1 °Brix.

[0119] Figure 3 shows the changes in organoleptic properties throughout fermentation. Displayed from 0-5 Arbitrary units., (0 being lowest amounts; and 5 being highest amount). Solid line- Sweetness; Dashed line- Dryness; Dotted line- Sourness. The profile shown with increased sourness and dryness and decreased sweetness reflects a desirable organoleptic profile.

[0120] Figure 4 shows a process for producing non-alcoholic beverage.

[0121] DETAILED DESCRIPTION OF THE INVENTION

[0122] Definitions

[0123] The term alcohol and ethanol are used interchangeably herein and refer both to ethanol unless otherwise specified. The percentage alcohol is alcohol by volume, unless specifically indicated otherwise.

[0124] The term alcohol by volume (ABV) as used herein refers to a standard measure of how much alcohol (i.e. ethanol) is contained in a given volume of an alcoholic beverage (expressed as a volume percent). It is defined as the number of millilitres (mL) of pure ethanol present in 100 mL of solution at 20°C. The use of the terms "a" and "an" and "the" and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms "comprising", "comprises", "having”, "including", "at least" and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to,") unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method for referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any nonclaimed element as essential to the practice of the invention.

[0125] Reference to "about" a value or parameter herein includes embodiments that are directed to that value or parameter per se. For example, description referring to "about X" includes the embodiment "X". When used in combination with measured values, "about" includes a range that encompasses at least the uncertainty associated with the method of measuring the particular value and can include a range of plus or minus two standard deviations around the stated value.

[0126] As used herein and in the appended claims, the singular forms "a," "or," and "the" include plural referents unless the context clearly dictates otherwise.

[0127] It is understood that the embodiments described herein include "consisting" and / or "consisting essentially of" embodiments. As used herein, except where the context requires otherwise due to express language or necessary implication, the word "comprise" or variations such as "comprises" or "comprising" is used in an inclusive sense, i.e. to specify the presence of the stated features but not to preclude the presence or addition of further features in various embodiments.

[0128] DESCRIPTION

[0129] The present invention relates to a process of producing a non-alcoholic beverage. The non-alcoholic beverage according to an embodiment of the invention is a grape-based beverage, produced by fermentation and having less than 0.5 % alcohol by volume (ABV).

[0130] In some methods of producing non-alcoholic wine, a traditional wine comprising ethanol is produced and the ethanol later removed, a process known as dealcoholization. This process is time- and resource demanding and may produce undesirable side-products in the wine. The process of the invention is efficient and provides an alternative method of producing a non-alcoholic beverage which avoids at least these two drawbacks of the known methods.

[0131] Some embodiments of the invention relate to a non-alcoholic beverage which is a wine analogue.

[0132] Starting material

[0133] The process of the invention comprises a first step of providing a starting material comprising or consisting of grape extract. Grape extract should, in the present context, be understood as grape derivatives in any suitable form, such as one or more of the following: Grape concentrate, grape must, grape juice, grape press, grape powder and / or dilutions of the before-mentioned.

[0134] The product of the process of the invention is a non-alcoholic beverage, such as a nonalcoholic wine analogue. Therefore, the starting material may be any starting material useful in producing an alcoholic wine.

[0135] The grape extract suitable as a starting material in the process of the invention may comprise at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% or at least 95% extract from grapes. In one embodiment, the grape extract comprises at least 90% grape, for example the grape extract may comprise at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or for example may consist of extract from grapes.

[0136] In particular embodiments of the invention, the starting material is a grape concentrate. Grape concentrates suitable for producing wine analogues according to an embodiment of the invention may be concentrates from one or more grape varieties selected from the group consisting of: Chardonnay, Sauvignon Blanc, Pinot Gris, Pinot Noir, Semilion, Gewurztraminer, Chenin Blanc, Pinot Blanc, Viognier, Roussanne, Marsanne, Muscat, Riesling, Silvaner, Muller-Thurgau, Gruner Veltliner, Torrontes, Garganega, Verdicchio, Sangiovese, Tempranillo, Zinfandel, Pinotage, Cabernet Franc, Merlot, Albarino or Airen.

[0137] Some embodiments relate to situations where the starting material comprises extract from table grapes, as well as specific grape varieties, such as one or more mentioned above. Other embodiments relate to situations where the starting material does not comprise extract from table grapes.

[0138] Particular embodiments relate to situations where the starting material is made by diluting a grape concentrate with water to achieve the suitable °Brix. Thus, the starting material may comprise or consist of grape concentrate diluted with water.

[0139] In some embodiments, the starting material is sterile.

[0140] In accordance with the definitions above, fruit extract should be understood as derivatives of any fruit suitable for making beverages according to an embodiment of the invention, such as one or more of the following: Fruit concentrate, fruit must, fruit juice, fruit press and / or dilutions of the before-mentioned. Fruit suitable for making beverages according to an embodiment of the invention may for instance be grapes, apple, pear, plum, cherries, berries and any combinations of these.

[0141] Brix

[0142] The starting material in the process according to an embodiment of the invention has a Brix value in the range of from 6 to 10 °Brix. This may be achieved by diluting a grape must or grape juice with water until the selected Brix value is achieved.

[0143] Brix value is defined as grams of soluble solids per 100 g of solution and is expressed as degrees Brix (°Brix). In fruit juices, e.g. grape juice, the most abundant solid is sugar (sucrose and fructose), and the Brix value of a sample thus approximates the sugar content of that sample.

[0144] The inventors have surprisingly discovered that the Brix value of the starting material is important for the development of flavour.

[0145] If Brix values are too high or too low in the starting material, the flavour development will not be optimal (see Example 1). In an embodiment, a good flavour development can be achieved when the starting material has a Brix value in the range of from 6-10 °Brix.

[0146] In some embodiments of the invention, the starting material has a Brix value in the range of 5-15, 6-10, 7-10, 8-10, 9-10, or for example 5-10, 8-15, 6-9, 6-8 or 6-7 °Brix. In particular embodiments, the starting material has a Brix value in the range from 6 to 10, such as from 6 to 8 °Brix.

[0147] Malolactic bacteria The process of the invention comprises a step of adding at least one malolactic bacterial strain to the starting material, in order to perform a malolactic fermentation. The fermentation process contributes to development of the flavour of the non-alcoholic beverage while avoiding ethanol production.

[0148] In the present context, the term 'malolactic bacterial strain' is defined as a group of similar malolactic bacterial cells which produce lactic acid as their major metabolic end product. Thus, in the present context, malolactic bacterial strains include lactic acid bacterial strains, and malolactic bacteria include lactic acid bacteria. In other words, within the context of this patent application, lactic acid bacteria are considered a subgroup of malolactic bacteria, and malolactic fermentation covers fermentation performed by both malolactic and lactic acid bacteria.

[0149] The malolactic fermentation of the invention contributes to the flavour profile, while not producing ethanol. In conventional winemaking, malolactic fermentation may occur either during or after the primary, alcohol-producing yeast fermentation. In contrast, in the present invention, the malolactic fermentation is the primary, or only, fermentation.

[0150] The malolactic bacteria suitable for use in the invention may be, for example, one or more bacteria of the genera Lactobacillus, Leuconostoc, Pediococcus, Lactococcus, and Streptococcus, Aerococcus, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Tetragenococcus, Vagococcus, and Weissella.

[0151] Particular embodiments of the invention relate to where the at least one malolactic bacterial strain comprises or consists of one or more of the Oenococcus oeni and / or Lactobacillus plantarum strains such as for example consists of a Lactobacillus plantarum strain.

[0152] In further embodiments, the at least one malolactic bacterial strain is selected from the group consisting of Chr. Hansen VINIFLORA® CH16 {Oenococcus oeni ssp), Chr. Hansen VINIFLORA® CH35 {Oenococcus oeni ssp , Chr. Hansen VINIFLORA® Oenos 2.0 {Oenococcus oeni ssp), Chr. Hansen VINIFLORA® Nova Plus {Lactobacillus plantarum ssp), or combinations thereof. In further embodiments, the malolactic bacterial strain comprises Chr. Hansen VINIFLORA® Nova Plus {Lactobacillus plantarum ssp). In even further embodiments, the malolactic bacteria added to the starting material consists of Chr. Hansen VINIFLORA® Nova Plus {Lactobacillus plantarum).

[0153] In further embodiments, the at least one malolactic bacterial strain comprises or consists of the DSM L. plantarum DSM 33452 strain.

