FIELD OF THE INVENTION The present invention relates to an oat base with enhanced viscosity and to products made therefrom, such as non-dairy yogurt. The invention also relates to corresponding processes of manufacture . BACKGROUND OF THE INVENTION

Oat drinks ("oat milk") for use as cow milk substitutes (EP 731646 Al ; US 5686123; US 6451369 A) and as a raw material for other non-dairy milk products are known in the art. They are preferred by many customers for various reasons, such as for their content of soluble β-glucan fiber beneficial to health, their lack of potentially allergenic proteins and of lactose, which cannot be digested by the majority of the global

population.

In prior art processes for preparing oat milk the starting material such as oat flour or oat bran is heated to a temperature and for a time sufficient to substantially prevent the

development of endogenous enzymatic activity, in particular lipase/lipoxygenase activity, but also β-glucanase activity. Oat milk known in the art may be also be termed "oat base" since, in addition to its use as a drink, in particular milk drink, it can be used as a base for the manufacture of other food products, such as oat yogurt, oat cream or oat batter, or be used as a food additive .

Due to the low fat content of oat milk (typically 0.5 % by weight) fat in form of vegetable oil, such as rapeseed oil, is often added to the product .

Oat base or milk can be used for preparing oat drinks by addition of sugar, flavorings, fruit extracts, etc. It is desirable for those drinks to be similar in taste, texture, and viscosity to corresponding drinks prepared from cow milk. This is not problematic as far as drinks of low viscosity, that is, of a viscosity similar to dairy milk, are concerned. A problem however arises in the use of oat base for the preparation of products corresponding to cow milk products of enhanced viscosity such as yogurt or sour cream. Cow milk products of enhanced viscosity like yogurt or sour cream are prepared by fermentation using cultures of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus bacteria as such or in combination with other lactobacilli and bifidobacteria. Decrease of pH by lactic acid formed in the process coagulates milk protein, thereby substantially increasing the viscosity of the product.

Oat base and cow milk have a protein content of about 1 % by weight and about 3.5 by weight, respectively.

WO 2014/123466 Al discloses a deamidated liquid oat base and a process for its manufacture.

Milk protein crosslinking and/or modifying enzymes such as transglutaminase, laccase, tyrosinase, peroxidase, sulfhydryl oxidase and protein glutaminase are used in the dairy industry to stabilize the structure of milk-based products. WO 2007/060288 Al discloses a method of manufacturing soured fresh dairy products by the combined use of transglutaminase, an enzyme activating compound with oxidative or reducing properties such as yeast extract and glutathione, and an acidifying agent in form of a microbiological starter culture, a chemical acidifying agent and mixtures thereof, enabling compensating for loss of viscosity in the production of yogurt when using milk of a lower protein content. WO 2013/064736 Al discloses a liquid enzyme formation comprising a polyol -water suspension of a milk protein

crosslinking and/or modifying enzyme such as transglutaminase. OBJECTS OF THE INVENTION

It is an object of the invention to provide an oat drink or base of the aforementioned kind of substantially enhanced viscosity.

It is another object of the invention to provide products of enhanced viscosity obtainable from said oat base or drink by fermentation. It is a further object of the invention to provide

corresponding processes.

Further objects of the invention will be evident from the following summary of the invention, the description of preferred embodiments thereof, and the appended claims.

SUMMARY OF THE INVENTION

According to the present invention is provided a modified aqueous oat base or drink, in the following referred to as

"modified oat base" of the aforementioned kind of substantially enhanced viscosity. The viscosity of the modified oat base is obtained by enzymatic means.

According to an important aspect of the invention the viscosity of the modified oat base is not obtained by adding one or more viscosity modifying agents or thickeners, that is, agents forming highly viscous aqueous solutions such as xanthan gum, sodium alginate, hydroxypropyl methyl cellulose, carrageenan, agar, gelatin, and the like.

The term oat base as used herein relates to aqueous liquid oat products of low viscosity and having a high content of maltose obtained or obtainable from oats by treatment with starch-degrading enzymes, such as a- and/or β-amylase. The term oat drink as used herein relates to ready-to-drink, pasteurized oat base, which may contain additives which do not substantially affect the viscosity thereof, such as sodium chloride, vegetable oil such as, for instance, rapeseed oil, sweetener, vitamin, and flavoring .

The viscosity enhanced oat base or drink of the invention is obtained by cross- linking oat protein comprised by the oat base or drink supplemented by vegetable protein isolate, in particular by denaturated vegetable protein isolate.

According to the invention protein comprised by the protein supplemented oat base or drink is crosslinked by means of transglutaminase .

According to another important aspect of the invention the oat base is a deamidated oat base. According to a preferred aspect of the invention deamidation is carried out in the presence of calcium carbonate.

According to a further preferred aspect of the invention the added vegetable protein isolate consists of or comprises

denaturated vegetable protein isolate, in particular thermally denaturated vegetable protein.

Preferred vegetable protein isolates according to the invention comprise pea protein isolate, potato protein isolate, faba bean protein isolate, chickpea protein isolate, lentil protein isolate, pea protein isolate being particularly

preferred.

According to the present invention is disclosed a process for producing a non-dairy product of enhanced viscosity,

comprising providing a mixture of deamidated oat base or drink and vegetable protein isolate, in particular denaturated

vegetable protein isolate; crosslinking glutamine and lysine units of the vegetable protein isolate by means of

transglutaminase. It is preferred for the mixture of deamidated oat base or drink to be heated to a temperature of 80° or more prior to the addition of transglutaminase.

