TASTE AND FLAVOUR MODULATION BY BIOTRANSFORMATION IN MILK

PRODUCTS

Field of the invention

The present invention relates to a method for promoting natural tastes and flavours in milk-based products, to a method for manufacturing said milk products and to the products thus obtainable.

Background of the invention

Traditionally, in the production of milk products such as cheese, butter etc., the characterising flavours are developed during fermentation of the milk source with bacteria.

In recent years, novel flavouring agents which can be added to milk products for taste improvement have been developed. These flavouring agents are traditionally obtained by treating a milk source with enzymes, microorganisms etc. For instance, a number of publications describe fermentation of a milk source with microorganisms and enzymes such as lipases, proteases etc. to produce aromas which can be isolated and used in the manufacture of flavoured milk products such as cheese, milk, fermented milk etc. Amongst these publications are CA 1220075, EP 137536, JP 2003-250482, JP 3127962, JP 5049385, JP 2002-142713, JP 5091851, JP 58043755, JP 6125733, JP 64002549, EP 1053689, EP 1186244, US 3858492, JP 74015785, JP 2004-267126, JP 6319448, US 6635303 Bl, JP 80043742, JP 2005-151895, JP 30120942, JP 74015784, JP 3175978, JP 2004-236638, JP 4169166, JP 9037735, US 3975544, WO 0147366. The flavours thus obtained are

described as "dairy" flavours and therefore act as flavour enhancers to dairy products .

However, there are limited reports on using a process similar to that described above in order to provide flavours distinct from "dairy" flavours such as, for example, chocolate, fruity flavours etc.

WO 02/00845 describes a way to obtain chocolate-flavoured fermented products by fermenting a milk source with a mixed micro-organism culture. This method is however of limited scope in terms of the flavours obtained and in terms of the applications.

In order to achieve a variety of different, distinct flavours, artificial synthetic flavourings are normally added to the milk products. The use of artificial flavourings is quite often linked with a negative impression by the consumers .

There is thus a need to provide a wide variety of flavours in a natural way which can be used in a wide range of foods, and which thus avoids the use of artificial flavourings .

Summary

Accordingly, the present object is achieved by means of the features of the independent claims. The dependent claims develop further the central idea of the invention.

The present invention proposes in a first aspect a method to promote a non-savoury flavour in a food product, comprising the steps of:

a) adding at least one amino acid to a milk source or adding a protease and/or peptidase to the milk source in order to obtain at least one amino acid from a milk protein, b) adding at least one micro-organism to the milk source, wherein the micro-organism is selected for producing at least one enzyme capable of converting the amino acid, c) fermenting the milk source d) adding a lipase and/or lactase prior to and/or during and/or after fermentation e) optionally drying the fermented milk source and f) adding said fermented milk source to the food product .

In a second aspect, the invention provides a method for flavouring milk powder comprising the steps of: a) adding at least one amino acid to a milk source or adding a protease and/or peptidase to the milk source in order to obtain at least one amino acid from a milk protein, b) adding at least one micro-organism to the milk source, wherein the micro-organism is selected for producing at least one enzyme capable of converting the amino acid, c) fermenting the milk source d) optionally drying the fermented milk source and e) adding said fermented milk source to the milk powder or milk powder precursor.

A method for flavouring a frozen dessert comprising the steps of :

a. adding at least one amino acid to a milk source or adding a protease and/or peptidase to the milk source in order to obtain at least one amino acid from a milk protein, b. adding at least one micro-organism to the milk source, wherein the micro-organism is selected for producing at least one enzyme capable of converting the amino acid, c. fermenting the milk source d. optionally drying the fermented milk source and e. adding said fermented milk source to the frozen dessert mix also falls under an aspect of the present invention.

An alternative method for flavouring a food product comprising the steps of : a) adding at least one amino acid to a milk source or adding a protease and/or peptidase to the milk source in order to obtain at least one amino acid from a milk protein b) optionally adding L-rhamnose to the milk source c) adding a lipase and/or lactase prior to and/or during and/or after enzymatic treatment d) heat treatment the milk source e) optionally drying the treated milk source and f) adding said treated milk source to the food product is also part of the present invention.

Further, a frozen or chilled dessert obtainable by a method according to any of claims 1 or 7 to 23 and a milk powder obtainable by a method according to any of claims 2,3 or 6 to 23 is provided.