[0154] The invention relates, in particular embodiments, to where malolactic bacteria is added in an amount in the range of from 2.0E+8 to 1.5E+10 cfu / L, for example in the range from 2.0E+8 to 1.0e+10, 2.0E+8 to 1.0E+9, or 2.0E+8 to 9.0E+8 cfu / L, or 2.0E+8 to 8.0E+8 cfu / L, or for example 4.0E+8 cfu / L.

[0155] The invention relates in some embodiments to where the malolactic bacteria is added in an amount in the range of from 0,001 - 0.075 g malolactic bacteria per litre starting material, where the concentration of malolactic bacteria is 2.0E+11 cfu / g.

[0156] Fermentation

[0157] The process of the invention comprises malolactic fermentation of a starting material. In particular embodiments, the fermentation is performed at an ambient temperature of not more than 25°C. In some embodiments, the fermentation is performed at an ambient temperature in the range from 10°C-25°C, 12°C-25°C, 15°C-25°C, 18°C- 25°C, 20°C-25°C, 15°C-23°C, or 18°C-23°C. In a preferred embodiment the fermentation is performed at an ambient temperature in the range of 18°C to 23°C; or, for example, at 18°C, 19°C, 20°C, 21°C, 22°C, or 23°C. In an additional or alternative embodiment, the fermentation is performed at an ambient temperature of 22°C to 23°C. The inventors have identified that higher temperatures, such as temperatures of above 25°C may lead to development of unfavourable flavours, such as acetone.

[0158] In an alternative embodiment, fermentation is performed by non-saccharomyces yeast. Non-saccharomyces yeast should be understood as yeast which do not produce significant amounts of alcohol and do not belong to the Saccharomyces genus, in particular the Saccharomyces cerevisiae (S. cerevisiae) species.

[0159] Bacterial nutrition and growth support

[0160] The process of the invention may further comprise a step of adding nutrients to the fermentation mixture. They nutrients may be any composition suitable for supporting growth of the malolactic bacteria.

[0161] The nutrients may, for example, comprise one or more of inactivated yeast, protein hydrolysate, amino acids, polysaccharides, bacterial cell wall polypeptides, parietal polysaccharides, cofactors and vitamins, or combinations thereof.

[0162] In this context, inactivated yeast should be understood as yeast cells which are not viable, living, alive, functioning or active or yeast which have been defunctionalized, killed, or autolyzed. The term also includes inactivated yeast derivatives. In essence, inactivated yeast cells do not produce alcohol but may prevent oxidation, reduce the use of sulphites, enhance sensory properties, stabilize colour and / or improve the balance, softness and freshness of the beverage. Accordingly, active or viable yeast should be understood as yeast cells which are living, alive, functioning and active. Viable yeast may or may not produce alcohol depending on the specific species.

[0163] Other growth support agents include, for example, agents which adsorb toxic compounds from fermentation, such as e.g. cellulose.

[0164] In particular embodiments, nutrients and / or growth support agents are provided by addition of Enartis Nutriferm ML and / or Enartis Nutriferm Osmobacti.

[0165] These agents increase the availability of nutrients in the fermentation mixture, support bacterial cell growth, and adsorb compounds that are toxic to the bacteria. The combined use helps to promote the dominance of the inoculated strain and reduces the duration of the malolactic fermentation.

[0166] In particular embodiments, the step of adding nutrients is performed at initiation of fermentation.

[0167] In particular embodiments, the process according to an embodiment of the invention is performed using reverse osmosis water.

[0168] PH

[0169] As the fermentation proceeds, the pH progressively decreases. The inventors have discovered that an advantageous taste development occurs when the fermentation is allowed to proceed such that the fermentation liquid reaches a pH in the range from pH 2 to pH 4, such as in the range from 2 to 3.5, for example from 2.5 to 3.5. Particular embodiments relate to where the pH is in the range from 2.7 to 3.5, 2.8 to 3.4, or for example in the range of from 2.9 to 3.2.

[0170] Without wishing to be bound by theory, it is thought that the pH reflects the optimal state of the fermentation. It reflects the balance between allowing beneficial conversion processes to take place for as long as possible, but terminating before bacterial lysis leads to degradatory effects on the fermentation mixture, such as development of off- flavours (see Example 5).

[0171] Fermentation time

[0172] The fermentation step in the process of the invention may be performed for a predetermined time interval. Performing the fermentation step for a predetermined time interval improves simplicity and allows for simpler automation of the fermentation process.

[0173] Accordingly, in some embodiments of the invention, the fermentation step is performed for no more than 15 days - for example, in the range from 5 to 15 days, such as 5 to 14, 5 to 13, 5 to 12, 5 to 11, or for example 6 to 16, 6 to 14, 6 to 12; such as for example in the range from 8 to 11 days. Particular embodiments of the invention relate to where fermentation is no more than 10 days.

[0174] Alcohol percentage

[0175] The process according to an embodiment of the invention yields a non-alcoholic beverage having an alcohol content of not more than 0.5 % ABV.

[0176] The labelling of beverages as non-alcoholic varies according to jurisdiction. In the context of the present invention, beverages having an alcohol content of not more than 0.5 % ABV are considered non-alcoholic.

[0177] The process according to an embodiment of the invention is particularly good at making very low levels of alcohol. Thus, particular embodiments of the invention relate to where the non-alcoholic beverage has an alcohol content of not more than 0.5% ABV, such as not more than 0.4%, not more 0.3%, not more than 0.2%, not more than 0.1% ABV, or for example 0.08% or less ABV. In other embodiments, the non-alcoholic beverage has an alcohol content of essentially 0% ABV. Further embodiments relate to where the non-alcoholic beverage, according to an embodiment of the invention, has an alcohol content in the range from 0.01 to 0.5%, such as 0.02 to 0.4%, such as 0.03 to 0.3%, or 0.01 to 0.08% ABV.

[0178] Prevention of alcohol production

[0179] No alcohol is produced by malolactic fermentation. However, in order to prevent alcohol-producing side fermentations, for example from yeast contaminations, the process of the invention comprises a step of adding at least one agent which inhibits the growth of alcohol-producing yeast within the fermentation mixture. Accordingly, such agent is an alcohol production inhibitor.

[0180] Inhibiting the growth of alcohol-producing yeast may be achieved in any suitable manner compatible with food products. For example, agents that will bind to alcohol- producing yeast cells and sequester them or in any other manner will prevent the alcohol-producing yeast from growing and / or producing alcohol are considered alcohol production inhibitors. One example of an alcohol production inhibitor is potassium sorbate.

[0181] In particular, addition of the at least one alcohol production inhibitor prevents alcohol production driven by contaminating microorganisms such as alcohol-producing yeast. Potassium sorbate

[0182] Particular embodiments of the invention relate to where the alcohol production inhibitor comprises or consists of potassium sorbate and / or sodium benzoate.

[0183] In yet further embodiments, the alcohol production inhibitor consists of potassium sorbate. In a particular embodiment of the invention, the step of adding the alcohol production inhibitor is performed early in the process. In some embodiments, the alcohol production inhibitor is added substantially at the same time as inoculation with malolactic bacteria. In additional or alternative embodiments, the alcohol production inhibitor is added about 1 hr, 2 hrs, 3 hrs, 4 hrs, 5 hrs or 6 hrs after inoculation with malolactic bacteria. In an additional or alternative embodiment, the alcohol production inhibitor is added after less than 1 hr after inoculation with malolactic bacteria, for example after 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min or 55 min.

[0184] In particular embodiments, potassium sorbate is added in an amount in the range of from 0.1 to 1 g / L, such as in the range from 0.1 to 0.75 g / L, 0.1 to 0.6 g / L, 0.1 to 0.5 g / L, 0.1 to 0.3 g / L. Particular embodiments relate to a method according to an embodiment of the invention wherein the amount of potassium sorbate is in the range of from 0.2 to 0.5 g / L, such as, for example, about 0.25 g / L.

[0185] Flavourings

[0186] The process according to an embodiment of the invention optionally comprises at least one step of adding at least one flavouring ingredient.

[0187] The term flavour ingredient as used herein refers to any ingredient which by its addition imparts a flavour to the non-alcoholic beverage produced by the method.

[0188] Some embodiments of the invention relate to where flavouring ingredients are added. Such ingredients may be any edible flavour ingredients. The selection of flavour ingredients is within the skill of a person with experience in the field of fermented beverages and wine.

