WO2021254943A1

WO2021254943A1 - Umami flavour composition

Info

Publication number: WO2021254943A1
Application number: PCT/EP2021/065922
Authority: WO
Inventors: Elaheh JAMALZADEH; Marco Alexander Van Den Berg
Original assignee: Dsm Ip Assets B.V.
Priority date: 2020-06-17
Filing date: 2021-06-14
Publication date: 2021-12-23

Abstract

The present invention relates to a method for producing an umami flavour composition, said method comprises providing a plant based protein isolate and contacting the plant based protein isolate with a protease, with an exopeptidase and with a glutaminase, to produce the umami flavour composition, wherein the plant based protein isolate does not comprise cereal prolamins.

Description

UMAMI FLAVOUR COMPOSITION

Field

The present invention relates to a method for producing an umami flavour composition. According to another aspect, the present invention relates to an umami flavour composition. Further, the present invention relates to the use of the umami flavour composition.

Background of the invention

Umami is a basic taste which is commonly used in all kind of food items, like soups, sauces, marinades and seasonings. Compositions comprising free glutamate are known for their umami enhancing properties in certain types of food. Flavour composition rich in free glutamate are derived from microorganisms such as yeast. Glutamate is also known as a strong enhancer of umami flavour in combination with 5’-ribonucleotides.

Glutamate, or glutamic acid, is commonly used as food additive and flavour enhancer in the form of its sodium salt, i.e. monosodium glutamate (MSG). The inclusion of MSG in food products requires to label the food products with the E number E621.

Over the years consumers more and more desire clean label food, without E numbers. Preferably the consumers use food wherein only ingredients which are known to the consumer from their own cupboard are included (i.e. consumer recognizable ingredients). Simultaneously, consumers would not accept food items which have less flavour. Hence the food industry is challenged with the provision of clean label food items while providing all the flavour properties as before.

With regard to umami, there is a need in the art to provide umami flavour compositions which are derived from consumer recognizable ingredients. At the same time, the umami flavour composition needs to provide enough umami flavour to food or feed items so the consumer would not be confronted with less flavour. Also, the same umami flavour composition providing enough umami flavour, should not impart a specific off-flavour to the food or feed items. An example of a raw material used for production of umami flavour composition is mushroom. The disadvantage of mushroom based flavour compositions is that they provide mushroom notes to the food or feed product the flavour composition is added to.

Wheat is another promising source for providing an umami flavour. However, the disadvantage of wheat is that it may comprise cereal prolamins. Cereal prolamins, named glutenins and gliadins in wheat, secalins in rye, and hordeins in barley, are the major storage proteins found in the endosperm of cereal grains. The proteins are known allergens able to trigger an IgE response in atopic individuals. Hence, a limiting factor of wheat is that it products derived therefrom might be allergenic. Soy is another promising source for providing an umami flavour. However, the disadvantage of soy is that it may be derived from GM containing materials which requires labelling and is unwanted by consumers.

In view of the above, one of the problems to be solved by the present invention, amongst other problems, is the provision of an umami flavour composition that meets the consumer requirements in clean label, derived from consumer recognizable ingredients, without impairing the flavouring properties, wherein the umami flavour is non allergenic.

Detailed description of the invention

The present invention relates to a method for producing an umami flavour composition, said method comprises providing a plant based protein isolate and contacting the plant based protein isolate with a protease, with an exopeptidase and/or with a glutaminase, to produce the umami flavour composition, wherein the plant based protein isolate does not comprise cereal prolamins.

Surprisingly, the present inventors found that by treatment of prolamin free protein isolate, an umami flavour can be produced that provides similar umami flavour as yeast based umami flavours.

A umami flavour composition is defined herein as a composition which can be safely used in food or feed products for human or animal consumption which composition improves the umami flavour (aroma and taste) perception, umami flavour release and/or mouthfeel of a food composition, by modulation of the flavour (aroma and taste) perception, flavour release and/or mouthfeel of a food composition.

Alternatively, the present invention relates to a method for producing a flavour composition, said method comprises providing a plant based protein isolate and contacting the plant based protein isolate with a protease, with an exopeptidase and with a glutaminase, to produce the flavour composition, wherein the plant based protein isolate does not comprise cereal prolamins.

Alternatively, the present invention relates to a method for producing an umami flavour composition, said method comprises providing a plant based protein source and contacting the plant based protein source with a protease, with an exopeptidase and with a glutaminase, to produce the umami flavour composition, wherein the plant based protein source does not comprise cereal prolamins.

In a preferred embodiment, the present plant based protein isolate does not comprise wheat gluten. More preferably, the present plant based protein isolate does not comprise glutenins and/or gliadins. More preferably, the present plant based protein isolate is not, or is not derived from, wheat and/or cereal.