A liquid flavour concentrate obtainable by a) adding at least one amino acid to a milk source or adding a protease and/or peptidase to the milk source in order to obtain at least one amino acid from a milk protein, b) adding at least one micro-organism to the milk source, wherein the micro-organism is selected for producing at least one enzyme capable of converting the amino acid and c) fermenting the milk source falls under an aspect of the present invention.

A milk powder-like product comprising per kg of product 1- 20000mg, preferably 10-500mg butanoic acid, 10-lOOOOmg, preferably 100-3000mg hexanoic acid, 10-6000mg, preferably 100-3000mg octanoic acid, 2-13000mg, preferably 50-1000mg decanoic acid also forms part of the present invention.

The invention relates to the use of a milk powder-like product according to claims 28 to 33 in the manufacture of ice cream, milk powder, chocolate-based product, dairy product, creamer, cocoa beverage, culinary product, infant formula or health care, product .

Finally, a method for flavouring a food product comprising the steps of : a) adding at least one amino acid to a milk source or adding a protease and/or peptidase to the milk source in order to obtain at least one amino acid from a milk protein b) optionally adding L-rhamnose to the milk source c) adding a lipase and/or lactase prior to and/or during and/or after enzymatic treatment

d) heat treatment the milk source e) optionally drying the treated milk source and f) adding said treated milk source to the food product also forms part of the invention.

Figures

The present invention is further described hereinafter with reference to some of its embodiments shown in the accompanying drawings in which:

Fig. 1 is a flow-chart showing method steps in the production of a sweet honey or sweet honey and cream butter flavoured milk powder Fig. 2 is a flow-chart showing method steps in the production of a malt and chocolate or a malt, chocolate and honey flavoured milk powder

Fig. 3 depicts a series of chemical reactions which may lead to compounds responsible for the honey flavour.

Fig. 4 depicts a series of chemical reactions which may lead to compounds responsible for a cocoa and malt flavour. Fig. 5 depicts a chemical reaction which may lead to compounds responsible for a cocoa/coffee flavour. - Fig. 6 is a flow-chart showing method steps in the production of a liquid flavouring product .

Detailed description of the invention

The present invention is concerned with the production of natural flavours which can be used in a number of food applications.

Thus, the invention relates, in a first aspect, to a method to promote a non-savoury flavour in a food product. By "non-savoury" flavour is meant a flavour which is not savoury, i.e. not salty, spicy, sharp etc. A typical savoury flavour would be a cheesy, meaty, salty flavour. However, the invention does not relate to such flavours but to "non-savoury" flavours. The flavours of the present invention thus include "sweet", creamy etc. flavours. Additionally, the flavours of the invention preferably do not include yogurt flavours.

In a preferred embodiment, the flavour is selected from honey, caramel, cocoa, malt, cream-caramel, sweet-honey cream, cocoa-cream, coffee, cream, butter, vanilla, nutty, chocolate and any combination or sub-combination thereof.

Such method for promoting said non-savoury flavour in a food product involves, in a first step, the addition of at least one amino acid to a milk source. Alternatively, the addition of a protease and/or peptidase to a milk source in order to obtain at least one amino acid from a milk protein is possible.

In the present invention, the enzymes may be provided as such, in the form of enzyme preparations, in the form of immobilised enzymes and may even be provided by appropriate micro-organisms.

Any amino acid may be used. Preferably, the amino acid is selected from phenylalanine, leucine, isoleucine, valine and mixtures thereof .

The amino acid may be added to the milk source in an amount of 0.01 to 5 wt%, preferably 0.01-2 wt%, more preferably in an amount of 0.03-1.0 wt%, most preferably 0.05-0.3 wt% on dry matter.

In a preferred embodiment, a source of alpha-keto acid, such as alpha-ketoglutarate, alpha-ketoisocaproate, alpha- ketoisovalerate, similar amino acceptors or mixtures thereof may also be added to the milk source. Preferably, alpha-ketoglutarate is used. These naturally occurring compounds help contribute to the enzymatic pathway of the amino acid, as occurs in the subsequent steps of the present method.

Optionally, cofactors for improving fermentation efficiency such as manganese or magnesium salts may also be added to the milk source.

The milk source may be UHT-treated, pasteurised or non- pasteurised. It may be selected from milk (full fat, skimmed or semi-skimmed) , fresh milk, recombined milk, cream, buttermilk, whey, milk containing vegetable fat etc. and any mixtures thereof. Any type of milk can be used, such as cow, sheep, goat or buffalo milk or any mixtures thereof .