[0189] In some embodiments, the flavouring ingredient imparts an oak flavour. In some embodiments the flavouring ingredient may comprise or consist of for example tannins and / or oak chips. For example, oak chips may be added in the fermentation process.

[0190] In some embodiments, the flavouring ingredients may comprise or consist of for example a botanical flavour ingredient and / or natural aromas. Botanical flavour ingredients and / or natural aromas as used herein refers to flavour ingredients derived from fruit, berries, herbs, spices, any other aromatic plant part, or any combination thereof.

[0191] In particular embodiments, flavouring ingredients which impart an oak flavour is combined with one or more additional flavouring ingredients, which may comprise or consist of one or more botanical flavouring ingredients and / or natural aromas.

[0192] Regardless of any further flavouring, the process of the invention may be described as providing an optimal non-alcoholic wine-analogue base, to which further and / or alternative flavouring ingredients may be added such as natural aromas. Sweetness

[0193] The process, according to an embodiment of the invention, optionally comprises a step of managing sweetness. Thus, in additional or alternative embodiments, the one or more flavouring ingredients may comprise one or more of sugar, non-caloric sweeteners, sweetness modulators, or sweetener enhancers.

[0194] In one embodiment, the flavouring ingredient is a sugar, preferably sucrose.

[0195] While adding sugar is one alternative embodiment of the present invention, preferred embodiments of the invention relate to where no additional sucrose is added as flavouring ingredient.

[0196] Fining

[0197] The process according to an embodiment of the invention optionally comprises at least one step of adding at least one fining agent.

[0198] Fining agents, as used herein, refers to any substance or composition which may be added to the beverage to create an adsorbent, enzymatic, or ionic bond with particles suspended in the beverage such that these precipitate more readily. Organic components which may be used as fining agents include egg whites, casein, gelatine, chitin and isinglass. Solid and / or mineral components, which may be used as fining agents include bentonite, charcoal, potassium ferrocyanide, silica, kaolin, and combinations thereof.

[0199] Particular embodiments relate to where fining is done by addition of Kieselsol and / or Chitosan. Kieselsol is also known as silica sol and is made from colloidal silica. It primarily works on negatively charged particles. Chitosan is derived from chitin, which is found in the exoskeleton of crustaceans. Chitosan acts on positively charged particles. Particular embodiments relate to the process according to an embodiment of the invention where fining is achieved by treating first with Kieselsol and thereafter with Chitosan. For example, Kieselsol may be added first and Chitosan added after at least one hour followed by anand incubation for at least 24 hours before proceeding to the step of filtration.

[0200] Filtration and bottling

[0201] The process, according to an embodiment of the invention, optionally comprises at least one step of filtering the liquid after fermentation.

[0202] Accordingly, in some embodiments of the invention, the step of the filtration comprises a first filtration and a second filtration.

[0203] In some embodiments, a first filtration is performed to remove larger structures such as cell debris and debris from fermentation. This filtration may be done with any suitable filter, for example having a size of 2 micron or more. In particular embodiments, the first filtration is followed by a second filtration, which may be a filtration of 0.1 to 2, 0.2-1.5, 0.3-1, 0.1-0.5, 0.2-0.5, 0.3-0.5, or 0.4 to 0.6 micrometres. In a preferred embodiment, the second filtration may be a filtration of 0.5 micrometres, resulting in a sterile non-alcoholic beverage.

[0204] The process, according to an embodiment of the invention, optionally further comprises a step of bottling the non-alcoholic beverage.

[0205] Specific embodiments

[0206] Thus, the invention in some embodiments relates to a process for producing a nonalcoholic beverage comprising the steps of a. Providing a starting material consisting of a grape extract; wherein the starting material has a Brix value in the range from 6 to 8, such as a Brix value of 6, such as a Brix value of 7, such as a Brix value of 8; b. Adding at least one malolactic bacterial strain to the starting material; wherein said bacteria is selected from the group consisting of one or more Lactobacillus plantarum subspecies and / or one or more Oenococcus oeni subspecies; preferably in an amount in the range from 2.0E+8 to 8.0E+8 cfu / l, c. Fermenting the starting material comprising the at least one malolactic bacterial strain until the pH is in the range of from 2-4, such as in the range of 2.5-3.5, such as in the range of 3.0-3.5, such as in the range of 2.0-2.5, such as in the range of 3.5- 4.0; d. Addition of Potassium sorbate in the range from 0.01 to 5 g / L, such as in the range from 0.01 to 0.1 g / L, 0.1 to 0.75 g / L, 0.1 to 0.6 g / L, 0.1 to 0.5 g / L, 0.1 to 0.3 g / L or 0.1 to 5 g / L; e. Addition of flavour ingredients, comprising adding oak chips in an amount in the range from 0.01 to 2 g / L, such as in the range from 0.02-1.5 g / L, such as in the range from 0.05 to 1.3 g / L, such as in the range from 0.1 to 1.0 g / L, such as in the range from 0.4 to 0.9 g / L; f. Fining the non-alcoholic beverage; g. Filtering the non-alcoholic beverage; h. Bottling the non-alcoholic beverage; thereby achieving a non-alcoholic beverage having an alcohol content of not more than 0.5 % ABV.

[0207] The invention relates in further particular embodiments to a process for producing a non-alcoholic beverage, which is a wine analogue, the process comprising the steps of a. Providing a starting material consisting of a grape extract; wherein the starting material has a Brix value in the range from 6 to 8; b. Adding at least one malolactic bacterial strain to the starting material; wherein said bacteria comprises or consists of Chr Hansen Viniflora Nova Plus, preferably in an amount in the range from 2.0E+8 to 8.0E+8 cfu / l; c. Fermenting the starting material comprising the at least one malolactic bacterial strain until the pH is in the range of from 2.5-3.5; d. Addition of Potassium sorbate in the range from 0.1 to 1 g / l; e. Addition of flavour ingredients, comprising adding oak chips in an amount in the range from 0.01 to 2 g / I; f. Fining the non-alcoholic beverage; g. Filtering the non-alcoholic beverage; h. Bottling the non-alcoholic beverage; thereby achieving a non-alcoholic beverage having an alcohol content of not more than 0.5%; for example less than 0.08% ABV.

[0208] Use of malolactic bacteria in the process of the invention

[0209] The invention relates in a second aspect to the use of one or more malolactic bacterial strains in the process of the invention. The one or more malolactic bacterial strains for use are described herein above, and the discussion of parameters relating to the process are relevant also to the aspect of use.

[0210] A non-alcoholic beverage

[0211] The invention relates, in a further aspect, to a beverage obtainable or obtained by the method.

[0212] In particular embodiments, the non-alcoholic beverage, according to an embodiment of the invention, is a wine analogue, such as a white wine analogue, a red wine analogue, or a rose wine analogue.

[0213] In further particular embodiments, the beverage according to an embodiment of the invention has a Brix value in the range of from 5 to 10, such as for example 5.3 to 9.8, 6.2 to 8, 6.3. to 7.8, 6.5 to 7.8, 6.7 to 7.8 or for example 6.7 to 7.6, or for example 6.8 to 7.5 °Brix.

[0214] Thus, the invention provides in one embodiment a non-alcoholic beverage according to an embodiment of the invention having an alcohol content of not more than 0.5 % ABV alcohol, a pH in the range of 2-3, an amount of potassium sorbate in the range of from 0.1 to Ig / L and a Brix value of 6 to 8 °Brix.

[0215] Referring to figure 4, a process for producing a non-alcoholic beverage is described according to an embodiment of the invention. First, a grape extract is provided as a starting material SM, e.g. as a grape juice. The grape extract is then subjected to a primary fermentation PF, which is initiated by adding at least one malolactic bacterial strain to the grape extract and thereby inoculating it. Typically, the conditions for the desired fermentation may also be controlled, in particular the temperature of the inoculated grape extract may be carefully controlled. Also, the conditions of the inoculated grape extract may be controlled to be anaerobic or near-anaerobic, e.g. by sealing of the inoculated grape extract in a container to avoid any contact with air or at least minimize any such contact.

[0216] It is noted that the inoculated grape extract may also be referred to as the fermentation mixture.

[0217] It is noted that the inoculated grape extract may in some embodiments also be subjected to mixing, e.g. at inoculation and / or at predefined time intervals throughout the fermentation. In some embodiments, the inoculated grape extract is mixed continuously throughout the fermentation.