A protease as used in the present context is defined as a hydrolase acting on peptide bonds. An endoprotease acts on peptide bonds in an endo-fashion, i.e. cleaving the peptide bonds anywhere in the polypeptide chain in contrast to a (exo)peptidase which is defined herein as a hydrolase acting on peptide bonds in a protein substrate in an exo fashion, i.e. acting near the ends of the polypeptide chain. The endoproteases are divided into subclasses on the basis oftheir catalytic mechanism: serine endoproteases (EC 3.4.21 xx), cysteine endoproteases (EC 3.4.22.xx), aspartic endoproteases (EC 3.4.23.xx) and metallo-endoproteases (EC 3.4.24.xx). Preferably, the present endoprotease has EC number 3.4.21 .62.

In a preferred embodiment, the protease in the step of contacting the plant based protein isolate with a protease is an endoprotease. More preferably an endoprotease derived from Bacillus licheniformis. More preferably the endoprotease is a serine endoprotease, preferably a serine endoprotease that comprises subtilisin A or consists of subtilisin A. An example is Alcalase®. The advantage of using the endoprotease is that proteins in the plant based protein isolate are efficiently hydrolysed. Preferably, the present endoprotease has an enzyme activity of 605 Du/g.

Preferably, the exopeptidase is derived from Aspergillus oryzae. More preferably, the exopeptidase is an enzyme mixture comprising endoprotease, preferably derived from Aspergillus oryzae. An example of such an enzyme mixture is Flavourzyme®. Preferably, the enzyme activity of the present enzyme mixture is 1255 U/g. Preferably, the exopeptidase has EC number 3.4.11 .1 .

Glutaminase (EC 3.5.1.2) is an amidohydrolyse enzyme. An example of such an enzyme mixture is GLUTAMINASE SD-C100S derived from Bacillus.

Alternative enzymes, enzyme mixtures or enzyme preparations with similar activity are also in scope of the present invention.

In a preferred embodiment, the present method further comprises contacting the plant based protein isolate with a proline-specific endoprotease. A proline-specific endoprotease is defined herein as an endoprotease cleaving protein or oligopeptides substrates at the C-terminal side of a proline residue in the protein or oligopeptides substrate. The proline-specific endoprotease has been classified as EC 3.4.21 .26. The enzyme can be obtained from various sources such as mammalian sources, bacteria (e.f. Flavobacterium ) and fungi ( Aspergillus , in particular Aspergillus niger). The enzyme of Aspergillus niger has been described in detail in WO02/45524, WO02/46381 , WO03/104382. A suitable fungal enzyme from PeniciUium chrysogenum is disclosed in W02009/144269. A suitable bacterial enzyme from Flavobacterium meningosepticum is disclosed in W003068170. The advantage of using a proline-specific endoprotease is that it reduces any potential bitterness of the present umami flavour composition.

Preferably, the amount of proline-specific endoprotease is within the range of 0.1 to 5% (wt) on dry matter of the flavour composition. More preferably, the amount of proline-specific endoprotease is within the range of 0.5 to 3% (wt) on dry matter of the flavour composition. Preferably, the time of the contacting step is within the range of 0.5 to 24 hours. More preferably within the range of 1 to 5 hours.

In a preferred embodiment, the present plant based protein isolate is derived from a plant belonging to the family of the Fabaceae. In a further preferred embodiment, the present plant based protein isolate is derived from a plant belonging to the genus Vigna, genus Cicer or genus Pisum.

Preferably, the present plant based protein isolate is chosen from the group consisting of mung bean (Vigna radiata), moth bean (Vigna aconitifolia), adzuki bean (Vigna angularis), mungo bean (Vigna mungo), jungle mat bean (Vigna trilobata), rice bean (Vigna umbellate), zompi pea (Vigna vexillata), Sarawak bean (Vigna hosei), hairy cowpea (Vigna luteola), Bambara groundnut (Vigna subterranea), cowpea (Vigna unguiculata), parsnip bean (Vigna lanceolate), beach pea (Vigna marina), Oahu cowpea (Vigna owahuensis).

Preferably, the present plant based protein isolate is chosen from the group consisting of Cicer acanthophyllum, Cicer anatolicum, Cicer arietinum, Cicer atlanticum, Cicer bijugum, Cicer canariense, Cicer chorassanicum, Cicer cuneatum, Cicer echinospermum, Cicer fedtschenkoi, Cicer flexuosum, Cicer graecum, Cicer incisum, Cicer judaicum, Cicer kermanense, Cicer macracanthum, Cicer microphyllum, Cicer mogolatvicum, Cicer montbretia, Cicer multijugum, Cicer oxyodon, Cicer pinnatifidum, Cicer pungens, Cicer rechingeri, Cicer reticulatum, Cicer songaricum, Cicer spiroceras, Cicer stapfianum, Cicer subaphyllum, Cicer tragacanthoides and Cicer yamashitae.