If the milk source is non-pasteurised, the milk source and amino acid mixture may be pasteurised or ultra-high

temperature treated or sterilised under typical conditions in the range of 70 ⁰ C to 150 ⁰ C over 2s to 20 min, e.g. 125 ⁰ C for 20 s. Alternatively, the milk source may be heat-treated first prior to amino acid addition, or prior to treatment with a protease and/or peptidase.

Optionally, L-rhamnose may be added to the milk source.

To the milk source is then added, in a second step, at least one micro-organism, wherein the micro-organism is selected for producing at least one enzyme capable of modifying the amino acid.

In the present method, the micro-organism is selected in particular for its ability to produce an enzyme, the action of which, depending on the substrates, produces direct or indirect conversion products which are responsible for a particular flavour.

Such enzymes are typically transamidase, decarboxylase, dehydrogenase enzymes. Other enzymes such as aldolase may also be produced. Optionally, additives such as protease and/or peptidase may be added together with the microorganisms .

The micro-organisms used in the present method may be selected from the group consisting of Betacoccus, Lactobacillus, Propionibacteria, Streptococcus, Bifidobacterium, Penicillium, Brevibacterium, Arthrobacter, Corynebacterium, Saccharomyces, Debaromyces, Lactococcus and any mixtures thereof and/or mixtures of different strains of the same micro-organism. Preferably, it is a Lactococcus lactis and more preferably it is a

Lactococcus lactis including subspecies. Most preferably, it is a Lactococcus lactis subsp. lactis and/or Lactococcus lactis subsp. lactis biovar. diacetylactis. It may be added in the form of a living microbial starter culture or in the form of a microbial culture after cell lysis or a mixture of both.

Strains of Lactococcus lactis which are commercially available and which are suitable for use in the present invention are for example Lactococcus lactis ATCC 29146, Lactococcus lactis subsp. lactis DSM 4366, or still mesophilic aromatic culture (mix) , type LD, F-DVS XT-313- eXact, Chr. Hansen.

Under fermentation conditions, the micro-organism produces at least some enzymes capable of converting the amino acid into further components. Said micro-organisms are capable of producing enzymes such as transamidase and/or decarboxylase and/or dehydrogenase enzymes, which will act on the amino acid(s) present in the fermentation broth. Aldolase may also be produced by the micro-organism.

Thus, in an alternative embodiment, the micro-organism may be replaced or supplemented with enzymes or enzyme mixes which act on the amino acid to produce the desired flavour character. Furthermore, cell lysates can also additionally be used to modify the amino acid.

Additional enzymes, such as lipase, lactase etc. may also be added to the fermentation broth at the start of the fermentation or at a later stage. These may be added in the form of enzyme preparations or micro-organisms which produce said enzymes. Thus, a lipase and/or lactase may be

added prior to and/or during and/or after fermentation. This is advantageous for further modulation of the flavour obtained.

The mix is then fermented. Under fermentation conditions, the released enzymes will react with the amino acid(s) present in the fermentation mixture to provide direct or indirect conversion products which are responsible for particular aromas. These are, for instance, from the group of 3 -methyl-butanal, 2-methyl-butanal, 2-methyl-1- propanal, 2-phenylacetaldehyde, 2 , 3-butandione (diacetyl) , 3-hydroxy-2-butaneone (acetoin) , 2-phenyl-2-butenal and other phenyl-aldehydes (ClO-16) , benzaldehyde, 5-methyl-2- phenyl-cis-2-hexenal (cocal) , 2 , 3 , 5 , 6-tetramethyl pyrazine, 2 , 4 , 5-trimethyl oxazole, delta-octalactone (5- octanolide) , delta-decalactone (5-decanolide) , delta- dodecalactone (5-dodecanolide) , butanoic acid, hexanoic acid, octanoic acid, decanoic acid, dodecanoic acid, 4- hydroxy-2, 5-dimethyl-3 (2H) -furanone (furaneol) . By phenyl- aldehydes (ClO-16) is meant phenyl-aldehydes having a carbon chain of 10 to 16 carbon atoms.