[0218] Also, at least one ethanol production inhibitor is added to the grape extract. In some embodiments, the at least one ethanol production inhibitor is added prior to inoculation of the grape extract. In some embodiments, the at least one ethanol production inhibitor is added during inoculation of the grape extract. In some embodiment, the at least one ethanol production inhibitor is added after inoculation of the grape extract. Thus, in the latter case, the at least one ethanol production inhibitor may be added during the fermentation of the grape extract.

[0219] As shown in figure 4, the grape extract fermented in the primary fermentation PF may optionally be subjected to a secondary fermentation SF. In some embodiments the secondary fermentation SF comprises inoculation with a malolactic bacterial strain. In some embodiments, the secondary fermentation SF comprises inoculation with a non- malolactic bacterial strain. In some embodiments the secondary fermentation SF comprises inoculation with a non-Saccharomyces yeast. The aforementioned embodiments may also be combined in order to obtain a combined effect of combined bacterial strains and / or yeast.

[0220] As shown in figure 4, the grape extract fermented in the primary fermentation PF and optionally further fermented in the secondary fermentation SF may be subjected to one or more additional processes FP, such as e.g. racking, fining, filtering etc. Finally, a non-alcoholic beverage having an alcohol content of not more than 0.5% Alcohol By Volume (ABV) is obtained.

[0221] EXAMPLES

[0222] Example 1: Starting concentration

[0223] Materials

[0224] 1. Sugar Concentrations: a. Juice solutions with multiple variants of sugar concentrations: 5 °Brix, 10 °Brix, 15 °Brix, and 20 °Brix. These solutions were prepared by dissolving 65 °Brix grape concentrate (Brusa 1888) in tap water to achieve the desired sugar concentrations.

[0225] 2. Microbial Strains:

[0226] The following commercially available microorganisms were used: a. Malolactic Bacteria: i. Chr. Hansen VINIFLORA® CH16 (Oenococcus oeni, 1.0E+11 cfu / g), 0.004g / l ii. Chr. Hansen VINIFLORA® CH35 (Oenococcus oeni, 1.0E+11 cfu / g), 0.008g / l iii. Chr. Hansen VINIFLORA® Oenos 2.0 (Oenococcus oeni, 1.0E+11 cfu / g), 0.0024g / l b. Yeast: i. Chr. Hansen VINIFLORA® FROOTZEN® (Pichia kluyveri), 0.11g / l These were used throughout the examples.

[0227] 3. Control: a. Control samples without the addition of any malolactic bacteria or yeast, set at diluted to 10 °Brix. These control samples were used as a baseline for comparison.

[0228] 4. Temperature Control: a. Room temperature conditions were maintained at 27°C throughout the experiment to ensure consistent environmental conditions for the samples, even though these were not ideal temps for testing, the room conditions did not allow for temperature variation.

[0229] 5. Laboratory Glassware and Equipment: a. Sterile plastic 4L containers for the preparation of sugar solutions and small 0.1L plastic containers for the inoculation of microbial strains. b. Sterile pipettes and pipette tips for precise measurement and handling of samples. c. Laboratory refrigerators and freezers for storage of microbial strains. d. pH meter and refractometer for monitoring and controlling the pH and sugar levels of the samples.

[0230] 6. Analytical Tools: a. Standardized sensory evaluation forms for scoring and recording sensory attributes. The materials mentioned above were utilized in the study to assess the organoleptic properties of juice solutions of varying sugar concentrations and different microbial strains, enabling the selection of the most suitable conditions for further experimentation.

[0231] Methods:

[0232] Multiple variants of sugar concentrations (5, 10, 15, 20° Brix) were prepared using dilution of 65 °Brix grape concentrate with water to reach the desired °Brix for experiment testing. The addition of different malolactic bacterial strains (CH35, CH16, Oenos 2.0) and yeast (Frootzen) from Chr. Hansen were prepared according to the instructions from the manufacturer (re-hydration of bacteria in water prior to the addition to the prepared °Brix solutions of grape juice) and a control (no bacteria, set at 10°Brix) were tested to determine which could provide the best organoleptic properties to experiment on further. Room temperature was 27°C. Samples were collected on differing days and the organoleptic properties were analyzed based on a standardized sensory evaluation (including taste, aroma, acidity and visual properties) and assigned a score between 0-5 depending on the development characteristics.

[0233] Alcohol content was measured using a spectrophotometer with enzyme kits from DNAPhone.

[0234] Results:

[0235] Table 1 shows the scores displayed from 0-5, (0 being no development / terrible organoleptic properties and 5 being great development / excellent organoleptic profile. A score of 2.5 and above is considered an acceptable result for final product, but the highest score is always preferred). It was observed that 5 and 20°Brix provided the worst results, so it was decided to continue further with 10 and 15°Brix.

[0236] Table 1

[0237] Example 2: Temperature

[0238] Method

[0239] Materials:

[0240] 1. Sugar Concentrations : a. Juice solutions with multiple variants of sugar concentrations: 10 °Brix and 15 °Brix. These solutions were prepared by dissolving 65 °Brix grape concentrate (Brusa 1888) in reverse osmosis water to achieve the desired sugar concentrations.

[0241] 2. Microbial Strains: a. Malolactic Bacteria: i.Chr. Hansen VINIFLORA® CH16 (Oenococcus oeni, 1.0E+11 cfu / g), 0.004g / l ii.Chr. Hansen VINIFLORA® CH35 (Oenococcus oeni, 1.0E+11 cfu / g), 0.008g / l iii.Chr. Hansen VINIFLORA® Oenos 2.0 (Oenococcus oeni, 1.0E+11 cfu / g), 0.0024g / l iv.Chr. Hansen VINIFLORA® Nova Plus (Lactobacillus plantarum, 2.0E+11 cfu / g), 0.002 g / i

[0242] 3. Temperature Control: a. Two temperature variants were tested in 2 different locations. Room temperature conditions were maintained at 27°C in one location and 22°C in another room throughout the experiment to ensure consistent environmental conditions for the samples. The room conditions did not allow for temperature variation.

[0243] 4. Laboratory Glassware and Equipment: a. Sterile plastic 4L containers for the preparation of sugar solutions and small 0.1L plastic containers for the inoculation of microbial strains. b. Sterile pipettes and pipette tips for precise measurement and handling of samples. c. Laboratory refrigerators and freezers for storage of microbial strains. d. pH meter and refractometer for monitoring and controlling the pH and sugar levels of the samples.

[0244] 5. Analytical Tools: a. Standardized sensory evaluation forms for scoring and recording sensory attributes.

[0245] 6. Chemicals and Reagents: a. Reverse osmosis water for the preparation of juice solutions.

[0246] The materials mentioned above were utilized in the study to assess the organoleptic properties of juice solutions of varying sugar concentrations and different microbial strains, enabling the selection of the most suitable conditions for further experimentation.

[0247] Methods:

[0248] Multiple variants of sugar concentrations (10, 15° Brix) were prepared using dilution of 65 °Brix grape concentrate with reverse osmosis water to reach the desired °Brix for experiment testing. The addition of different malolactic bacterial strains (CH35, CH16, Oenos 2.0, Nova Plus) from Chr. Hansen were prepared according to the instructions from the manufacturer (re-hydration of bacteria in water prior to the addition to the prepared °Brix solutions of grape juice) were tested at differing room temperatures (22°C and 27°C) to determine which could provide the best organoleptic properties to experiment on further and if temperature is a determining factor. Samples were collected on differing days and the organoleptic properties were analyzed based on a standardized sensory evaluation (including taste, aroma, acidity, and visual properties) used for all examples, and assigned a score between 0-5 depending on the development characteristics.

[0249] Results:

[0250] Displayed from 0-5, (0 being no development / terrible organoleptic properties and 5 being great development / excellent organoleptic profile. Scores of 2.5 and above are considered an acceptable result, however the highest score is always preferred). It was determined that temperature plays an important role (22°C creating a superior base) and that CH35 and Nova Plus were the superior bacteria in a 10° Brix solution at 18 days and 10 days respectively. Upon alcohol testing it was determined that Day 18 10° Brix CH35 produced 7.42% ale. and Day 12 10° Brix Nova Plus produced 4.19% ale. Table 2a

[0251] Table 2b Table 2c Table 2d

[0252] Example 3: Effect of ethanol production inhibitor / sequestrant

[0253] Materials:

[0254] 1. Sugar Concentrations: a. Juice solutions with multiple variants of sugar concentrations: 10 °Brix and 15°Brix prepared by dissolving 65 °Brix grape concentrate (Brusa 1888) in reverse osmosis water to achieve the desired sugar concentrations.