Preferably, the present plant based protein isolate is chosen from the group consisting of Pisum abyssinicum, Pisum fulvum, Pisum sativum, Pisum sativum subsp. elatius and Pisum sativum subsp. sativum.

More preferably, the present plant based protein isolate is derived from a plant selected from the group consisting of mung bean ( Vigna radiata), pea ( Pisum sativum) and chick pea (Cicer arietinum). The present inventors found that using protein isolate derived from mung bean ( Vigna radiata), pea ( Pisum sativum) and chick pea (Cicer arietinum) provides umami flavours similar to yeast based umami flavours. Furthermore, the mung bean (Vigna radiata), pea ( Pisum sativum) and chick pea (Cicer arietinum) do not comprise allergens, or not to an amount that, the derived umami flavour is allergenic. More preferably, the present plant based protein isolate is mung bean (Vigna radiata) or is pea (Pisum sativum). Most preferably, the present plant based protein isolate is mung bean (Vigna radiata). Mung bean is also known as green gram, golden gram, mash bean, green soy or celera- bean.

Alternatively, the present plant based protein isolate is fava bean (Vida faba) or fava bean protein isolate.

Alternatively, the present plant based protein isolate is lentil (Lens culinaris) or is lentil protein isolate.

Alternatively, the present plant based protein isolate is soy bean (Glycine max) or is not soy bean (Glycine max).

Alternatively, the present plant based protein isolate is lupin bean (Lupinus) or lupin protein isolate (Lupinus). In a preferred embodiment, the present method comprises contacting the plant based protein isolate with the protease first and subsequently with the exopeptidase and the glutaminase.

In a more preferred embodiment, the present step of contacting the plant based protein isolate with the exopeptidase and the glutaminase is a single step. Preferably a single step wherein these enzymes are contacting the plant based protein isolate at the same time. Hence, the present contacting the plant based protein isolate with the exopeptidase and the glutaminase is preferably carried out simultaneously. Therefore, the exopeptidase and the glutaminase can be added together or separately, as long as these (active) enzymes are contacting the plant based protein isolate at overlapping times.

In a more preferred embodiment, the present step of contacting the plant based protein isolate with the exopeptidase, glutaminase and proline specific endoprotease is a single step. Preferably a single step wherein these enzymes are contacting the plant based protein isolate at the same time. Hence, the present contacting the plant based protein isolate with the exopeptidase, glutaminase and proline specific endoprotease is preferably carried out simultaneously. Therefore, the exopeptidase, glutaminase and proline specific endoprotease can be added together or separately, as long as these (active) enzymes are contacting the plant based protein isolate at overlapping times.

In a preferred embodiment, the present step of contacting the plant based protein isolate with the protease, exopeptidase and glutaminase is carried out at a pH within the range of 3 to 10 and / or a temperature within the range of 50°C to 70°C.

In a preferred embodiment, the step of contacting the plant based protein isolate with the protease, or with the protease, exopeptidase and glutaminase, is carried out at a pH within the range of 4 to 10 and / or a temperature within the range of 50°C to 70°C. More preferably the pH is within the range of 6 to 9, more preferably within the range of 7.5 to 8.5, such as at pH 8. More preferably the temperature is within the range of 55°C to 65°C. The time of the contacting step can be varied depending on the amount of protease used, which is routine for the skilled person to determine. Preferably, the time of the contacting step is within the range of 0.5 to 24 hours. More preferably within the range of 1 to 5 hours.

Preferably, the amount of protease during the step of contacting the plant based protein isolate with the protease is within the range of 0.1 to 10% (wt) on dry matter of the plant based protein isolate. More preferably the amount of protease during the step of contacting the plant based protein isolate with the protease is within the range of 0.5 to 5% (wt) on dry matter of the plant based protein isolate.

In a preferred embodiment, the present plant based protein isolate has a dry matter content of 1 to 40%, preferably of 5% to 25%. Preferably the plant based protein isolate has a dry matter content within the range of 1 to 30% dry matter, preferably within the range of 2 to 15% dry matter, more preferably within the range of 3 to 10% dry matter.

Preferably, the present step of providing the plant based protein isolate, comprises suspending or mixing the plant based protein isolate in water to provide a suspension of the plant based protein isolate. Preferably the suspension of the plant based protein isolate has a dry matter content within the range of 1 to 30% dry matter, preferably within the range of 2 to 15% dry matter, more preferably within the range of 3 to 10% dry matter.

