WO2017068017A1

WO2017068017A1 - Low sugar flavoured yogurt

Info

Publication number: WO2017068017A1
Application number: PCT/EP2016/075164
Authority: WO
Inventors: Dominik Bohdan GRABINSKI
Original assignee: Dsm Ip Assets B.V.
Priority date: 2015-10-23
Filing date: 2016-10-20
Publication date: 2017-04-27
Also published as: EP3364766A1

Abstract

The present invention describes flavoured yogurt comprising added sucrose in an amount of 10% (wt) or less and comprising one or more weakly post acidifying lactic acid bacteria and a non-sucrose sweetener. Further, the invention describes a process for the production of a flavoured yogurt, comprising fermenting milk having an amount of sucrose of 10% (wt) or less with a composition comprising a weakly post acidifying starter culture to provide the flavoured yogurt, and comprising adding a sweetener to the milk having an amount of sucrose of 10% (wt) or less or to the provided flavoured yogurt.

Description

LOW SUGAR FLAVOURED YOGURT

Field of the invention

The present invention relates to a flavoured yogurt. According to a further aspect, the present invention relates to a process for the production of a flavoured yogurt. According to yet another aspect, the present invention relates to the use of lactic acid bacteria.

Background of the invention

The food industry uses different bacteria, in the form in of ferments, in particular lactic acid bacteria, in order to improve the taste and the texture of foods but also to extend the shelf life of these foods. In the case of the dairy industry, lactic acid bacteria are used intensively in order to bring about the acidification of milk (by fermentation) but also in order to texturize the product into which they are incorporated. Among the lactic acid bacteria used in the food industry, there can be mentioned the genera Streptococcus and Lactobacillus. The lactic acid bacterial species Streptococcus thermophilus and Lactobacillus delbrueckii ssp bulgaricus are used in particular in the formulation of the ferments used for the production of fermented milks, for example yogurts.

The acidity produced in yogurt depends mainly on the acidifying activity of the yogurt culture (Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus) and therefore the amount of lactic acid produced during the milk maturation and also the residual acidity produced during cold storage. The texture is also varying during storage and participates in the final product sensorial properties. The recipe of the yogurt has also an impact on the yogurt sensorial properties by modifying the texture or the aroma perception.

Yogurts are often provided with added sugar in order to provide a flavoured yogurt having a sweetened flavour. However, besides providing a sweetened flavour, the added sugar also improves the texture and sensory attributes such as mouthfeel thickness and creaminess of the yogurt. Especially in countries where plain yogurt is perceived as acidic, added sugar amounts of 7 to 15 % (wt) to sweeten the product are reported. Examples of these flavoured yogurts having added sugar are fruit or vanilla yogurts.

In view of health concerns related to the consumption of sugar, there have been several research efforts for sugar substitutes, such as the use of sweeteners. However, a problem with sweeteners is that they do not provide the texturizing properties of sugar and may introduce off flavours to the yogurt.

Simply reducing the amount of added sugar negatively influences the texture and sensory attributes such as mouthfeel thickness and creaminess of the yogurt. This is considered as a disadvantage since texture and sensory attributes such as mouthfeel thickness and creaminess are important market characteristics for yogurts.

Therefore, there is a need in the art for a flavoured yogurt having a reduced amount of added sugar while maintaining the texture and sensory attributes of the flavoured yogurt.

Summary of the invention

The objective of the present invention, amongst other objectives, is to provide a flavoured yogurt having a reduced amount of added sugar while maintaining the sensory attributes of the flavoured yogurt.

This objective, amongst other objectives, is met by providing a flavoured yogurt according to the appended claim 1.

Specifically, this objective, amongst other objectives, is met by providing a flavoured yogurt having an amount of sugar of 10% (wt) or less comprising one or more weakly post acidifying lactic acid bacteria and a sweetener. More specifically, this objective, amongst other objectives, is met by providing a flavoured yogurt comprising added sucrose in an amount of 10% (wt) or less and comprising one or more weakly post acidifying lactic acid bacteria and a non- sucrose sweetener.

According to another aspect, the present invention relates to a process for the production of a flavoured yogurt, comprising fermenting milk having an amount of sugar of 10% (wt) or less with a composition comprising a weakly post acidifying starter culture to provide the flavoured yogurt, and comprising adding a sweetener to the milk having an amount of sugar 10% (wt) or less or to the provided flavoured yogurt.

According to yet another aspect, the present invention relates to the use of a lactic acid bacterial strain chosen from U, V, W, X, Y and Z for reducing the amount of added sugar in yogurt, preferably in flavoured yogurt, preferably for reducing the amount of added sugar with at least 30%, more preferably at least 50%, most preferably at least 70%.

Definitions

The term "milk" is intended to encompass milks from mammals and plant sources or mixtures thereof. Preferably, the milk is from a mammal source. Mammals sources of milk include, but are not limited to cow, sheep, goat, buffalo, camel, llama, mare and deer. In an embodiment, the milk is from a mammal selected from the group consisting of cow, sheep, goat, buffalo, camel, llama, mare and deer, and combinations thereof. Plant sources of milk include, but are not limited to, milk extracted from soy bean, pea, peanut, barley, rice, oat, quinoa, almond, cashew, coconut, hazelnut, hemp, sesame seed and sunflower seed. Soy bean milk is preferred. In addition, the term "milk" refers to not only whole milk, but also skim milk or any liquid component derived thereof. As used in the present specification, the term "yogurt" refers to products comprising lactic acid bacteria such as Streptococcus thermophilus and Lactobacillus delbruekii subsp. bulgaricus, but also, optionally, other microorganisms such as Lactobacillus delbruekii subsp. lactis, Bifidobacterium animalis subsp. lactis, Lactococcus lactis, Lactobacillus acidophilus and Lactobacillus casei, or any microorganism derived therefrom. The lactic acid strains other than Streptococcus thermophilus and Lactobacillus delbruekii subsp. bulgaricus, are intended to give the finished product various properties, such as the property of promoting the equilibrium of the flora.