[0255] 2. Microbial Strains: a. Malolactic Bacteria: i.Chr. Hansen VINIFLORA® CH35 (Oenococcus oeni, 1.0E+11 cfu / g), 0.008g / l ii.Chr. Hansen VINIFLORA® Nova Plus (Lactobacillus plantarum, 2.0E+11 cfu / g), 0.002 g / i

[0256] 3. Temperature Control: a. Room temperature conditions were maintained at 22°C throughout the experiment to ensure consistent environmental conditions for the samples.

[0257] 4. Laboratory Glassware and Equipment: a. Sterile 5L glass demijohns with airlocks for the preparation of diluted juice solutions and small 0.1L plastic containers for the inoculation of microbial strains. b. Sterile pipettes and pipette tips for precise measurement and handling of samples. c. Laboratory refrigerators and freezers for storage of microbial strains. 5. Analytical Tools: a. Standardized sensory evaluation forms for scoring and recording sensory attributes. b. pH meter and refractometer for monitoring and controlling the pH and sugar levels of the samples.

[0258] 6. Chemicals and Reagents: a. Reverse osmosis water for the preparation of juice solutions. b. Potassium Sorbate (CAS Number: 24634-61-5, Vinoferm SORBISTAT)

[0259] The materials mentioned above were utilized in the study to assess the organoleptic properties of juice solutions of varying sugar concentrations and different microbial strains, enabling the selection of the most suitable conditions for further experimentation. Furthermore, the addition of potassium sorbate was tested to see if it could inhibit the formation of alcohol during fermentation.

[0260] Methods:

[0261] Multiple variants of sugar concentrations (10, 15° Brix) were prepared in a 5L demijohn using dilution of 65 °Brix grape concentrate with reverse osmosis water to reach the desired °Brix for experiment testing. The addition of different malolactic bacterial strains (CH35 and Nova Plus) from Chr. Hansen were prepared according to the instructions from the manufacturer (re-hydration of bacteria in water prior to the addition to the prepared °Brix solutions of grape juice). The addition of Potassium Sorbate at a dose of 0.2g / l was added to test its inhibiting properties on the formation of alcohol during malolactic fermentation. Potassium sorbate was added directly into the prepared solution and mixed thoroughly. The experiment was conducted at a room temperature of 22°C. Samples were collected on differing days and the organoleptic properties were analyzed based on a standardized sensory evaluation (including taste, aroma, acidity and visual properties) and assigned a score between 0-5 depending on the development characteristics.

[0262] Results:

[0263] It was determined that the addition of Potassium Sorbate inhibited the formation of alcohol during fermentation and that Nova Plus produced the best organoleptic properties in the shortest amount of time. Only 1 trial for Nova Plus is shown below; however, the effects of Potassium Sorbate on alcohol formation were seen throughout multiple trials. The table 3 below shows organoleptic properties scored from 0-5, (0 being no development / terrible organoleptic properties and 5 being great development / excellent organoleptic profile. >2.5 is considered an acceptable result, however the highest score is always preferred). Table 3- Sample

[0264] Conclusion

[0265] The results show that the non-alcoholic beverage (wine analogue) had distinct flavour profile, reminiscent of alcoholic wine in in particular in the flavour and sweetness.

[0266] Furthermore, as is evident from the table above, it is highly important to determine the optimal duration of fermentation and stop the fermentation process when the organoleptic properties and alcohol percentage are optimal / desired. In this case, the best outcome would have been obtained from stopping the fermentation before day 12 as the organoleptic properties drop from day 10 to day 12 and as the alcohol content rises to a level which is too high (above 0.5 % ABV) when fermenting beyond day 10.

[0267] Example 4: Testing 7 and 10° Brix with Nutrients and Oak

[0268] Materials:

[0269] 1. Sugar Concentrations: a. Juice solutions with multiple variants of sugar concentrations: 7 °Brix 10

[0270] °Brix prepared by dissolving 65 °Brix grape concentrate (Brusa 1888) in reverse osmosis water to achieve the desired sugar concentrations.

[0271] 2. Microbial Strains: a. Malolactic Bacteria: i.Chr. Hansen VINIFLORA® CH35 (Oenococcus oeni, 1.0E+11 cfu / g), 0.008g / l ii.Chr. Hansen VINIFLORA® Nova Plus (Lactobacillus plantarum, 2.0E+11 cfu / g), 0.002 g / i

[0272] 3. Temperature Control: a. Room temperature conditions were maintained at 21°C throughout the experiment to ensure consistent environmental conditions for the samples.

[0273] 4. Laboratory Glassware and Equipment: a. Sterile 5L glass demijohns with airlocks for the preparation of diluted juice solutions and small 0.1L plastic containers for the inoculation of microbial strains. b. Sterile pipettes and pipette tips for precise measurement and handling of samples. c. Laboratory refrigerators and freezers for storage of microbial strains.

[0274] 5. Analytical Tools: a. Standardized sensory evaluation forms for scoring and recording sensory attributes. b. pH meter and refractometer for monitoring and controlling the pH and sugar levels of the samples.

[0275] 6. Chemicals and Reagents and Miscellaneous: a. Reverse osmosis water for the preparation of juice solutions. b. Potassium Sorbate (CAS Number: 24634-61-5, Vinoferm SORBISTAT) c. Enartis Nutriferm ML (0.25g / L) d. Enartis Nutriferm Osmobacti (0.02g / L) e. Brewferm oak chips American Heavy Toast (2g / L) f. Brewferm oak chips American Medium Toast (2g / L)

[0276] The materials mentioned above were utilized in the study to assess the organoleptic properties of juice solutions of varying sugar concentrations and different microbial strains, enabling the selection of the most suitable conditions for further experimentation. Furthermore, the addition of potassium sorbate, oak chips as well as bacterial nutrients were tested to see their impact on the starting substrate.

[0277] Methods: Multiple variants of sugar concentrations (7, 10 °Brix) were prepared in a 5L demijohn using dilution of 65 °Brix grape concentrate with reverse osmosis water to reach the desired °Brix for experiment testing. The addition of different malolactic bacterial strains (CH35 and Nova Plus) from Chr. Hansen were prepared according to the instructions from the manufacturer (re-hydration of bacteria in water prior to the addition to the prepared °Brix solutions of grape juice). Enartis Nutriferm Osmobacti was added during the rehydration process at a dose of 0.02g / l of the final juice quantity prepared. The addition of Potassium Sorbate at a dose of 0.2g / l and Enartis Nutriferm ML at a dose of 0.25g / l were added directly into the prepared juice solution and mixed thoroughly. Differing oak chips were also tested to see the impact on organoleptic developments (Brewferm oak chips American Heavy Toast & Brewferm oak chips American Medium Toast) at a dose of 2g / l of final juice substrate prepared. The experiment was conducted at a room temperature of 21°C. Samples were collected on differing days and the organoleptic properties were analyzed based on a standardized sensory evaluation (including taste, aroma, acidity and visual properties) and assigned a score between 0-5 depending on the development characteristics.

[0278] Results:

[0279] It was determined that the addition of nutrients created ideal organoleptic properties with a shorter fermentation than control. Nova Plus 7° Brix was determined to contain the best organoleptic properties of the experiments (results only displayed for Nova Plus). Displayed from 0-5, (0 being no development / terrible organoleptic properties and 5 being great development / excellent organoleptic profile. >2.5 is considered an acceptable result, however the highest score is always preferred).

[0280] Table 4

[0281] Example 5: Final °Brix Determination and Bacteria Selection

[0282] Materials

[0283] 1. Sugar Concentrations: a. Juice solutions with multiple variants of sugar concentrations: 6.5, 7, 7.5 and 8.5 °Brix prepared by dissolving 65 °Brix grape concentrate (Brusa 1888) in reverse osmosis water to achieve the desired sugar concentrations.

[0284] 2. Microbial Strains: a. Malolactic Bacteria: i.Chr. Hansen VINIFLORA® CH35 (Oenococcus oeni, 1.0E+11 cfu / g), 0.008g / l ii.Chr. Hansen VINIFLORA® Nova Plus (Lactobacillus plantarum, 2.0E+11 cfu / g), 0.002 g / i

[0285] 3. Temperature Control: a. Room temperature conditions were maintained at 21°C throughout the experiment to ensure consistent environmental conditions for the samples.