In a preferred embodiment, the present plant based protein isolate, or the suspension of the plant based protein isolate, is heated towards a temperature within the range of 50°C to 100°C for a time period within the range of 5 minutes towards 300 minutes, before contacting the plant based protein isolate with the protease, the exopeptidase and/or the glutaminase. Heating can be carried out at a temperature within the range of 50°C to 100°C, such as 60°C to 98°C, 70°C to 96°C or 80°C to 95°C. Preferably the heating is carried out for a time period within the range of 5 to 50 minutes, preferably 10 to 30 minutes, more preferably 15 to 25 minutes. The result of heating the plant based protein isolate is that the efficiency of the subsequent enzymatic treatment is increased. Furthermore, enzymes originally present in the plant based protein isolate, if any, are inactivated which reduces undesired effects. Alternatively, heating can comprise heating at 95°C to 115°C for a time period of 1 to 10 minutes, preferably in a continuous process.

In a preferred embodiment, the present method comprises contacting the plant based protein isolate with the protease at a pH within the range of 6 to 10 and/or wherein contacting the plant based protein isolate with the exopeptidase and with the glutaminase is carried out at a pH within the range of 4 to 6.

In a preferred embodiment, the pH of the plant based protein isolate after the step of contacting the plant based protein isolate with a protease is adjusted to a pH lower than 7, preferably lower than 6.5 or lower than 6. For example, the pH is controlled at a pH within the range of 3 to 7, 4 to 6, or more preferably pH 5 to pH 5.5.

Preferably, the present process comprises a heating step to inactivate the added enzymes Preferably, heat treatment can be carried out at a temperature within the range of 50°C to 100°C, such as 60°C to 98°C, 70°C to 96°C or 80°C to 95°C. Preferably the heating is carried out for a time period within the range of 5 to 50 minutes, preferably 10 to 30 minutes, more preferably 15 to 25 minutes. Alternatively, heating can comprise heating at 95°C to 115°C for a time period of 1 to 5 minutes, preferably in a continuous process.

In a preferred embodiment, the umami flavour composition is concentrated to a dry matter content of more than 20%, preferably more than 25%, more preferably more than 35% or even more than 40%, or even more than 60%. Concentration can be carried out with an evaporator, such as with a falling film evaporator or a falling film plate evaporator.

In another preferred embodiment, the present method further comprises formulating the umami flavour composition with a carrier selected from salt, maltodextrin, starch, highly branched amylopectin, cellulose and gum Arabic, and / or spray drying the umami flavour composition to a dry matter content of at least 95%. In a preferred embodiment, the present umami flavour composition is contacted with active carbon or activated charcoal, preferably before or after concentrating the umami flavour composition.

In a further preferred embodiment, the present umami flavour composition comprises an amount of free glutamate of more than 0.1% (wt) on carrier free dry matter of the umami flavour composition. Preferably more than 0.2% (wt), more than 0.3% (wt), more than 0.4% (wt), more than 0.5% (wt), more than 1% (wt), more than 1.5% (wt), of free glutamate on carrier free dry matter of the flavour composition. More preferably, the present umami flavour composition comprises an amount of glutamate within the range of 0.1 to 1.5% (wt) on carrier free dry matter of the umami flavour composition.

In a further preferred embodiment, the present umami flavour composition comprises an amount of free glutamine of less than 2.0% (wt) on carrier free dry matter of the umami flavour composition. Preferably, less than 1.5% (wt), less than 1 .0% (wt), less than 0.5% (wt), less than 0.4% (wt), less than 0.3% (wt), less than 0.2% (wt), less than 0.1% (wt) of glutamine on carrier free dry matter of the flavour composition.

In a further preferred embodiment, the present umami flavour composition comprises a ratio of free glutamate (on carrier free dry matter of the umami flavour composition) to free glutamine (on carrier free dry matter of the umami flavour composition) of more than 2. Preferably, a ratio of more than 3, a ratio of more than 4, a ratio of more than 5, a ratio of more than 6, a ratio of more than 7, a ratio of more than 8, a ratio of more than 9, a ratio of more than 10 of free glutamate (on carrier free dry matter of the umami flavour composition) to free glutamine (on carrier free dry matter of the umami flavour composition). More preferably, the present umami flavour composition comprises a ratio of free glutamate (on carrier free dry matter of the umami flavour composition) to free glutamine (on carrier free dry matter of the umami flavour composition) of more than 15.

In a further preferred embodiment, the present umami flavour composition comprises an amount of pyroglutamate of less than 1 .5% (wt) on carrier free dry matter of the umami flavour composition. Preferably, less than 1.0% (wt), less than 0.5% (wt), less than 0.4% (wt), less than 0.3% (wt), less than 0.2% (wt), less than 0.1% (wt) of pyroglutamate on carrier free dry matter of the flavour composition.