As used herein, the term "yogurt" encompasses set yogurt, stirred yogurt, drinking yogurt, Petit Suisse, heat treated yogurt and yogurt-like products. Preferably, the yogurt is a stirred yogurt or a drinking yogurt. More preferably, the yogurt is a stirred yogurt. The term "yogurt" encompasses, but is not limited to, yogurt as defined according to French and European regulations, e.g. coagulated dairy products obtained by lactic acid fermentation by means of specific thermophilic lactic acid bacteria only (i.e. Lactobacillus delbruekii subsp. bulgaricus and Streptococcus thermophilus) which are cultured simultaneously and are found to be live in the final product in an amount of at least 10 million CFU (colony-forming unit) / g. Preferably, the yogurt is not heat-treated after fermentation. Yogurts may optionally contain added dairy raw materials (e.g. cream) or other ingredients such as sugar or sweetening agents, one or more flavouring(s), fruit, cereals, or nutritional substances, especially vitamins, minerals and fibers. Such yogurt advantageously meets the specifications for fermented milks and yogurts of the AFNOR NF 04-600 standard and/or the codex StanA-lla-1975 standard. In order to satisfy the AFNOR NF 04-600 standard, the product must not have been heated after fermentation and the dairy raw materials must represent a minimum of 70% (m/m) of the finished product.

The term 'flavoured yogurt' as used in the present specification, means a yogurt wherein sugar, preferably sucrose, has been added, during or after the production process. Added sugar means sugar added separately from the inherent lactic acid fermentation or lactose hydrolysis wherein glucose and galactose are formed. Preferably, lactose, glucose and galactose are not added sugar according to the present invention. Thus, a flavoured yogurt comprises an amount of added sugar, preferably an amount of added sucrose. Examples of a flavoured yogurt are vanilla, honey, toffee and fruit yogurts such as strawberry, cherry, blueberry, raspberry, mango and peach.

The term 'sweetener', as used in the present context, is intended to mean a sugar substitute which provides a sweet taste. The term 'non sucrose sweetener' as used in the present context means a sweetener which is not sucrose.

The term 'thickening agent' as used in the present context, is intended to mean a substance which increases the viscosity of the yogurt.

The term 'starter culture' or 'starter culture composition' (also referred to as 'starter') as used herein refers to a composition comprising one or more lactic acid bacteria, which are responsible for the acidification of the milk base. Starter cultures may be fresh (liquid), frozen or freeze-dried. Freeze dried cultures need to be regenerated before use. For the production of a fermented dairy product, the starter culture is usually added in an amount from 0.01 to 3%, preferably from 0.01 and 0.02 % by weight of the total amount of milk base.

As used herein, the term "lactic acid bacteria" (LAB) or "lactic bacteria" refers to food- grade bacteria producing lactic acid as the major metabolic end-product of carbohydrate fermentation. These bacteria are related by their common metabolic and physiological characteristics and are usually Gram positive, low-GC, acid tolerant, non- sporulating, non- respiring, rod-shaped bacilli or cocci. During the fermentation stage, the consumption of lactose by these bacteria causes the formation of lactic acid, reducing the pH and leading to the formation of a protein coagulum. These bacteria are thus responsible for the acidification of milk and for the texture of the dairy product. As used herein, the term "lactic acid bacteria" or "lactic bacteria" encompasses, but is not limited to, bacteria belonging to the genus of Lactobacillus spp., Bifidobacterium spp., Streptococcus spp., Lactococcus spp., such as Lactobacillus delbruekii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus lactis, Bifidobacterium animalis, Lactococcus lactis, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Lactobacillus acidophilus and Bifidobacterium breve. One or more lactic acid bacteria, as used in the present context, means one or more different lactic acid bacteria, meaning lactic acid bacteria of at least a different strain.

Lactic acid bacterial strain 'U', as used in the present context means the Streptococcus thermophilus strain as deposited under number CBS139996. Lactic acid bacterial strain 'V, as used in the present context means the Streptococcus thermophilus strain as deposited under number CBS140557. Lactic acid bacterial strain 'W, as used in the present context means the Streptococcus thermophilus strain as deposited under number CBS140559. Lactic acid bacterial strain 'X', as used in the present context means the Streptococcus thermophilus strain as deposited under number CBS140561 . Lactic acid bacterial strain Ύ', as used in the present context means the Streptococcus thermophilus strain as deposited under number CBS140558. Lactic acid bacterial strain 'Z', as used in the present context means the Lactobacillus delbrueckii ssp. bulgaricus strain as deposited under number CBS140562.

The term "weakly post-acidifying" or "low post-acidifying" refers to the acidification profile of a bacterium or a bacterial culture useful in the present invention. A weakly post-acidifying culture is defined as having a two-phase acidification profile, as determined by the continuous recordation of the pH as a function of time, in a Dutch milk composition comprising 1.5% (w/w) fat, 3.6% (w/w) protein, 4.6% (w/w) carbohydrates and 2% RSM powder and/or a Chinese milk composition comprising 2.9 % (w/w) protein, 3.6 % (w/w) fat, 4.7 % (w/w) carbohydrates at 42°C, comprising an initial period between inoculation and end of fermentation followed by a period at 4°C between 1 and 40 days in which the pH value does not fluctuate more than 0.3 unit. Weakly post-acidifying cultures may be selected by the follow-up of the pH of a final fermented milk base by using any method known by the skilled person. As example, a CINAC system (CINetic Acidification) may be used. In this system a pH meter is connected to a computer recorder and pH is continuously recorded as a function of time to obtain sigmoidal curves representing the acidification. During the follow-up of the pH, the milk is maintained at fermentation temperature in a thermoregulated bath.

"Lactase" or "beta-galactosidase" (EC: 3.2.1.23) is an enzyme, which can convert lactose (disaccharides) into the monosaccharides glucose and galactose. Lactose is present in dairy products and more particularly in milk, skimmed milk, cream and other milk products. The lactase provided in the present invention is preferably produced by a yeast, more preferably a Kluyveromyces strain, most preferably K. lactis or K. fragilis.