In this context, figures 3, 4 and 5 depict reaction pathways which may lead to some compounds responsible for honey flavour, for cocoa and malt flavour, or for cocoa/coffee flavour respectively. It is clear to the skilled person that figures 3 to 5 are not illustrative of the full process nor of all the reactions occurring when carrying out the methods of the invention. It is also clear that other reactions may occur during fermentation and that other compounds may be produced which provide further modulation of the flavours.

In order to modulate the flavour of the fermentation broth, the amino acid used may be chosen accordingly.

For instance, it has been found that when the amino acid selected is phenylalanine, the direct or indirect conversion products when subjected to a transamidase and/or decarboxylase and/or dehydrogenase enzyme are responsible for a honey-like flavour.

When the amino acid is selected from L-leucine, L- isoleucine, L-valine or mixtures thereof, the particular flavours obtained are those of malt and cocoa.

When rhamnose is optionally used in the starting milk basis, and the mixture before or after fermentation is heated for about 90 minutes at about 90 ⁰ C, the resulting aroma is that of caramel.

Thus flavours such as honey, caramel, cocoa, malt, cream- caramel, sweet-honey cream, cocoa-cream, coffee, cream, butter, vanilla, nutty, chocolate etc. and any combination or sub-combination thereof may also be obtained by the method of the invention. Furthermore, the tastes obtained by the method of the invention may be creamy, mouth- coating, long-lasting tastes.

Typically, the fermentation is allowed to take place for about between 2 and 24 hours, preferably between 3 and 12 hours, more preferably about 6 hours at a temperature between 8 and 50 ⁰ C. Preferably, the fermentation is carried out for about 6 hours at about 30 ⁰ C.

After a period of time sufficient to produce the desired compounds, the fermentation broth may be cooled to produce a liquid flavour concentrate. A liquid flavour concentrate obtainable by the steps described above is part of the present invention. Alternatively, it may be dried, preferably spray-dried, in order to produce a milk powder- like product having a modulated flavour profile.

After fermentation, the fermentation broth may optionally be neutralised and/or inactivated and/or homogenised by methods known in the art .

Optionally, if the milk source contains L-rhamnose, it may be heated. This heating step differs from the pasteurisation step and the inactivation step in that it is typically carried out for about 90 minutes at about 90 ⁰ C, in order to develop a caramel -like flavour in the mixture. Without wishing to be bound by theory, it is thought that the sugar sources in the milk basis (lactose, rhamnose etc.) undergo a number of reactions (Maillard reaction pathway) which are responsible for the caramel- like flavour. This heating step may be carried out before or after fermentation.

Optionally, the fermented mixture may be enzymatically treated with a lipase, lactase etc. This may be carried out in order to facilitate the drying step and/or to further modulate the taste and flavour of the final product .

For instance, further enzymatic treatment may provide a vanilla-like, cream-like flavour which imparts a creamy, mouth-coating, long-lasting taste to the final product.

Furthermore, the enzymatic treatment may further improve drying properties of the fermented mixture.

Alternatively also, the fermentation broth may be blended with further ingredients. This may be done prior to drying or prior to cool storage for liquid applications, in order to provide, upon drying or not, a finished product.

As another alternative, a milk source treated in a parallel stream may be added to the fermented broth.

The fermented mixture may then be cooled and the liquid product may be stored at low temperature for further use. Such temperatures are typically between 0 and 10 ⁰ C. The fermented product may then be used as a liquid flavour concentrate in the manufacture of food products.

Alternatively, the fermented mixture is then dried, preferably by spray-drying, to give a milk powder-like product. By milk powder-like products is meant products which have the same appearance as and consistency of milk powder, are based on milk, but are produced in a different way. Thus, the present invention also concerns milk powder-like products.

According to the invention, a milk powder-like product comprising per kg of product l-20000mg, preferably 10- 500mg butanoic acid, 10-lOOOOmg, preferably 100-3000mg hexanoic acid, 10-6000mg, preferably 100-3000mg octanoic acid, 2-13000mg, preferably 50-1000mg decanoic acid is provided.