[0286] 4. Laboratory Glassware and Equipment: a. Sterile 5L glass demijohns with airlocks for the preparation of diluted juice solutions and small 0.1L plastic containers for the inoculation of microbial strains. b. Sterile pipettes and pipette tips for precise measurement and handling of samples. c. Laboratory refrigerators and freezers for storage of microbial strains.

[0287] 5. Analytical Tools: a. Standardized sensory evaluation forms for scoring and recording sensory attributes. b. pH meter and refractometer for monitoring and controlling the pH and sugar levels of the samples.

[0288] 6. Chemicals and Reagents and Miscellaneous: a. Deionized water for the preparation of juice solutions. b. Potassium Sorbate (CAS Number: 24634-61-5, Vinoferm SORBISTAT) c. Enartis Nutriferm ML (0.25g / L) d. Enartis Nutriferm Osmobacti (0.02g / L) e. Brewferm oak chips American Heavy Toast (2g / L)

[0289] The materials mentioned above were utilized in the study to assess the organoleptic properties of juice solutions of varying sugar concentrations and different microbial strains, enabling the selection of the most suitable conditions for further experimentation. Furthermore, the addition of potassium sorbate, oak chips as well as bacterial nutrients were tested to see their impact on the starting substrate.

[0290] Method

[0291] Multiple variants of sugar concentrations (6.5, 7, 7.5 and 8.5 °Brix) were prepared in a 5L demijohn using dilution of 65 °Brix grape concentrate with de-mineralized water to reach the desired °Brix for experiment testing. The addition of different malolactic bacterial strains (CH35 and Nova Plus) from Chr. Hansen were prepared according to the instructions from the manufacturer (re-hydration of bacteria in water prior to the addition to the prepared °Brix solutions of grape juice). Enartis Nutriferm Osmobacti was added during the rehydration process at a dose of 0.02g / l of the final juice quantity prepared. The addition of Potassium Sorbate at a dose of 0.2g / l and Enartis Nutriferm ML at a dose of 0.25g / l was added directly into the prepared juice solution and mixed thoroughly. Brewferm oak chips American Heavy Toast were used at a dose of 2g / l of final juice substrate prepared. The experiment was conducted at a room temperature of 21°C. Samples were collected on differing days and the organoleptic properties were analyzed based on a standardized sensory evaluation (including taste, aroma, acidity and visual properties) and assigned a score between 0-5 depending on the development characteristics.

[0292] Result

[0293] It was determined that the base starting concentrations of 7-7.5 °Brix in a substrate with Nova Plus produced the best organoleptic properties with a low sweetness perception to be used as a starting base for recipe creation (results only displayed for Nova Plus). Displayed from 0-5, (0 being no development / terrible organoleptic properties and 5 being great development / excellent organoleptic profile. >2.5 is considered an acceptable result, however the highest score is always preferred). (Experiments were conducted at different times hence different collection dates). Table 5

[0294] Example 6: Miscellaneous

[0295] Materials 1. Sugar Concentrations: a. Juice solutions with sugar concentrations of 7 °Brix prepared by dissolving 65 °Brix grape concentrate (Brusa 1888) in reverse osmosis water to achieve the desired sugar concentrations. Chardonnay Juice (Beutelsbacher) was also diluted with de-mineralized water to attain a 7 °Brix preparation. 2. Microbial Strains: a. Malolactic Bacteria: i.Chr. Hansen VINIFLORA® CH35 (Oenococcus oeni, 1.0E+11 cfu / g), 0.008g / l ii.Chr. Hansen VINIFLORA® CH16 (Oenococcus oeni, 1.0E+11 cfu / g), 0.004g / l iii.Chr. Hansen VINIFLORA® Oenos 2.0 (Oenococcus oeni, 1.0E+11 cfu / g), 0.0024g / l iv.Chr. Hansen VINIFLORA® Nova Plus (Lactobacillus plantarum, 2.0E+11 cfu / g), 0.002 g / i

[0296] 3. Temperature Control: a. Room temperature conditions were maintained at 21°C throughout the experiment to ensure consistent environmental conditions for the samples.

[0297] 4. Laboratory Glassware and Equipment: a. Sterile 5L glass demijohns with airlocks for the preparation of diluted juice solutions and small 0.1L plastic containers for the inoculation of microbial strains. b. Sterile pipettes and pipette tips for precise measurement and handling of samples. c. Laboratory refrigerators and freezers for storage of microbial strains.

[0298] 5. Analytical Tools: a. Standardized sensory evaluation forms for scoring and recording sensory attributes. b. pH meter and refractometer for monitoring and controlling the pH and sugar levels of the samples.

[0299] 6. Chemicals and Reagents and Miscellaneous: a. Reverse osmosis water for the preparation of juice solutions. b. Potassium Sorbate (CAS Number: 24634-61-5, Vinoferm SORBISTAT) c. Enartis Nutriferm ML (0.25g / L) d. Enartis Nutriferm Osmobacti (0.02g / L)

[0300] The materials mentioned above were utilized in the study to assess the organoleptic properties of juice solutions of 7 °Brix concentrations with sequential inoculations of different microbial strains, to test whether the use of different bacteria and sequential inoculation could result in better organoleptic properties of a starting base. Furthermore, specific grape variety juice was also tested to see if similar organoleptic developments can occur with a different starting substrate.

[0301] Methods:

[0302] Multiple 7 °Brix solutions were prepared in a 5L demijohn using dilution of 65 °Brix grape concentrate with reverse osmosis water to reach the desired 7 °Brix for experiment testing. One sample was created using 16.75 °Brix Chardonnay juice diluted with reverse osmosis water to reach the desired 7 °Brix for experiment testing, this experiment was inoculated with Chr. Hansen VINIFLORA® Nova Plus which was prepared according to instructions from the manufacturer (re-hydration of bacteria in water prior to the addition to the prepared °Brix solutions of grape juice). The addition of different malolactic bacterial strains (CH35, Nova Plus, CH16 and Oenos 2.0) from Chr. Hansen were prepared according to the instructions from the manufacturer on the day of their inoculation (re-hydration of bacteria in water prior to the addition to the prepared °Brix solutions of grape juice). Enartis Nutriferm Osmobacti was added during the rehydration process at a dose of 0.02g / l of the final juice quantity prepared. The addition of Potassium Sorbate at a dose of 0.2g / l and Enartis Nutriferm ML at a dose of 0.25g / l was added directly into the prepared juice solution and mixed thoroughly. The experiment was conducted at a room temperature of 21°C. Samples were collected on differing days and the organoleptic properties were analyzed based on a standardized sensory evaluation (including taste, aroma, acidity and visual properties) and assigned a score between 0-5 depending on the development characteristics. The differing experiments were prepared according to the protocol above and differed only in bacterial strains and inoculation days. The experiment designs were as follows: Experiment 1 inoculation on Day 1 with Chr. Hansen VINIFLORA® Nova Plus which was followed by a sequential inoculation at Day 3 with Chr. Hansen VINIFLORA® Oenos 2.0, Experiment 2 inoculation on Day 1 with Chr. Hansen VINIFLORA® Nova Plus which was followed by a sequential inoculation at Day 3 with Chr. Hansen VINIFLORA® CH35, Experiment 3 inoculation on Day 1 with Chr. Hansen VINIFLORA® Oenos 2.0 which was followed by a sequential inoculation at Day 3 with Chr. Hansen VINIFLORA® Nova Plus, and Experiment 4 inoculation on Day 1 with Chr. Hansen VINIFLORA® Nova Plus which was followed by a sequential inoculation at Day 3 with Chr. Hansen VINIFLORA® CH35 and another sequential inoculation at Day 6 with Chr. Hansen VINIFLORA® Oenos 2.0.

[0303] Results:

[0304] It was determined that Experiment 1 and 2 had the best organoleptic outcomes after fermentation, however fermentation with only Nova Plus (original developed method) still possesses the best properties (see Example 5). Fermentations using diluted variety grape juice also shows potential as a starting base substrate. Displayed from 0-5, (0 being no development / terrible organoleptic properties and 5 being great development / excellent organoleptic profile. >2.5 is considered an acceptable result, however the highest score is always preferred). See also Figures 1, 2 and 3.