The term 'sensory attributes' as used in the present context means terms, or descriptors, to describe sensory characteristics of a product.

The term 'mouthfeel thickness', as used in the present context, is the degree in which the flavoured yogurt feels thick in the mouth, as analysed by a sensory panel.

The term 'creamy mouthfeel' or 'creaminess' as used in the present context is the full and creamy feeling in the mouth, by giving a velvety coating in the mouth, as analysed by a sensory panel.

The term 'sour flavour' as used in the present context is the degree in which the product tastes sour in comparison with the taste of citric acid.

The term 'a similar yogurt' as used in the present context is intended to mean a yogurt which is similar to the present flavoured yogurt in respect of recipe and fermentation conditions, however with the sole difference of a sugar content of 7%. In the event that a lactase enzyme and / or a steviol glycoside has been added to the present flavoured yogurt, the term 'a similar yogurt' refers to a yogurt which is similar to the present flavoured yogurt in respect of recipe and fermentation conditions, however with the sole difference of a sugar content, lactase enzyme and / or steviol glycoside.

The term 'comparable' as used in the present context is intended to mean as having no significant differences, as perceived by a sensory panel and / or as measured by an analysis tool such as a viscometer or a rheometer.

Weight percentages, or abbreviated as % (wt), as used in the present context means the percentage in weight, of the total weight (w/w) of the total, i.e. total milk or total yogurt.

Detailed description of the invention

According to a first aspect, the present invention relates to a flavoured yogurt comprising added sucrose in an amount of 10% (wt) or less and comprising one or more weakly post acidifying lactic acid bacteria and a non-sucrose sweetener. Alternatively, the present invention relates to a flavoured yogurt, or yogurt, having an amount of sugar of 10% (wt) or less comprising one or more weakly post acidifying lactic acid bacteria and a sweetener. The inventors of the present invention found that weakly post acidifying lactic acid bacteria are able to compensate for the loss in texture which is expected as a result of reducing the amount of sugar. Surprisingly, the flavoured yogurt of the invention has texture related sensory attributes scoring comparable with yogurts having higher amounts of sugar. For example, the present inventors found that the perceived thickness of the yogurt structure, i.e. the force needed for stirring the yogurt, is comparable to yogurts having higher amounts of sugar.

The present inventors found that the amount of sucrose or sugar can be reduced below 10% (wt) while maintaining a flavoured yogurt having desired sensory properties. Therefore, the present amount of sucrose or sugar, is preferably 9% (wt) or less, more preferably 8% (wt) or less, even more preferably 7% (wt) or less, more preferably 6% (wt) or less such as 5% (wt) or less, 4% (wt) or less, 3% (wt) or less or even 2% (wt) or less or 1 % (wt) or less. Preferably the present sugar is sucrose.

In a preferred embodiment, the present flavoured yogurt has a texture and / or viscosity which equals the texture and / or viscosity of a similar flavoured yogurt having added sucrose in an amount of more than 5% (wt) or more than 10% (wt). More preferably the present flavoured yogurt has a texture and / or viscosity which texture and / or viscosity equals the texture and / or viscosity of a similar flavoured yogurt having added sucrose in an amount of 5 to 15% (wt), more preferably in an amount of 10 to 15% (wt) or in an amount of 5 to 10% (wt), such as an added sucrose amount of 7% (wt). More preferably, the present flavoured yogurt has an amount of added sucrose of 2% (wt) or less, and a texture and / or viscosity which equals the texture and / or viscosity of a similar flavoured yogurt having an amount of added sucrose of 7% (wt). Most preferably, the present flavoured yogurt has an amount of added sucrose of 2% (wt) or less, and a mouthfeel, such as mouthfeel thickness, which equals the mouthfeel, or mouthfeel thickness, of a similar flavoured yogurt having an amount of added sucrose of 7% (wt). Alternatively, the present flavoured yogurt has a texture and / or viscosity, which equals the mouthfeel, texture and / or viscosity of a similar flavoured yogurt wherein the amount of added sucrose is not reduced.

In a preferred embodiment, the present flavoured yogurt comprises a lactase enzyme (EC 3.2.1.23) or lactase enzyme composition. The presence of a lactase enzyme is advantageous in view of the sweetness of the flavoured yogurt, since lactase may add sweetness by hydrolysing lactose in glucose and galactose. Preferably, the present lactase enzyme or lactase enzyme composition does not has an invertase enzyme activity, or the present lactase enzyme or lactase enzyme composition does not comprise an invertase enzyme (EC 3.2.1 .26). The advantage of using a lactase enzyme without invertase enzyme activity is that the sugar, preferably sucrose, present in the milk is not hydrolysed, which might lead to sweetness loss, texture loss and / or extra syneresis.

Preferably, the present lactase enzyme or lactase enzyme composition does not has an arylsulfatase enzyme activity, or the present lactase enzyme does not comprise an arylsulfatase (EC 3.1.6.1 ). The advantage of using a lactase enzyme without arylsulfatase enzyme activity is the prevention of off flavour formation. This is particularly relevant for flavoured yogurts having a reduced amount of sugar, since off flavours might become more noticeable if the amount of sugar is reduced.

Given the advantageous texture properties of the present flavoured yogurt, in a preferred embodiment, the flavoured yogurt does not comprise a thickening agent, or does not comprise an added thickening agent. The present inventors found that the weakly post acidifying lactic acid bacteria are able to compensate for the loss in texture which is expected as a result of reducing the amount of sugar, and the addition of thickening agents is not needed for generating texture. An added thickening agent means a thickening agent which is not naturally present in the flavoured yogurt. Preferably, the present flavoured yogurt does not comprise a thickening agent selected from the group consisting of carrageenan, xanthan, pectin, alginate, maltodextrin, starch, celluloses, gelatin, locust bean gum and guar. Alternatively, the present flavoured yogurt does not comprise hydrocolloids.