According to another aspect, the present invention relates to an umami flavour composition comprising an amount of free glutamate of more than 1 % (wt) on carrier free dry matter of the umami flavour composition and/or an amount of free amino acids within the range of 10% to 65% (wt) on carrier free dry matter of the umami flavour composition. Preferably, the amount of free amino acids is within the range of 10% to 50% (wt), or 10% to 40% (wt) or 15% to 30% (wt) on carrier free dry matter of the umami flavour composition.

Carrier free dry matter is defined as the dry matter of the umami flavour composition excluding a carrier. Carriers, known as drying aids, are generally used to make a process economically and technically viable and these additives should have a foodstuff status and contribute to increase the glass transition temperature of the mixture. Examples of such carriers are salt, maltodextrin, starch, highly branched amylopectin, cellulose and gum Arabic.

In a preferred embodiment the present umami flavour composition does not comprise cereal prolamins, or does not comprise glutenins and/or gliadins.

The present umami flavour composition has preferably an amount of total amino acids within the range of 50 to 95% (wt) of the flavour composition. More preferably an amount of total amino acids within the range of 53 to 80% (wt) of the flavour composition. Even more preferably an amount of total amino acids within the range of 55 to 70 (wt) of the flavour composition.

In a further preferred embodiment, the present umami flavour composition comprises glycerol. Preferably an amount of glycerol within the range of 1 to 5% (wt) on dry matter. The advantage of including glycerol is enhanced drying properties.

Preferably, the umami flavour composition is in spray dried form. In spray dried form, the flavour composition is stable for at least 12 months during storage at room temperature.

In a preferred embodiment the present umami flavour composition comprises NaCI. More preferably the present umami flavour composition comprises NaCI in the concentration of 1% 40% (wt), more preferably 5%-40% (wt), even more preferably 5%-20% (wt) or even more preferably 1%- 10% (wt).

In a preferred embodiment the present umami flavour composition comprises maltodextrin. More preferably the present umami flavour composition comprises maltodextrin in a concentration of 1 %-50% (wt), more preferably 5%-40% (wt), even more preferably 5%-20% (wt) of the umami flavour composition.

In a preferred embodiment, the present umami flavour composition is suitable to provide or to enhance an umami flavour or umami taste in food or feed. More preferably, the present umami flavour composition is suitable to provide or to enhance an umami flavour or umami taste in food or feed if dosed in an amount of 0.01 to 0.5% (wt) on salt free dry matter of the food or feed. Even more preferably, the present umami flavour composition is suitable to provide or to enhance an umami flavour or umami taste in food or feed if dosed in an amount of 0.05 to 0.3% (wt), or 0.1 to 0.2% (wt), on salt free dry matter of the food or feed. Hence, the present composition is able to provide an umami flavour while it is dosed in small amounts. A further advantage of the present umami flavour composition is that it does not provide off notes to food or feed. Preferably the food or feed item is bouillon, such as vegetable bouillon. Preferably, the food or feed item is selected from the group consisting of meat products, meat hybrid products, meat alternatives, broths, bouillons, soups, sauces, gravies and instant meals.

In a preferred embodiment, the present umami flavour composition comprises an anti-oxidant, for example vitamin C or vitamin E. The advantage of adding an anti-oxidant is to prevent oxidation of the flavour composition. Preferably, the present umami flavour composition is obtainable by a method according to the present invention.

In a preferred embodiment, the present umami flavour composition is packed in a bag of at least 1 kg, preferably at least 2 kg, preferably at least 3 kg, preferably at least 4 kg, preferably at least 5 kg, preferably at least 6 kg, preferably at least 7 kg, preferably at least 8 kg, preferably at least 9 kg, preferably at least 10 kg. More preferably the flavour composition is packed in a bag of 15 to 30 kg, such as a bag of 20 to 25 kg.

In another aspect, the present invention relates to food or feed comprising the present umami flavour composition. Preferably, the food or feed comprises the present umami flavour composition in an amount of 0.01 to 0.5% (wt) on salt free dry matter of the food or feed. Preferably the food or feed comprises the present umami flavour composition in an amount of 0.05 to 0.3% (wt), or 0.1 to 0.2% (wt), on salt free dry matter of the food or feed.

In a further aspect, the present invention relates to the use of the umami flavour composition for providing an umami taste in food or in feed. More preferably, the present invention relates to the use for enhancing an umami taste in food or in feed. Preferably, the present flavour composition provides or enhances an umami taste without introducing off flavours to the food or feed.