In an embodiment, the milk powder-like product further comprises per kg of product 0.1-lOmg, preferably 0.1-3mg, more preferably 0.3-lmg of 3 -methyl -butanal, 0.1-lOOmg, preferably 0.1-5mg, more preferably 0.5-2.5mg of 2 -phenyl- acetaldehyde, 0.1-lOOOmg, preferably 0.1-10 mg, more preferably l-5mg of diacetyl, 0.5-1000mg, preferably 0.5- 20mg, more preferably 1-lOmg of acetoin, l-300mg, preferably 1-lOOmg, more preferably 10-70mg of 2,3,5,6- tetramethyl pyrazine, 0.01-50mg, preferably 0.01-20mg, more preferably 0.01-15mg of 2 , 4 , 5-trimethyl oxazole, 0.05-50mg, preferably 0.05-10mg, more preferably 0.1-8mg of 2-phenyl-2-butenal and other phenyl aldehydes (ClO- C16) , 0.1-50mg, preferably 0.1-lOmg, more preferably 0.5- 8mg of benzaldehyde . Preferably, the powder comprises 100- 400mg of butanoic acid, 500-2000mg of hexanoic acid, 500- 1500mg of octanoic acid, 100-700 mg of decanoic acid per kg of product. Such milk powder-like product has a particular honey-cream flavour.

According to another embodiment, the milk powder-like product further comprises per kg of product 0.1-lOOOmg, preferably 0.1-50mg, more preferably l-3mg of diacetyl, 0.5-1000mg, preferably 0.5-50mg, more preferably l-20mg of acetoin, l-300mg, preferably 1-lOOmg, more preferably 15- 80mg of 2 , 3 , 5, 6-tetramethyl pyrazine, 0.01-50mg, preferably 0.01-20mg, more preferably 0.01-15mg of 2,4,5- trimethyl oxazole, 0.1-lOmg, preferably 0.5-8mg of benzaldehyde. Preferably, the powder comprises 50-300mg of butanoic acid, 500-2000mg of hexanoic acid, 400-2500mg of octanoic acid and 100-700mg of decanoic acid per kg of product. Such milk powder-like product has a particular cream-butter flavour.

According to another embodiment, the milk powder-like product further comprises per kg of product 0.1-lOOmg, preferably 0.1-lOmg, more preferably 0.5-5mg of 3 -methyl- butanal, 0.05-100mg, preferably 0.05-10mg, more preferably 0.1-5mg of 2 -methyl -butanal , 0.05-20τng, preferably 0.1- 2mg, more preferably 0.1-lmg of 2 -methyl -propanal , 0.1- 500mg, preferably 0.1-lOmg, more preferably l-8mg of diacetyl, 0.5-500mg, preferably 0.5-20mg, more preferably 1-lOmg acetoin, l-500mg, preferably 1.5-300mg of 2,3,5,6- tetramethyl pyrazine, 0.01-2Omg, preferably 0.01-15mg of 2,4, 5-trimethyl oxazole, 0.05-lOmg, preferably 0.1-8.5mg of 2-phenyl-2-butenal and other phenyl -aldehydes (ClO -16) , 0.1-lOmg, preferably 0.5-8.5mg of benzaldehyde . Preferably, the powder comprises 50-350mg of butanoic acid, 400-2000mg of hexanoic acid, 400-2000mg of octanoic acid and 100-800mg of decanoic acid per kg of product. Such milk powder-like product has a particular cocoa-cream and malt-cream flavour.

According to another embodiment, the milk powder-like product further comprises per kg of product 0.1-20mg, preferably 0.1-lOmg, more preferably 0.5-8mg of 4 -hydroxy- 2 , 5-dimethyl-3 (2H) -furanone . Preferably, the powder comprises 20-400mg of butanoic acid, 300-2000mg of hexanoic acid, 300-2000mg of octanoic acid and 100-800mg of decanoic acid per kg of product. Such milk powder-like product has a particular cream-caramel flavour.

The concentrations of flavour compounds can be determined by head space adsorption to solid phase microextraction fibres, separation by gas chromatography and detection and identification by mass spectrometry. For relative quantification, the response of flavour compounds are

related to the known amounts of the internal standards ethylvalerate (neutral compounds) and 4-methyloctanoic acid (acidic compounds) . It is therefore clear to the skilled person that these values are not absolute values and can slightly vary depending on the type of measurements used.

The milk powder-like products of the invention typically comprise 1-4Og, preferably 1-2Og diglycerides and 0.2-3Og, preferably 0.2-2Og monoglycerides per lOOg of product.

The milk powder-like products obtainable by the present method may be used as such, for ■ example to provide flavoured milk drinks.

The fermented milk source may thus be in the form of a liquid concentrate or a powder.