In a preferred embodiment, the present one or more weakly post acidifying lactic acid bacteria comprises one or more lactic acid bacterial strain selected from the group consisting of U, V, W, X, Y and Z. Preferably the present one or more weakly post acidifying lactic acid bacteria comprises U and V, V and W; V and X; V and Y; V and Z; W and X; W and Y; W and Z; X and Y; X and Z; Y and Z; V, W and Z; V, X and Z; V, Y and Z, W, X and Z; W, Y and Z; X, Y and Z; V, W, X and Y or V, W, X, Y and Z. The present inventors found that the perceived mouthfeel of flavoured yogurts prepared with the present lactic acid bacterial strains is comparable to the perceived mouthfeel of flavoured yogurts having higher amounts of sugar. For example the mouthfeel sensory attributes sliminess, melting and astringency are indistinguishable from flavoured yogurts having higher amounts of sugar.

In a preferred embodiment, the present one or more weakly post acidifying lactic acid bacteria comprises a combination of strain U (deposited as CBS139996) and strain V (deposited as CBS140557), a combination of strain V (deposited as CBS140557) and strain W (deposited as CBS140559) or a combination of strain X (deposited as CBS140561 ) and strain Y (deposited as CBS140558).

More preferably, the present one or more weakly post acidifying lactic acid bacteria comprise U, V and Z, V, W and Z or X, Y and Z. The combinations of lactic acid bacterial strains U, V and Z, V, W and Z and X, Y and Z provide a mouthfeel thickness which is comparable to the mouthfeel thickness of flavoured yogurts having higher amounts of sugar. This is unexpected since by reducing the amount of sugar, and thus reducing the amount of dry matter, it was expected that the mouthfeel thickness would reduce as well. Therefore, the combination of strains U, V and Z, V, W and Z or X, Y and Z is suitable to provide a mouthfeel thickness which is comparable with the mouthfeel thickness of a flavoured yogurt having a higher amount of sugar, such as higher than 5% (wt), 6% (wt), 7% (wt), 8% (wt) 9% (wt) or even higher than 10% (wt) sugar.

Further, the present inventors found that the combination of lactic acid bacterial strains U, V and Z, V, W and Z or X, Y and Z is advantageous because reducing the sugar content in the flavoured yogurt has no significant impact on the sour flavour of the flavoured yogurt. This is unexpected since a reduction of sugar generally reveals the acidity of the flavoured yogurt product. Therefore, the combination of strains U, V and Z, V, W and Z or X, Y and Z is suitable to provide a sour flavour which is comparable with the sour flavour of a flavoured yogurt having a higher amount of sugar, such as higher than 5% (wt), 6% (wt), 7% (wt), 8% (wt) 9% (wt) or even higher than 10% (wt) sucrose or sugar.

To meet the sweetness of a full sugar flavoured yogurt, a sweetener is present in the present flavoured yogurt. Examples of sweeteners which may be included in the present flavoured yogurt are steviol glycoside, sucralose, acesulfame K, alitame, aspartame, cyclamate, erythritol, glycyrrhizin, neohesperidin, dihydrochalcone, neotame, saccharin or thaumatin. More preferably a combination of sucralose and acesulfame K.

More preferably, the present sweetener is a steviol glycoside. The steviol glycoside is preferably stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E or rebaudioside M. The steviol glycoside may be a combination of two or more steviol glycosides, for example a combination of at least rebaudioside A and rebaudioside M.

A steviol glycoside for use in the present invention may be one which is derived from a plant source, for example from a plant of the genus Stevia, such as a Stevia rebaudiana plant. Alternatively, a steviol glycoside may be one obtained via an enzymatic process, for example in a process whereby a UDP-glycosyltransferase contacted with a steviol or steviol glycoside substrate. Alternatively, a steviol glycoside may be one which is produced by fermentation of a recombinant host cell which has been modified so that it is capable of production of the steviol glycoside. Suitable recombinant hosts are described in WO2013/110673 and WO2015/007748.

A steviol glycoside suitable for use in the present invention may be provided in the form of a composition comprising the steviol glycoside which is then used in the flavoured yogurt or process of the invention. The steviol glycoside may be stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E or rebaudioside M. A composition comprising the steviol glycoside may comprise at least about 50% by weight of the steviol glycoside, such as at least about 75% by weight of the steviol glycoside, preferably at least about 96% by weight of the steviol glycoside, preferably at least about 97% by weight of the steviol glycoside , such as at least about 98% by weight of the steviol glycoside or more. A composition comprising the steviol glycoside may comprise two or more steviol glycosides.

In a preferred embodiment, the amount of non-sucrose sweetener is within the range of 0.001 % (wt) to 1 % (wt) of the total weight of the flavoured yogurt, preferably within the range of 0.001 % (wt) to 0.1 % (wt). More preferably, the amount of non-sucrose sweetener is within the range of 0.001 % (wt) to 0.05% (wt). The present inventors found that the indicated amounts of non-sucrose sweetener is able to repair the loss in sweetness without introducing off flavours to the flavoured yogurt. For example, a steviol glycoside present in the indicated amounts provides sweetness without introducing a liquorice flavour to the flavoured yogurt. This is unexpected since steviol glycosides may introduce liquorice flavours. Therefore, the present amount of steviol glycoside is preferably suitable to provide sweetness without introducing a liquorice flavour to the flavoured yogurt.

Given the beneficial sensory profile of the present flavoured yogurt, the present flavoured yogurt has an increased mouthfeel thickness, an increased creamy mouthfeel, and/or an improved sour flavour if compared with a flavoured yogurt which has not been fermented with the composition comprising the present weakly post acidifying lactic acid bacteria. Alternatively, the present flavoured yogurt has an increased mouthfeel thickness, an increased creamy mouthfeel, and/or an improved sour flavour if compared with a flavoured yogurt wherein no sweetener and / or lactase enzyme has been added.

In a preferred embodiment, the present flavoured yogurt has a pH profile during storage at 4°C between 1 and 40 days after fermentation, preferably at 4°C between 20 and 40 days after fermentation, which does not fluctuate more than 0.3 unit. Preferably the pH does not fluctuate more than 0.2 pH, more preferably said pH does not fluctuate more than 0.1 pH.