Given the umami flavour enhancing properties of the present invention, the invention also relates to the use of the present umami flavour as a replacer of yeast extract or yeast autolysate. The Food Chemical Codex defines a “yeast extract” as follows: "Yeast Extract comprises the water soluble components of the yeast cell, the composition of which is primarily amino-acids, peptides, carbohydrates and salts. Yeast Extract is produced through the hydrolysis of peptide bonds by the naturally occurring enzymes present in edible yeast or by the addition of food-grade enzymes". The Food Chemical Codex defines Autolysed Yeast as follows: "Autoiysed Yeast is the concentrated, not extracted, partially soluble digest obtained from food-grade yeasts. Solubilisation is accomplished by enzyme hydrolysis or autolysis of yeast cells. Autolysed Yeast contains both soluble and insoluble components derived ifom the whole yeast celf'.

The invention is further illustrated in the examples below, wherein reference is made to the figures 1 and 2.

Figure 1 : Results of a 2-AFC test showing the number of times the products are perceived as more umami as yeast extract Gistex LS

Figure 2: Results of a sensory study showing the umami related attributes compared to

Gistex LS

EXAMPLES

Example 1

Production of umami flavour compositions Several protein isolates were tested to produce umami flavour compositions. The protein isolates are shown in table 1. A protein suspension with a dry matter content of 7.5-10.0% was prepared in a small glass reactor. The suspension was heated for 30 min at 95°C. The suspension was incubated at a concentration of 2.4% (v/w) on dry matter for 3 hours at 60°C with Alcalase (Novozymes, Bogsvaerd, Denmark, endoprotease). The pH was controlled at pH 8. At the end of this incubation the pH of the protein suspension was adjusted to 5.3.

To obtain a product with increased umami flavour, the plant protein suspensions were incubated by Flavourzyme (Novozymes, Bogsvaerd, Denmark, endoprotease and exopeptidase) at a dose of 3 wt% on dry matter, a proline-specific endoprotease (as disclosed in WO02/45524) at a dose of 1 wt% on dry matter, and glutaminase (Amano Enzyme Inc., Nagoya, Japan) at a dose of 1 wt% dry matter to convert the free glutamine into free glutamate during 20 hours at 55°C. Next, the enzymes were inactivated by heating at 90°C for 20 minutes. Thereafter, the products were freeze dried. The composition of the provided umami flavour composition products 1 to 8 are shown in table 2.

Table 1 : composition of produced plant protein hydrolysates 1 to 8

Table 2: content umami flavour compositions

Example 2

Production of an umami flavour composition from mung bean: effect of glutaminase Effect of Glutaminase enzyme on release of free glutamate was investigated in mung bean protein derived from Yantai T.Full Biotech Co. Mung bean protein was used to produce an umami flavour composition. Mung bean suspensions was prepared with a dry matter content of 7.5% in two reaction vessels A and B. The suspensions A and B were heated for 30 min at 95°C. Both suspensions A and B were incubated with Alcalase (Novozymes, Bogsvaerd, Denmark, endoprotease) at a concentration of 2.4 wt% on dry matter for 3 h at 60°C. The pH was controlled at pH 8. At the end of this incubation the pH of the mung bean suspension was adjusted to 5.3.

To obtain product A , Mung bean suspension A was incubated with the following enzymes simultaneously: (1) Flavourzyme (Novozymes, Bogsvaerd, Denmark, endoprotease and exopeptidase) at a dose of 3 wt% dry matter and (2) a proline-specific endoprotease (as disclosed in WO02/45524) at a dose of 1 wt% dry matter and (3) Glutaminase (Amano Enzyme Inc., Nagoya,

Japan) at a dose of 1 wt% dry matter to convert the released free glutamine into free glutamate. The Mung bean suspension A was incubated during 20 hours at 55°C.

To obtain product B, Mung bean suspension B was incubated was incubated with the following enzymes simultaneously: (1) Flavourzyme (Novozymes, Bogsvaerd, Denmark, endoprotease and exopeptidase) at a dose of 3 wt% dm and (2) a proline-specific endoprotease (as disclosed in WO02/45524) at a dose of 1 wt%. In this reaction enzyme Glutaminase (Amano Enzyme Inc., Nagoya, Japan) was not added to the suspension B. The Mung bean suspension A was incubated during 20 hours at 55°C.

After incubations obtaining Products A and B, the enzymes were inactivated by heat shock at 95°C for 20 minutes and the products A and B were freeze dried. The composition of the provided umami flavour composition product A and Product B are shown in table 3.

Table 3: Content umami flavour composition from mung bean

Example 3

Evaluation on the umami effect of Umami flavour compositions in water Six flavour compositions out of 8 products mentioned in Table 1 were evaluated by the sensory panel to select the product with the most umami flavour.