These milk powder-like products or the liquid fermentation product may be used, according to the present invention, in the manufacture of food products.

Thus, in a final step, the fermented milk source, either as a liquid or a powder, is added to a food product. Typically, the fermented milk source is added to the food product in an amount of 0.05-10% by weight on a dry matter basis. The food product is preferably selected from ice cream, milk powder, chocolate-based product, dairy product, creamer, cocoa beverage, culinary product, infant formula or health care product .

In a preferred embodiment, the taste and flavour modulated milk powder-like products are added in relatively small

amounts to food products, for example 0.05 - 10 wt% for liquid, solid or powder products.

In a particular embodiment of the present invention, the fermented milk source is added to a milk powder or a milk powder precursor. The fermented milk source may be in liquid concentrate or powder form. Thus, the fermented milk source may be added in the powder form to a milk powder and thus provide a flavoured milk powder. Depending on the amount added, the resulting product may have a strong flavour or a more subtle flavour.

In an even more preferred embodiment, the milk powder-like product produced by the present invention is added to a milk powder in an amount of about 0.05-10% by weight, most preferably 1-8% by weight of said milk powder.

Alternatively, the fermented milk source may be added to a milk powder precursor and further spray-dried to yield the flavoured milk powder. By "milk powder precursor" is to be understood a standard liquid milk composition prior to spray-drying .

The resulting products may thus be provided with a subtle, subliminal, distinct flavour perception.

In another embodiment of the present invention, the fermented milk source is added to a frozen dessert mix. The fermented milk source may again be in the form of a liquid concentrate or a powder. It may be added to the frozen dessert mix at any stage of manufacture of the frozen dessert. This results in a frozen dessert, preferably an ice cream, having a subtle natural flavour.

The present methods offer the advantage that it allows a lot of variation as to the components present in the fermented milk source which can further be used for addition in a wide variety of food products . It combines aspects of the food ripening methods known in the art, and adapts it to a new field, notably that of natural flavour powders or natural flavour liquid concentrate. The powders or liquid concentrates thus obtainable may be used in any food application, such as milk drinks, ice cream, chocolate, soups, dairy product, culinary product, infant formulae, health care products etc.

The method further allows to "tailor" the flavours obtained upon fermentation, such that a modulation in the taste of the milk powder-like product, the liquid concentrate or the final food product is possible.

Thus, the present invention provides many options for the variation and modulation of milk powder-like product, whereby variation can be obtained by any combination of amino acid and/or processing conditions as described above. The variety of flavours obtainable by the method of the present invention lends itself thus to a wider range of applications.

Referring to Figures 1, 2 and 5 which represent some embodiments of the methods of the present invention, it can be seen that various flavours, in such cases sweet honey, sweet honey and cream butter, malt and chocolate, and milk, chocolate and honey flavours may be obtained. The parameters which may be altered in each case may be the amino acids, the micro-organism (s) and/or the amount

thereof, for instance more than one source, the order of processing etc.

The milk powders or other food products obtainable by methods of the present invention, present the advantage that they are free of additives and the resulting products are thus "clean label" products. This is in contrast to traditional milk-based powders which require the addition of artificial flavouring in order to modulate their aroma.

Furthermore, the flavours are derived from biological processes, e.g. enzymatic, microbial, and are therefore naturally generated flavour compounds, or "bioflavours" .

It is also within the present invention to modify certain components, or mixtures of components when carrying out the processes of the invention in order to obtain perceptible organoleptic differences in the final powders or liquid concentrates. Any combination of taste and flavour modulated milk powder or liquid concentrates by blending or process combinations are included in this application.

For instance, using a mixture of the amino acids phenylalanine, leucine, isoleucine and valine, a chocolate, malt and honey flavour may be obtained. Also, it is conceivable that by adding a source of rhamnose and phenylalanine to the milk source, and heating the mixture before or after fermentation prior to drying would yield a honey-caramel flavour.

Optionally, flavour compounds can be principally added as complementing additives to the fermented milk source.

However, this is not necessary since the present invention provides a wide range of flavourings which are completely natural .

According to a further embodiment of the present invention a method for flavouring a food product is also provided, which method is similar to that described above, except that it does not require a fermentation step.

Thus, in an alternative embodiment, the present method comprises the first step of adding at least one amino acid to a milk source or adding a protease and/or peptidase to the milk source in order to obtain at least one amino acid from a milk protein. This step and all embodiments are the same as described above. Optionally, L-rhamnose may be added to the milk source.