According to another aspect, the present invention relates to a process for the production of a flavoured yogurt, or a yogurt, comprising fermenting milk having an amount of sucrose, added sucrose or sugar, of 10% (wt) or less with a weakly post acidifying starter culture to provide the flavoured yogurt, or yogurt, and comprising adding a non-sucrose sweetener to the milk having an amount of sucrose, added sucrose or sugar 10% (wt) or less or adding a non-sucrose sweetener to the provided flavoured yogurt, or yogurt. It has been found that the present process is able to compensate for the loss in texture resulting from reducing the amount of added sugar, or sucrose, by using a weakly post acidifying starter culture. The starter cultures of the invention have the ability to provide a yogurt having desired properties, while reducing the amount of added sugar or sucrose. More specifically, the cultures of the invention have the ability to provide a yogurt having a reduced amount of added sugar or sucrose, while maintaining the texture.

The non-sucrose sweetener is added to the present milk having an amount of sucrose, added sucrose or sugar 10% (wt) or less or to the provided flavoured yogurt directly. If added to the milk the non-sucrose sweetener is present during fermentation. If added to the provided flavoured yogurt, the non-sucrose sweetener may be added to the provided flavoured yogurt directly or to a fruit preparation prior to its mixture with the provided flavoured yogurt. The non- sucrose sweetener may also be added during mixing of a fruit preparation with the provided flavoured yogurt.

The present milk having an amount sucrose or sugar of 10% (wt) or less has preferably having an amount of sucrose or sugar of 9% (wt) or less, more preferably 8% (wt) or less, even more preferably 7% (wt) or less, more preferably 6% (wt) or less such as 5% (wt) or less, 4% (wt) or less, 3% (wt) or less or even 2% (wt) or less or 1 % (wt) or less.

Preferably, the present process for the production of a flavoured yogurt is a process for the production of a flavoured yogurt having an amount of sucrose or sugar of 10% (wt) or less, preferably an amount of sucrose or sugar of 9% (wt) or less, more preferably 8% (wt) or less, even more preferably 7% (wt) or less, more preferably 6% (wt) or less such as 5% (wt) or less, 4% (wt) or less, 3% (wt) or less or even 2% (wt) or less or 1 % (wt) or less.

The present step of fermenting milk having an amount of sugar or sucrose of 10% (wt) or less with a composition comprising a weakly post acidifying starter culture to provide the flavoured yogurt preferably comprises maintaining the milk at a fermentation temperature until the desired pH value in the range of 4.0 and 5.0, preferably a pH value in the range of 4.0 to 4.7 and more preferably in the range of 4.2 to 4.5 is reached (end of fermentation). The pH is preferably in the range of 4.2 - 5.0. The length of the fermentation period is not critical for the present invention and can be determined easily by the skilled person. The fermentation temperature may be in the range of 30°C-45°C, more preferably in the range of 37°C-42°C.

In a preferred embodiment, the present process further comprises a step of cooling the milk when a pH value in the range of 4.0 and 5.0 is reached and / or a step of packaging the provided flavoured yogurt. Preferably, the provided flavoured yogurt is packaged in a package having a volume of more than 50 ml, such as from 50 ml to 1 liter.

In a preferred embodiment, the present process further comprises adding a lactase enzyme (EC 3.2.1 .23) composition to the milk having an amount of sugar or sucrose of less than 10% (wt) or to the provided flavoured yogurt. Preferably, the lactase enzyme is added to the present milk before or together with the present starter culture. Alternatively, the lactase enzyme is added during fermentation. Advantageously, lactose hydrolysis by the lactase enzyme and fermentation with the starter culture is carried out simultaneously. The advantage of adding a lactase enzyme is the hydrolysis of lactose into glucose and galactose, both of which provide a stronger sweetness than lactose.

In a further preferred embodiment, the present weakly post acidifying starter culture comprises one or more lactic acid bacterial strain selected from the group consisting of U, V, W, X, Y and Z. Preferably, the present weakly post acidifying culture comprises U and V, V and W; V and X; V and Y; V and Z; W and X; W and Y; W and Z; X and Y; X and Z; Y and Z; U, V and Z; V, W and Z; V, X and Z; V, Y and Z, W, X and Z; W, Y and Z; X, Y and Z; V, W, X and Y or V, W, X, Y and Z. The combination of U, V and Z; V, W and Z; or X, Y and Z are highly preferred. More preferably, the present weakly post acidifying culture comprises the combination of strain U (deposited as CBS139996) and strain V (deposited as CBS140557), a combination of strain V (deposited as CBS140557) and strain W (deposited as CBS140559) or a combination of strain X (deposited as CBS140561 ) and strain Y (deposited as CBS140558). In yet another preferred embodiment, the present sweetener is a steviol glycoside. The steviol glycoside is preferably stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E or rebaudioside M. The steviol glycoside may be a combination of two or more steviol glycosides, for example a combination of at least rebaudioside A and rebaudioside M.

In a preferred embodiment, the present provided flavoured yogurt has an increased mouthfeel thickness, if compared with a flavoured yogurt which has not been fermented with the weakly post acidifying starter culture. Preferably, the present provided flavoured yogurt has a mouthfeel thickness which is comparable with a similar yogurt having an amount of sugar of 7% (wt).

In another preferred embodiment, the present provided flavoured yogurt has an increased creamy mouthfeel if compared with a flavoured yogurt which has not been fermented with the weakly post acidifying starter culture. Preferably, the present provided flavoured yogurt has a creamy mouthfeel which is comparable with a similar yogurt having an amount of sugar of 7% (wt).

In yet another preferred embodiment, the present provided flavoured yogurt has an improved, or decreased, sour flavour if compared with a flavoured yogurt which has not been fermented with the weakly post acidifying starter culture. Preferably, the present flavoured yogurt has a sour flavour which is comparable with a similar yogurt having an amount of sugar of 7% (wt).