[0305] The final Organoleptic Characteristics Post Fermentation were: Aroma : Rhubarb, citrus, buttery, light vanilla and oak Flavour: Citrus (lemon) dominant, light rhubarb and oak notes, slight vanilla, caramel, raspberry with a nice acidic balance and slight tannic mouthfeel pH: 2.9-3.2 °Brix: 6.8-7.5 (depending on product variety) Table 4

[0306] Example 7: Further non-alcoholic beverages Using the same materials, parameters and methods as described in example 6, unless otherwise stated, a number of non-alcoholic beverages NBC01-NBC44 were made. Certain key materials are noted in the below tables 5-9, some of which represent deviations from example 6. Table 5

[0307] Oenococcus oeni was provided as CH35 as in example 6.

[0308] Table 6

[0309] Oenococcus oeni was provided as CH35 as in example 6, whereas Lactobacillus plantarum was provided as Nova Plus as in example 6.

[0310] Table 7

[0311] Oenococcus oeni was provided as CH35 as in example 6.

[0312] Table 8

[0313] Oenococcus oeni was provided as CH35 as in example 6.

[0314] Table 9

[0315] Oenococcus oeni was provided as CH35 as in example 6.

[0316] Example 8: Conclusion The beverages NBC01-NBC44 are acceptable as a non-alcohol beverage by having an alcohol content below 0.5% ABV.

Claims

CLAIMS1. A process for producing a non-alcoholic beverage comprising the steps of a. Providing a starting material comprising grape extract; b. Adding at least one ethanol production inhibitor, c. Inoculating the starting material with at least one malolactic bacterial strain; d. Fermenting the starting material including the at least one malolactic bacterial strain, thereby achieving a non-alcoholic beverage having an alcohol content of not more than 0.5 % Alcohol By Volume (ABV).

2. The process according to claim 1, wherein step b is performed prior to step d.

3. The process according to claim 1 or 2, wherein step d comprises fermenting the starting material including the at least one malolactic bacterial strain and the at least one ethanol production inhibitor,4. The process according to any of the preceding claims, wherein step b comprises adding the at least one ethanol production inhibitor to the starting material having a pH value of at least 3.0 at the time of addition, such as at least 3.5 at the time of addition, such as a pH value of at least 4.0 at the time of addition.

5. The process according to any of the preceding claims, wherein step d results in a change of pH value, wherein step b occurs when no more than 50 % of the change of pH value has occurred, such as no more than 20 % of the change of pH value has occurred.

6. The process according to any of the preceding claims, wherein step d results in a change of brix value, wherein step b occurs when no more than 50% of the change of brix value has occurred, such as no more than 20% of the change of brix value has occurred.

7. The process according to any of the preceding claims, wherein step b comprises adding the at least one ethanol production inhibitor to the fermentation mixture havingan alcohol content of no more than 8.0 % ABV, such as no more than 3.0% ABV, such as no more than 1.0 % ABV, such as no more than 0.5 % ABV.

8. The process according to any of the preceding claims, wherein the fermentation is performed for a duration of which at least 50% is after step b, such as at least 80% is after step b, such as at least 90% is after step b, such as at least 95% is after step b, such as at least 98% is after step b, such as at least 99% is after step b.

9. The process according to any of the preceding claims, wherein step d results in a change in cfu count, wherein step b occurs when no more than 50% of the change in the cfu count has occurred, such as no more than 20% of the change in the cfu count has occurred.

10. The process according to any of the preceding claims, wherein the process comprises a primary fermentation and optionally a secondary fermentation, and wherein the ethanol production inhibitor is added before initiation of the primary fermentation.

11. The process according to any of the preceding claims, wherein the grape extract is made from one or more grape varieties selected from the group consisting of Chardonnay, Sauvignon Blanc, Pinot Gris, Pinot Noir, Semilion, Gewurztraminer, Chenin Blanc, Pinot Blanc, Viognier, Roussanne, Marsanne, Muscat, Riesling, Silvaner, Muller-Thurgau, Gruner Veltliner, Torrontes, Garganega, Verdicchio, Sangiovese, Tempranillo, Zinfandel, Pinotage, Cabernet Franc, Merlot, Albarino, Airen, and any combination thereof.

12. The process according to any of the preceding claims, wherein fermenting the starting material is performed by the malolactic bacteria.

13. The process according to any of the preceding claims, wherein fermenting the starting material is a primary fermentation.

14. The process according to any of the preceding claims, wherein fermenting the starting material is a primary fermentation performed by the malolactic bacteria.

15. The process according to any of the preceding claims, wherein no secondary fermentation is performed.

16. The process according to any of the preceding claims, wherein a fermentation mixture is obtained from the starting material and the at least one bacterial culture.

17. The process according to any of the preceding claims, wherein the viable yeast count in the fermentation mixture is equal to or below below 3.0*10 4 cfu / mL, such as below 2.0*10 4 cfu / mL, such as below 1.56*10 4 cfu / mL, such as below 1.0*10 4 cfu / mL, such as below 3.0*10 3 cfu / mL, such as below 100 cfu / mL, such as below 10 cfu / mL 1.56*10 4 cfu / mL..

18. The process according to any of the preceding claims, wherein step b is performed when the cfu count of ethanol-producing microorganisms in the starting material is between 0 and 1.563.0*10^4 cfu / mL,. such as between 0 and 2.0*10 4 cfu / mL, such as between 0 and 1.56*10 4 cfu / mL, such as between 0 and 1.0*10 4 cfu / mL, such as between 0 and 3.0*10 3 cfu / mL, such as between 0 and 100 cfu / mL, such as between 0 and 10 cfu / mL.

19. The process according to any of the preceding claims, wherein the viable yeast count in the fermentation mixture is equal to or below 5 cfu / mL.

20. The process according to any of the preceding claims, wherein the inoculation is performed once.

21. The process according to any of the preceding claims, wherein the inoculation is a bacterial inoculation.

22. The process according to any of the preceding claims, wherein the inoculation is performed once and wherein the inoculation is a bacterial inoculation.

23. The process according to any of the preceding claims, wherein no yeast is added.

24. The process according to any of the preceding claims, wherein no viable yeast is added.

25. The process according to any of the preceding claims, wherein no viable Saccharomyces yeast is added.

26. The process according to any of the preceding claims, wherein the at least one bacterial strain comprises a malolactic bacterial strain, such as consists of a malolactic bacterial strain.

27. The process according to any of the preceding claims, wherein the at least one malolactic bacterial strain includes lactic acid bacteria.

28. The process according to any of the preceding claims, wherein the at least one bacterial strain is selected from the group consisting of Lactobacillus, Leuconostoc, Pediococcus, Lactococcus, and Streptococcus, Aerococcus, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Tetragenococcus, Vagococcus Weissella, and any combination thereof.

29. The process according to any of the preceding claims, wherein the at least one malolactic bacterial strain is selected from the group consisting of one or more Lactobacillus plantarum ssp and / or one or more Oenococcus oeni ssp.

30. The process according to any of the preceding claims, wherein the at least one bacterial strain is Lactobacillus plantarum ssp.

31. The process according to any of the preceding claims, wherein the at least one bacterial strain is Oenococcus oeni ssp.

32. The process according to any of the preceding claims, wherein the at least one bacterial strain comprises a non-malolactic bacterial strain, such as a non-malolactic bacterial strain selected from the group consisting of Actinobacillus succinogenes, Anaerobiospirillum succiniciproducens, Escherichia coli, Acetobacteraceae (e.g. acetobacter and gluconobacter), Pseudomonadaceae, Enterobacteriaceae, and any combination thereof.

33. The process according to any of the preceding claims, wherein the at least one malolactic bacterial strain is added in an amount in the range of from 2.0*10 8 to 1.5*10 10 cfu / L starting material.

34. The process according to any of the preceding claims, wherein the starting material has a Brix value in the range from 0.5 to 75.

35. The process according to any of the preceding claims, wherein the starting material has a Brix value in the range from 2 to 20.

36. The process according to any of the preceding claims, wherein the starting material has a Brix value in the range from 5 to 10.

37. The process according to any of the preceding claims, wherein the Brix value of the non-alcoholic beverage is in the range from 0.25 to 30.

38. The process according to any of the preceding claims, wherein the Brix value of the non-alcoholic beverage is in the range from 2 to 7.

39. The process according to any of the preceding claims, wherein the process further comprises a step of adding nutrients to the fermentation mixture.

40. The process according to any of the preceding claims, wherein the process further comprises a step of adding stability enhancers to the fermentation mixture.