The sensory evaluation session was carried out by skilled sensory experts as a 2-alternative forced choice test (2-AFC) where the persons were asked to indicate which product was perceived as more umami in comparison to Gistex LS (yeast extract, DSM). In order to knock out visual cues and the possibility to smell, the samples were given in black cups, the panelists had to wear a nose clip and to wear welding glasses with red glass, and the booths were red lighted. All samples were tasted in 1% solution (10 g/L) in water.

The results are shown in Figure 1. The results indicate that umami flavour compositions derived from wheat gluten, mung bean, and soybean were perceived as having a significant stronger umami level as Gistex LS (Figure 1).

Example 4

Sensory profile of the Umami flavour composition in water

Six flavour compositions mentioned in Table 1 were evaluated by the sensory panel to select the product with the most umami flavour.

The sensory evaluation session was carried out by savory expert panels to obtain a qualitative description of the products for their odour and flavour characteristics. The panelists were asked to write down the most obvious descriptors. This was done under normal tasting conditions (without nose clip, in day light). The experts were asked to write down the most important attributes that could be perceived when tasting the different flavour compositions. All mentioned attributes that were collected were grouped and are represented in Table 4. For wheat gluten, the panellists came up with umami as typical flavour characteristic. Hereafter, pea, soybean, and mung bean were indicated as umami.

Table 4: Attributes that were mentioned (three times or more) as most important attributes.

Example 5

Sensory profile of the umami flavour compositions in vegetable bouillon The flavour compositions were produced using wheat gluten, pea protein, and mung bean product according the method as described in Example 1. The produced flavour compositions were evaluated in vegetable bouillon using flash profiling method.

The panelists had to rank the products for umami related attributes as well as product typical attributes. The following attributes were ranked: umami flavor (fl), sweet-fl, cardboard-fl, bitter-fl, green/grassy-fl, cabbage-fl, mushroom-fl, salt-fl, and fullness mouthfeel (mf). The flavour compositions were compared with Gistex LS (yeast extract, DSM, the Netherlands), therefore this product was included in the test. The products were given simultaneously to the panellists, who had to rank these products from least to most intense for that attribute. After three attributes the panelists got a new set of products to avoid cooling down of the products. The results were acquired by means of EyeQuestion software, and the results were analysed by using the Friedman testto investigate whetherthe products were different and simultaneously a post hoc test (Nemenyi) was done to show the difference between the products when an attribute showed a statistically significant difference.

To show the effect of the applied processing, the characteristic sensorial profile of the flavour compositions was compared to the yeast extract Gistex LS (DSM, the Netherlands) in vegetable bouillon. The application formulations which were used, are listed in Table 5.

Table 5: Composition of vegetable bullion containing flavour composition in flash profiling

The powder formulations were mixed with tap water (1.5% in solution equivalent to 15 g/L) of 95°C and stirred until homogeneity. The vegetable bouillons were cooled down until 60°C before sensorial evaluation.

In Figure 2 the bar chart with the sum of ranks is given. Among many attributes, umami flavour, sweet flavour, and bitter flavour showed a statistically significant difference. The umami flavor perception was highest for the wheat gluten. Moreover, the mung bean umami flavour also showed an umami perception that was significantly higher than Gistex LS. The umami perception of the pea umami flavour was equal to Gistex LS. The least sweet perceived product was the pea umami flavour, all other products had a comparable sweetness. For the attribute bitterness no significant differences were perceived. Similar letters show no significant difference. Example 6

Flavour impact the umami flavour compositions in tomato soup

The flavour compositions were produced using wheat gluten and mung bean product according the method as described in Example 1. The produced flavour compositions were evaluated in a tomato soup by group of six savoury experts. They received a reference without a flavour composition and two samples with 0.37% of one of the flavour compositions. They received all three samples, were asked individually to describe the product characteristics of each product and afterwards there was a discussion to come to consensus of the outcome. The application formulations which were used, are listed in Table 6.

Table 6: Composition of tomato soup containing flavour composition

The soups containing umami flavour compositions were described as having an increased impact on fullness, richness and depth of the soup. Next to that they increased the body and complexity.

The Wheat Gluten flavour composition was found to be slightly stronger on fullness and created a slightly more balanced tomato soup compared to the mung bean. Example 7

Flavour impact the umami flavour compositions in beef gravy

The flavour compositions were produced using wheat gluten and mung bean product according the method as described in Example 1. The produced flavour compositions were evaluated in a beef gravy by group of six savoury experts. They received a reference without a flavour composition and two samples with 0.37% of one of the flavour compositions. They received all three samples, were asked individually to describe the product characteristics of each product and afterwards there was a discussion to come to consensus of the outcome. The application formulations which were used, are listed in Table 7.