To the milk source is then added, in a second step, a lipase and/or lactase. These enzymes may be as described above .

The milk source is then heat treated. After heat treatment, the milk source may optionally be dried. These are standard procedures known to a skilled person.

Finally, the dried milk source or the liquid milk source may be added to the food product in order to flavour it. The range of food products, the quantities, are as described above.

Thus, optionally, the fermentation step as described above may be omitted and flavour formation may be focused on enzymatic and heat treatment steps.

Thus, endless variations are possible and are within the grasp of a skilled person.

The present invention is further illustrated hereinafter by means of non-limiting examples.

Examples

Example 1 : sweet honey-cream flavoured milk powder-like product

100kg full cream milk powder is recombined with water to a final concentration of 35% dry matter. 0.1kg of L- phenylalanine is added, the pH value adjusted to 6.7 with potassium hydroxide solution and the mix preheated at 8O ⁰ C and UHT treated at 125°C for 20s. After cooling to 30 ⁰ C, 3kg of Lactococcus lactis starter and 0.07kg commercial lipase enzyme preparation is added to start the fermentation. After 10 hours at 30 ⁰ C, the pH is adjusted to 6.7 with potassium hydroxide solution and 0.9kg commercial lactase enzyme preparation is added and incubated for 3 hours. After cooling to 15°C for intermediate storage, the mixture is heat treated at 80 ⁰ C for 80s, homogenized and spray dried.

Example 2 : cocoa-cream flavoured milk powder-like product

100kg full cream milk powder is recombined with water to a final concentration of 35% dry matter. 0.1kg of L- phenylalanine, 0.1kg alpha-ketoglutaric acid, 0.1kg L- leucine, 0.1kg L-isoleucine, 0.1kg L-valine is added, the pH value adjusted to 6.7 with potassium hydroxide solution and the mix preheated at 80 ⁰ C and UHT treated at 125°C for 20s. After cooling to 3O ⁰ C, 3kg of a mixture of strains of Lactococcus lactis starter and 0.03kg commercial lipase enzyme preparation is added to start the fermentation. After 12 hours at 30 ⁰ C, the pH is adjusted to 6.7 with

potassium hydroxide solution. After cooling to 15 ⁰ C for intermediate storage, the mixture is heat treated at 80 ⁰ C for 80s, homogenized and spray dried.

Example 3 ; analysis of flavour compounds

1.8Og NaCl is weighted into a 20ml crimp top vial and a magnetic stirring bar is added. 5.50 g of sample solution containing 12% dry matter is added. 50 μl of internal standard ethylvalerate solution (45μl ethylvalerate in 100ml water) and 50 μl of internal standard 4- methyloctanoic acid solution (75μl of methyloctanoic acid in 250ml water) is added, followed by closing of the vial. The vial is then placed into a water bath at 65°C using a magnetic hot plate stirrer with contact thermometer at a speed range of about 870 rpm. After an equilibration time of 30min the fibre assembly (SPME Fibre Assemply, 2cm, 50/30μm DVB/CAR/PDMS StableFlex mounted in holder, both manufactured by SUPELCO Beliefonte, USA) is inserted by piercing the septa (depth gauge at 20mm) and by exposure the fibre completely to the headspace above the sample solution. After a sorption time of 30min the fibre is retracted and removed into the fibre assembly and out of the vial . The fibre assembly is immediate injected into the GC injector (depth gauge at 30mm) and the separation is started by exposure of the fibre at the same time. After 5min the fibre is retracted and removed from the injector.

The flavour compounds are separated by gas chromatography on a FFAP capillary column (50m, 0.2mm inner diameter, 0.3 μm coating, Agilent Technologies USA) using helium as carrier gas and a temperature gradient from 40 ⁰ C to 250 ⁰ C.

The separated compounds are detected and identified by- mass spectrometry. Relative quantification is done by calculation of the response of flavour compounds in relation to the known amounts of the internal standards ethylvalerate (neutral compounds) and 4-methyloctanoic acid (acidic compounds) .

Example 4 To 98kg of ice cream mix, 3kg of liquid fermented mix or lkg of fermented milk powder is added. The final ice cream mix is then pasteurized at 80 ⁰ C for 80s, followed by homogenisation, cooling to 3 ⁰ C, ice cream aging, freezing and packaging.