In a further preferred embodiment, the present provided flavoured yogurt has a less, or decreased, liquorice flavour, or, liquorice root flavour, if compared with a flavoured yogurt which has not been fermented with the weakly post acidifying starter culture. Preferably, the present flavoured yogurt has a liquorice flavour which is comparable with a similar yogurt having an amount of sugar of 7% (wt).

In yet another preferred embodiment, the present provided flavoured yogurt, or provided flavoured yogurt, has a shear stress, and / or viscosity, which is lower than the shear stress and / or viscosity of a similar flavour yogurt having an higher amount of sugar. Preferably, the shear stress and / or viscosity is lower than the shear stress and / or viscosity of a yogurt having an amount of sugar of 7% (wt).

Given the advantageous flavour and texturizing properties of the present weakly post acidifying starter culture, the present invention relates, according to another aspect, to the use of a lactic acid bacteria chosen from U, V, W, X, Y and Z, or chosen from the combination of strain U (deposited as CBS139996) and strain V (deposited as CBS140557), a combination of strain V (deposited as CBS140557) and strain W (deposited as CBS140559) or a combination of strain X (deposited as CBS140561 ) and strain Y (deposited as CBS140558) for reducing the amount of added sugar, such as sucrose, in yogurt, preferably in a flavoured yogurt, preferably for reducing the amount of added sugar, such as sucrose, with at least 30%, more preferably at least 50%, most preferably at least 70%. Preferably, for reducing the amount of sugar in a yogurt which does not comprise a thickening agent. The present inventors found that the present lactic acid bacterial strains are able to compensate the loss in texture, mouthfeel thickness, creamy mouthfeel and for the increase in sour flavour in a yogurt wherein the amount of sugar has been reduced. This is unexpected in view of the flavour and texturizing properties of sugar, particularly if a sugar reduction of more than 30% is reached. Therefore, the present invention relates to the use of the present lactic acid bacteria, preferably chosen from the combination of strain U (deposited as CBS139996) and strain V (deposited as CBS140557), a combination of strain V (deposited as CBS140557) and strain W (deposited as CBS140559) or a combination of strain X (deposited as CBS140561 ) and strain Y (deposited as CBS140558) for providing thickness, preferably mouthfeel thickness in a flavoured yogurt product.

Preferably the present lactic acid bacterial strain comprises U and V, V and W; V and X; V and Y; V and Z; W and X; W and Y; W and Z; X and Y; X and Z; Y and Z; U, V an Z; V, W and Z; V, X and Z; V, Y and Z, W, X and Z; W, Y and Z; X, Y and Z; V, W, X and Y or V, W, X, Y and Z. The combination of U, V and Z; V, W and Z; or X, Y and Z is highly preferred.

In a preferred embodiment, the present invention relates to the use of the present lactic acid bacteria for providing texture, mouthfeel thickness and / or a creamy mouthfeel in a flavoured yogurt having an amount of sugar, or sucrose, of 10% (wt) or less, preferably an amount of sugar, or sucrose, of 9% (wt) or less, more preferably 8% (wt) or less, even more preferably 7% (wt) or less, more preferably 6% (wt) or less such as 5% (wt) or less, 4% (wt) or less, 3% (wt) or less or even 2% (wt) or less or 1 % (wt) or less.

The invention is further elucidated in the illustrative example below, wherein reference is made to figure 1. Figure 1 is a spider plot with the key sensory attributes for sweetened yoghurt of UVZ in recipes containing 7% sucrose (black line) and 2% sucrose + steviol glycoside (grey line). Chemical after taste and shininess were the only two attributes out of 29 assessed that showed a statistically significant, but small, difference between the two samples.

MATERIALS AND METHODS 1. Lactic acid bacteria cultures.

Table 1. lactic acid bacterial strains

Strain CBS number bacteria

U CBS 139996 Streptococcus thermophilus

V CBS 140557 Streptococcus thermophilus

W CBS 140559 Streptococcus thermophilus

X CBS 140561 Streptococcus thermophilus

Y CBS 140558 Streptococcus thermophilus Strain CBS number bacteria

Z CBS 140562 Lactobacillus delbrueckii ssp. bulgaricus

All cultures were deposited at the Centraalbureau voor Schimmelcultures (Fungal Biodiversity Centre), Uppsalalaan 8, 3584 CT Utrecht, The Netherlands under the provisions of the Budapest Treaty, on 6 May 2015: CBS139996 and on 29 September 2015: CBS140557, CBS140558, CBS140559, CBS140561 , CBS140562.

Table 2. lactic acid bacteria cultures

2. Lactase

The commercial product Maxilact®LGI 5000 (obtainable from DSM, The Netherlands containing 5000 NLU/g), was added to the milk alongside the starter culture at a rate of 5000 NLU per liter milk.

3. Non-sucrose sweeteners

Rebaudioside A having a purity of 95% was added to the milk with a concentration of 0.1 % gram / liter milk, alongside the starter culture and lactase. The rebaudioside A was fermentative rebaudioside A obtained as disclosed in WO2013/1 10673. Alternatively, an acesulfame K sucralose blend (30 wt% acesulfame K and 70 wt% sucralose) was added to the milk with a concentration of 0.0062% w/w). 4. Yogurt preparation

The milk used is obtained by supplementing pasteurized skimmed milk (Campina, The Netherlands) with skimmed milk powder and sucrose (to 2 and 7%). If steviol glycosides were added, they were added to the milk at this stage as well. The final recipe is described in the different examples. The milk mixture is pasteurized at 85°C for 30 minutes. The pasteurized milk is cooled back to the fermentation temperature (38 or 42°C) and inoculated with the culture to be tested at a rate of 0.02% (w/w) or 4 units per 1000 liters of milk. If lactase was added, it was added to the milk together with the culture. Once a pH of 4.60 is reached, the yogurt is smoothened by pumping the yogurt through sieves (poresize 750μηη and 350μηη) whilst being cooled to 20°C. The yogurt is then filled out into suitable containers. The yogurt cups are then stored at 4°C until sensory analysis.