41. The process according to any of the preceding claims, wherein the process further comprises a step of adding regulators of osmotic pressure to the bacteria before inoculation.

42. The process according to any of the preceding claims, wherein fermenting the starting material comprises conversion of malic acid into lactic acid and carbon dioxide.

43. The process according to any of the preceding claims, wherein fermenting the starting material does not produce alcohol.

44. The process according to any of the preceding claims, wherein fermenting the starting material is anaerobic.

45. The process according to any of the preceding claims, wherein the fermentation is performed for a duration which is no more than 15 days.

46. The process according to any of the preceding claims, wherein the fermentation is performed for a duration which is no more than 45 days.

47. The process according to any of the preceding claims, wherein fermenting the starting material is performed for a duration of 1 hour to 45 days, such as 2 hours to 20 days, such as 3 hours to 15 days, such as 4 hours to 10 days.

48. The process according to any of the preceding claims, wherein the ethanol production inhibitor comprises one or more selected from the group consisting of sorbic acid, sorbic acid salts, benzoic acid, benzoic acid salts, and any combination thereof.

49. The process according to any of the preceding claims, wherein the ethanol production inhibitor comprises one or more selected from the group consisting of sorbic acid, potassium sorbate, benzoic acid, sodium benzoate, potassium benzoate, calcium benzoate, ethyl-p-hydroxybenzoate, sodium ethyl p-hydroxybenzoate, methyl p- hydroxybenzoate, sodium methyl p-hydroxybenzoate, and any combination thereof.

50. The process according to any of the preceding claims, wherein the ethanol production inhibitor comprises potassium sorbate and / or sodium benzoate.

51. The process according to any of the preceding claims, wherein the ethanol production inhibitor is a yeast fermentation inhibitor.

52. The process according to any of the preceding claims, wherein the ethanol production inhibitor is added in an amount of between 0.01 and 5 g / L, such as between 0.01 and 1 g / L.

53. The process according to any of the preceding claims, wherein the ethanol production inhibitor is added in an amount of between 0.1 and 0.5 g / L.

54. The process according to any of the preceding claims, wherein the process further comprises adding fermentation aids and / or flavouring ingredients.

55. The process according to any of the preceding claims, wherein the flavouring ingredients include oak chips.

56. The process according to any of the preceding claims wherein, fermentation aids and / or flavouring agents are added before or during fermentation.

57. The process according to any of the preceding claims, wherein the flavouring ingredients include tannins.

58. The process according to any of the preceding claims, wherein the flavouring ingredients comprise botanical flavour ingredients and / or natural aromas.

59. The process according to claim 58, wherein the botanical flavour ingredients and / or natural aromas are derived from fruit, berries, herbs, spices, aromatic plant parts, or any combination thereof.

60. The process according to any of the preceding claims, wherein said flavouring ingredients and / or natural aromas comprise one or more sweeteners.

61. The process according to any of the preceding claims, wherein the process further comprises a step of fining the fermentation mixture after fermentation.

62. The process according to claim 61, wherein the step of fining the fermentation mixture includes the addition of fining agents such as organic fining components or such as mineral fining components.

63. The process according to claim 62, wherein the fining agents comprise Kieselsol and / or Chitosan.

64. The process according to claim 62 or 63, wherein the organic fining components comprise egg whites, casein, gelatine, chitin, isinglass, and any combination thereof.

65. The process according to any of claims 62-64, wherein the mineral fining components comprise bentonite, charcoal, potassium ferrocyanide, silica, kaolin, and any combination thereof.

66. The process according to any of the preceding claims, wherein the process further comprises a step of sterilizing the fermentation mixture after fermentation.

67. The process according to claim 66, wherein sterilizing the fermentation mixture comprises sterile filtration and / or pasteurization.

68. The process according to any of the preceding claims, wherein the process further comprises adding sulfites to the fermentation mixture after fermentation.

69. The process according to any of the preceding claims, wherein the process further comprising bottling, canning, boxing or kegging the non-alcoholic beverage.

70. The process according to any of the preceding claims, wherein the process further comprises a step of filtering the fermentation mixture after fermentation.

71. The process according to any of the preceding claims, wherein filtering the fermentation mixture after fermentation comprises a first filtration step and a second filtration step.

72. The process according to claim 71, wherein the first filtration step is performed using a filter having a pore size of at least 2 micron.

73. The process according to claim 71 or 72, wherein the second filtration step is performed using a filter having a pore size between 0.1 and 2.0 micrometers such as between 0.2 and 1.5 micrometers, such as between 0.3 and 1.0 micrometers, such as between 0.1 and 0.5 micrometers, such as between 0.2 and 0.5 micrometers, such as between 0.3 and 0.5 micrometers, such as between 0.4 and 0.6 micrometres.

74. The process according to any of the preceding claims, wherein the starting material has been subject to enzymatic treatment prior to inoculating the starting material with at least one bacterial strain.

75. The process according to any of the preceding claims, wherein the starting material is fermented until the pH is in the range of from 2 to 4.

76. The process according to any of the preceding claims, wherein the process further comprises a step of adding nutrients and / or growth support agents to the fermentation mixture.

77. The process according to claim 76, wherein the nutrients and / or growth support agents comprise inactivated yeast, protein hydrolysate, amino acids, polysaccharides, bacterial cell wall polypeptides, parietal polysaccharides, cofactors, vitamins, and any combination thereof.

78. The process according to claim 76 or 77, wherein the nutrients and / or growth support agents comprise Enartis Nutriferm ML and / or Enartis Nutriferm Osmobacti.

79. The process according to any of the preceding claims, wherein the non-alcoholic beverage is a non-alcoholic wine analogue.

80. A process for producing a non-alcoholic beverage comprising the steps ofa. Providing a starting material consisting of a grape extract; wherein the starting material has a Brix value in the range from 5 to 10, such as from 6 to 8; b. Adding at least one malolactic bacterial strain to the starting material; wherein said bacterial strain is selected from the group consisting of one or more Lactobacillus plantarum ssp and / or one or more Oenococcus oeni ssp; c. Fermenting the starting material comprising the at least one malolactic bacterial strain until the pH is in the range of from 2-4; d. Adding Potassium sorbate in the range from 0.1 to 1 g / L; e. Adding flavour ingredients comprising adding oak chips in an amount in the range from 0.01 to 2 g / L; f. Fining the non-alcoholic beverage; g. Filtering the non-alcoholic beverage; h. Bottling the non-alcoholic beverage; thereby achieving a non-alcoholic beverage having an alcohol content of not more than 0.5% ABV.

81. A process for producing a non-alcoholic beverage comprising the steps of a. Providing a starting material comprising grape extract; wherein the starting material has a Brix value in the range from 6 to 10; b. Adding at least one malolactic bacteria to the starting material; c. Fermenting the starting material comprising the at least one malolactic acid bacteria until the pH is in the range of from 2 to 3.5; d. Addition of a least one agent which prevents yeast fermentation, for example a sequestrant, to the fermentation; e. Optionally adding at least one flavour ingredient, thereby achieving a non-alcoholic beverage having an alcohol content of not more than 0.5% ABV.

82. The process according to any of the preceding claims, wherein the ethanol production inhibitor comprises potassium sorbate in the range from 0.1 to 1 g / L.

83. The process according to any of the preceding claims, wherein the process further comprises at least one step selected from the group consisting of- Fining the non-alcoholic beverage;- Filtering the non-alcoholic beverage;- Bottling the non-alcoholic beverage; and any combination thereof.

84. Use of the malolactic bacterial strain in a process according to any of claims 1 to84.

85. A non-alcoholic beverage obtainable or obtained by the process according to any of claims 1 to 83.

86. A non-alcoholic beverage having an alcohol content of not more than 0.5% ABV alcohol, a pH in the range of 2-4s and a Brix value of 5 to 10 °Brix.

87. A non-alcoholic beverage having an alcohol content of not more than 0.5% ABV alcohol, the beverage comprising fermented grape extract, wherein the fermented grape extract is obtained without yeast fermentation.

88. A non-alcoholic beverage having an alcohol content of not more than 0.5% ABV alcohol, the beverage comprising fermented grape extract, wherein the beverage is made without removing alcohol.

89. A non-alcoholic beverage having an alcohol content of not more than 0.5% ABV alcohol, the beverage comprising fermented grape extract and an ethanol production inhibitor of less than 0.5 g / L.

90. A non-alcoholic beverage according to any of claims 87-89 obtainable by the process according to any of the claims 1-83.

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