Table 7: Composition of beef gravy containing flavour composition

The gravy’s containing umami flavour compositions were described as having an increased richness, more rounded and complex flavour profile. Next to that the meaty notes in the gravy were enriched and the body and mouthfeel was increased. Both flavour compositions were comparable on effect of the application. Example 8

Flavour impact the umami flavour compositions in white asparagus soup

The flavour compositions were produced using wheat gluten and mung bean product according the method as described in Example 1. The produced flavour compositions were evaluated in a white asparagus soup by group of six savoury experts. They received a reference without a flavour composition and two samples with 0.36% of one of the flavour compositions. They received all three samples, were asked individually to describe the product characteristics of each product and afterwards there was a discussion to come to consensus of the outcome. The application formulations which were used, are listed in Table 8.

Table 8: Composition of white asparagus soup containing flavour composition

The soups containing umami flavour compositions were described as having an increased richness, making the profile of the soup more full and rounded. Next to that it raises the asparagus notes in the soup and they delivered more body and mouthfeel were enriched and the body and mouthfeel was increased.

The Wheat Gluten flavour composition was found to increase the asparagus note more and provided more fullness than the mung bean flavour composition. Example 9

Flavour impact the umami flavour compositions in mushroom sauce

The flavour compositions were produced using wheat gluten and mung bean product according the method as described in Example 1. The produced flavour compositions were evaluated in a mushroom sauce by group of six savoury experts. They received a reference without a flavour composition and two samples with 0.35% of one of the flavour compositions. They received all three samples, were asked individually to describe the product characteristics of each product and afterwards there was a discussion to come to consensus of the outcome. The application formulations which were used, are listed in Table 9.

Table 9. Composition of mushroom sauce containing flavour composition

The sauces containing umami flavour compositions were described as enriching the overall taste and bringing a fuller taste profile. Next to that they improved the balance and complexity of the sauces and increased the mouthfeel and body.

Both flavour compositions were comparable on effect of the application.

Claims

1 . Method for producing an umami flavour composition, said method comprises providing a plant based protein isolate and contacting the plant based protein isolate with a protease, with an exopeptidase and with a glutaminase, to produce the umami flavour composition, wherein the plant based protein isolate does not comprise cereal prolamins.

2. Method according to claim 1 , wherein the plant based protein isolate does not comprise wheat gluten.

3. Method according to claim 1 or claim 2, further comprising contacting the plant based protein isolate with a proline-specific endoprotease.

4. Method according to any of the preceding claims, wherein the plant based protein isolate is derived from a plant belonging to the family of the Fabaceae.

5. Method according to any of the preceding claims, wherein the plant based protein isolate is derived from a plant belonging to the genus Vigna, genus Cicer or genus Pisum.

6. Method according to any of the preceding claims, wherein the plant based protein isolate is derived from a plant selected from the group consisting of mung bean ( Vigna radiata), pea ( Pisum sativum) and chick pea ( Cicer arietinum), preferably mung bean ( Vigna radiata).

7. Method according to any of the preceding claims, comprising contacting the plant based protein isolate with the protease first and subsequently with the exopeptidase and the glutaminase.

8. Method according to any of the preceding claims, wherein the step of contacting the plant based protein isolate with the protease, exopeptidase and glutaminase is carried out at a pH within the range of 3 to 10 and / or a temperature within the range of 50°C to 70°C.

9. Method according to any of the preceding claims, wherein the plant based protein isolate has a dry matter content of 1 to 40%, preferably of 5% to 25%.

10. Method according to any of the preceding claims, wherein the plant based protein isolate is heated towards a temperature within the range of 50°C to 100°C for a time period within the range of 5 minutes towards 300 minutes, before contacting the plant based protein isolate with the protease, the exopeptidase and the glutaminase.

11. Method according to any of the preceding claims, wherein contacting the plant based protein isolate with the protease is carried out at a pH within the range of 6 to 10 and/or wherein contacting the plant based protein isolate with the exopeptidase and with the glutaminase is carried out at a pH within the range of 4 to 6.

12 Umami flavour composition comprising an amount of free glutamate of more than 1% (wt) on carrier free dry matter of the umami flavour composition, and an amount of free amino acids within the range of 10% to 65% (wt) on carrier free dry matter of the umami flavour composition.

13. Umami flavour composition according to claim 12, wherein the umami flavour composition is obtainable by the method according to any of the claims 1 to 10.

14. Food or feed comprising the umami flavour composition according to any of the claims 11 to 13.

15 Use of the umami flavour composition according to any of the claims 11 to 13, for providing an umami flavour in food or feed.