5. Yogurt recipes

The following recipes were used in the Examples. All additions are weight percentages of the total milk recipe.

Table 3. Yogurt recipes

"total of the yogurt recipe comprising semi skimmed milk, skimmed milk powder and sucrose.

6. Sensory analysis

Descriptive sensory analysis was done by using the Quantitative Descriptive Analysis Method (Stone, H. and Sidel, J.L. "Sensory Evaluation Practises" 3^rd edition, 2004). First, the panellists developed a list of attributes including definitions by means of evaluating a wide variety of references and a wide array of yogurts. Secondly, training sessions were organized to enable panellists to learn to consistently differentiate and replicate the yogurt samples. During the actual QDA measurements the intensities of the selected attributes were obtained per product by the FIZZ (Biosystems; France) sensory data acquisition system, using unstructured line scales ranging from 0 - 100. The products were offered semi-monadically and evaluated twice by the panellists (n=14) by means of a Balanced Complete Block design to avoid sequence effects. Statistical analysis of the data was done by analysis of variance with Fisher's least significant difference (LSD) as a post hoc test (SenPaq) and modelled using Principle Component analysis (PCA) (SenPaq).

7. Attributes of stirred yogurt

For the appearance (i.e. the visual assessment of surface of the yogurt) the following attributes may be determined:

• Shininess which is the degree in which the surface of the yogurt reflects the light.

· Whiteness which is the whiteness of the surface of the yogurt. For the structure (i.e. the visual assessment of texture by means of spoon) the following attributes may be determined after stirring the yogurt 5 times before assessing the structure.

• Ropiness which is the degree in which the yogurt runs from the spoon. The anchors may range from short to long. Short means that there is no formation of threads. Long means that the product forms almost one long thread when it runs from the spoon.

• Thickness which is the force needed for stirring the yogurt.

• Smoothness which is the degree in which the product is smooth without grittiness, grains and lumps, assessed at the back of the spoon.

For the odour the following attributes may be determined

• Intensity which is the general and overall odour intensity.

• Sour which is the intensity of the sour odour.

• Sweet which is the intensity of the sweet odour.

• Fruity which is the degree in which a fruity odour is present in the product (e.g. apple, lemon, lime).

• Off-odour which is a group of odours which can't be assessed by means of other odour attributes such as manure, stable, cauliflower, sulphur and medicinal odours.

For the mouthfeel (assessment of mouthfeel, when the product is inside the mouth) the following attributes may be determined

• Thickness which is the degree in which the product feels thick in the mouth. This sensation can be best perceived between tongue and palate.

• Grainy which is the degree in which the product feels grainy/powdery in the mouth.

• Creaminess which is the full and creamy feeling in the mouth. The product does not feel rough; it does not feel dry and gives a velvety coating in the mouth.

• Slimy which is the degree in which the product feels slippery and forms threads in the mouth.

This sensation has to be assessed by pushing the product against the palate with the tongue.

• Melting which is the degree (slow - quick) in which the product mixes with saliva and disappears out of the mouth.

• Astringent which is the degree in which an astringent feeling is present in your mouth after you swallowed the product (the sensation can be mimicked with red wine and the "skin" of a nut).

For the flavour the following attributes may be determined

• Intensity which is the general and overall flavour intensity.

• Sour which is the degree in which the product tastes sour (mimicked by citric acid). • Milky/creamy which is the degree in with a milk/cream flavour is perceived (this can be mimicked with full fat milk or cream).

• Green which is the degree in which a green flavour is perceived. It can be best described as the green flavour which can be perceived when eating a green apple. This can be mimicked with acetaldehyde.

• Sweet which is the degree in which the product has a sweet taste.

• Bitter which is the degree in which the product has a bitter taste.

For the after taste (assessment after swallowing the product) the following attributes may be determined

• Intensity which is the general and overall aftertaste intensity.

• UHT which is the degree in which the aftertaste of sterilized milk (long life milk) is present (e.g. caramel or cooked milk but not burnt milk).

• Chemical which is the intensity of a possible chemical aftertaste such as vitamins, rubber and artificial sweeteners.

• Length which is the duration (in time) of the aftertaste.

For the after feel (assessment after swallowing the product) the following attributes may be determined

• Astringent which is the degree in which an astringent feeling is present in the mouth after the product has been swallowed (the sensation can be mimicked with red wine and the "skin" of a nut).

• Pungent which is the degree in which a pungent feeling remains after swallowing the yogurt.

• Fat coating which is the degree in which a fat coating remains in the mouth after the products is swallowed (filming effect).

Example 1

Sensory effect of lactic acid bacterial culture on sucrose reduction.

Yogurt was made according to the method described in the Materials and Methods with starter cultures VWZ and XYZ as indicated in table 2 and the recipe A and B as defined in Table 3. Four different recipe A yogurts were prepared: two recipe A yogurts with the starter culture VWZ or XYZ; and two recipe A yogurts with the culture VWZ or XYZ supplemented with lactase enzyme and steviol glycoside (as depicted as ++ in table 4 below). Recipe B was fermented with starter culture VWZ or XYZ, without the addition of lactase enzyme and steviol glycoside. Subsequently, on day 7 after fermentation, all yogurts were subjected to a sensory analysis including the attributes as described in Materials and Methods. The results of the sensory analysis are shown in table 4 below.

Table 4

++ determines the presence o lactase and steviol glycoside during ermentation The above results show that both starter cultures of the invention are able to provide a flavoured yogurt having an amount of sucrose of 2% (wt), wherein the structural thickness, the mouthfeel thickness, the mouthfeel attributes slimy, melting and astringent are comparable to 7% sucrose flavoured yogurt. Furthermore, by addition of steviol glycoside and lactase enzyme, the sweet flavour is repaired to the sweet flavour of the 7% sucrose flavoured yogurt, wherein the liquorice flavour has not been increased significantly.

Additionally, strain XYZ in combination with lactase enzyme and steviol glycoside is able to repair the loss in creamy mouthfeel and the increase in sour flavour which is the result of the reduction of sucrose from 7% (wt) to 2% (wt).

Example 2

Sensory effect of lactic acid bacterial culture on sucrose reduction. Yogurt was made according to the method described in the Materials and Methods with starter culture UVZ as indicated in table 2 and the recipe A and B as defined in Table 3. Subsequently, the yogurts were subjected to a sensory analysis including the attributes as described in the Materials and Methods. The results of the sensory analysis, performed on day 7 after fermentation, are shown in figure 1. Figure 1 shows that the combination of UVZ and steviol glycoside is able to provide a flavoured yogurt having an amount of sucrose of 2% (wt), wherein the key texture and flavour attributes are comparable to a 7% sucrose flavoured yogurt.

Example 3 Sensory effect of various alternative sweeteners on sucrose reduction.

To test the effect of the present invention with various sweeteners, example 1 has been repeated in combination with alternative sweeteners. As sweeteners a fermentative steviol glycoside or a acesulfame K sucralose blend were used according to the materials & methods. Subsequently, on day 7 after fermentation, the yogurts were subjected to a sensory analysis including the attributes as described in Materials and Methods. The results of the sensory analysis are shown in table 5 below.

Table 5

A = Acesul ame-K ucralose blend; = ermentative steviol glycoside; L =

The above results show that the combination of XYZ, fermentative steviol glycoside and lactase as well as the combination of XYZ, acesulfame-K/sucralose and lactase is able to provide a flavoured yogurt having an amount of sucrose of 2% (wt), wherein the sweet flavour is the same as a 7% sucrose flavoured yogurt. Moreover, the impact on other sensory attributes is marginal. Applicant's or agent's file reference number 31474-WO-PCT | International application No

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Rodriguez, Ana

INDICATIONS RELATING TO A DEPOSITED MICROORGANISM

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INDICATIONS RELATING TO A DEPOSITED MICROORGANISM

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Rodriguez, Ana

INDICATIONS RELATING TO A DEPOSITED MICROORGANISM

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Rodriguez, Ana

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Form PCT/RO/134 (My 1992)

Claims

1. Flavoured yogurt comprising added sucrose in an amount of 10% (wt) or less and comprising one or more weakly post acidifying lactic acid bacteria and a non-sucrose sweetener.

2. Flavoured yogurt according to claim 1 , having an amount of added sucrose of 5% (wt) or less.

3. Flavoured yogurt according to claim 1 or claim 2, further comprising a lactase enzyme (EC 3.2.1.23).

4. Flavoured yogurt according to any of the preceding claims, which does not comprise a thickening agent.

5. Flavoured yogurt according to any of the preceding claims, wherein the one or more weakly post acidifying lactic acid bacteria comprises a combination of strain U (deposited as CBS139996) and strain V (deposited as CBS140557), a combination of strain V (deposited as CBS140557) and strain W (deposited as CBS140559) or a combination of strain X (deposited as CBS140561 ) and strain Y (deposited as CBS140558).

6. Flavoured yogurt according to any of the preceding claims, wherein the non- sucrose sweetener is a steviol glycoside, preferably one or more steviol glycoside selected from rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E and rebaudioside M.

7. Flavoured yogurt according to any of the preceding claims, wherein the amount of non-sucrose sweetener is within the range of 0.001 % (wt) to 1 % (wt), preferably within the range of 0.001 % (wt) to 0.1 % (wt).

8. A process for the production of a flavoured yogurt, comprising fermenting milk having an amount of sucrose of 10% (wt) or less with a weakly post acidifying starter culture to provide the flavoured yogurt, and comprising adding a non-sucrose sweetener to the milk having an amount of sucrose 10% (wt) or less or to the provided flavoured yogurt.

9. Process according to claim 8, further comprising adding a lactase enzyme (EC

3.2.1.23) composition to the milk having an amount of sucrose of less than 10% (wt) or to the provided flavoured yogurt.

10. Process according to claim 8 or claim 9, comprising fermenting milk having an amount of sucrose of 5% (wt) or less.

1 1. Process according to any of the preceding claims, wherein the weakly post acidifying starter culture comprises a combination of strain U (deposited as CBS139996) and strain V (deposited as CBS140557), a combination of strain V (deposited as CBS140557) and strain W (deposited as CBS140559) or a combination of strain X (deposited as CBS140561 ) and strain Y (deposited as CBS140558).

12. Process according to any of the preceding claims, wherein the non sucrose sweetener is a steviol glycoside, preferably one or more steviol glycoside selected from rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E and rebaudioside M.

13. Process according to any of the preceding claims, wherein the provided flavoured yogurt has an increased mouthfeel thickness, an increased creamy mouthfeel, and/or an improved sour flavour if compared with a flavoured yogurt which has not been fermented with the weakly post acidifying starter culture.

14. Process according to any of the preceding claims, wherein the provided flavoured yogurt has a decreased liquorice flavour if compared with a flavoured yogurt which has not been fermented with the weakly post acidifying starter culture.

15. Use of a lactic acid bacteria chosen from the combination of strain U (deposited as CBS139996) and strain V (deposited as CBS140557), a combination of strain V (deposited as

CBS140557) and strain W (deposited as CBS140559) or a combination of strain X (deposited as CBS140561 ) and strain Y (deposited as CBS140558) for reducing the amount of added sugar in yogurt, preferably in flavoured yogurt, preferably for reducing the amount of added sugar with at least 30%, more preferably at least 50%, most preferably at least 70%.

16. Use of a lactic acid bacteria chosen from the combination of strain U (deposited as CBS139996) and strain V (deposited as CBS140557), a combination of strain V (deposited as CBS140557) and strain W (deposited as CBS140559) or a combination of strain X (deposited as CBS140561 ) and strain Y (deposited as CBS140558) for providing texture, mouthfeel thickness and / or a creamy mouthfeel in a flavoured yogurt having an amount of sucrose of 10% (wt) or less.