In particular, according to the present invention is disclosed a process for producing an oat base of enhanced viscosity, comprising: providing an aqueous oat base or drink optionally comprising a viscosity promoting agent; providing a native vegetable protein isolate and/or a denaturated vegetable protein isolate; combining the aqueous oat base or drink and the vegetable protein isolate; heating the combination of aqueous oat base or drink and protein isolate to a temperature of 80 °C or more and keeping it at this temperature for a time sufficient to form a combination of aqueous oat base or drink and heat-treated vegetable protein isolate; bringing the combination of aqueous oat base or drink and heat-treated vegetable protein isolate to a temperature of from 35 °C to 65 °C; adding a protein cross- linking amount of transglutaminase to the combination of aqueous oat base or drink and heat-treated vegetable protein isolate while keeping the mixture at said temperature of from 35 °C to 65 °C for a time required to increase viscosity by a factor of 2 or more, in particular of 5 or more, to form a viscosity enhanced oat base or drink; inactivating transglutaminase by heating the viscosity enhanced oat base or drink to a temperature of 80 °C or more, in particular of 90 °C or more; cooling the viscosity enhanced oat base or drink to one of : room temperature or below if desired to be stored for a longer time or to a temperature suitable for further processing; with the proviso that protease activity is excluded from the process.

Also disclosed is an oat base or drink of enhanced viscosity obtainable the process of the invention and its use as a food, a food additive or a starting material for production of a food, all intended for human consumption, in particular for the production of yogurt, sour cream, and fresh cheese.

The invention is based on the insight that the vegetable protein isolate is to be used preferably in denaturated form and to be additionally denaturated by heat treatment during

processing. A most preferred vegetable protein isolate is a pea protein isolate, in particular in denaturated form.

A preferred amount of added vegetable protein is from 0.5 % or 1 % by weight to 2 % by weight or more, such as to 3 % by weight or more, and even up to 4 % or 5 % or 7 % by weight or more .

A preferred aqueous oat base of the invention has a dry matter content of from 7 to 15 % by weight, preferably of about 10 % by weight, the oat base dry matter comprising from 10 % to

50 % by weight of maltose or of a mixture of maltose and glucose, from 30 % to 80 % by weight of maltodextrin.

Preferred heat treatment conditions comprise the combination of a denaturation temperature of from 80 °C or more, in

particular of 90 °C or more, in particular of about 95 °C, in combination with a denaturation period of from 5 min to 60 min, in particular of from 10 min to 30 min.

Dairy yogurt has viscosity of from about 2000 Pa-s (stirred yogurt) to about 5000 Pa-s (Greek and Turkish yogurt) . A process of the invention enhances viscosity by factor of 2 or 5 or more and even by a factor of 10 or more, such as to obtain a viscosity of 10000 Pa-s or more and even of 20000 or more, which corresponds the viscosity of curd. In comparison the viscosity of a corresponding fermented product obtained by omitting protein cross- linking in a process of the invention is below 1000, typically in the range of 50 Pa-s - 500 Pa-s.

Preferred vegetable protein isolates contain about 14 % by weight or more, more preferred 15 % or more, such as about 17 % and even up to 20 % by weight of glutamine/glutamic acid and about 5 % or more, more preferred about 6 % by weight or more, such as about 7 % or 8 % by weight of lysine in combination. Pea (Pisum sativum L.) protein such as in form of pea protein isolate is the most preferred protein for use in the process. Pea protein enhances viscosity substantially more than other vegetable proteins investigated, such as lentil, faba bean, potato, hemp, canola, and oat protein. Preferred vegetable proteins have an isoelectric point of from about 4.0 to about 5.0, in particular of from about 4.2 to 4.6, most preferred of about 4.4.

A preferred amount of added transglutaminase is from 0.1 U/ to 5 U/g, in particular from 0.2 U/g to 2 U/g, most preferred about 1 U/g.

In addition to the enhancement of viscosity a process of the invention enhances elastic modulus such as from below 100 Pa to 500 Pa or 1000 Pa or more.

Furthermore a product of a process of the invention has higher yield point, in particular one higher by a factor of 1.5 or 2 or more, the yield point being the strain point at which the network of the gel breaks. The process of the invention thus provides products with improved gel properties, that is, stronger gels .

A viscosity promoting agent of the invention is one that does not raise or does only raise insignificantly, such as by a factor of less than 1.5 or less than 1.2 or 1.1, the viscosity of an aqueous media in which it is dissolved at a concentration of up to 5 % by weight. A preferred viscosity promoting agent is calcium carbonate. Another preferred viscosity promoting agent is a combination of magnesium carbonate and calcium carbonate.

It is preferred for the cereal oats suspension to be heated for protein crosslinking to a temperature of from about 50 °C to about 75 °C for a time sufficient to produce an oat base of desired composition while controlling its viscosity by varying the amount and/or kind of added vegetable protein and/or the amount of transglutaminase .

According to the invention such control is also possible by using a combination of vegetable protein from different sources, such as, for instance, combinations of pea protein isolate, potato protein isolate, faba bean protein isolate, chickpea protein isolate, lentil protein isolate.

The oat base or drink of the invention of improved viscosity and products made from it differ from prior art oat bases and corresponding products made therefrom by their substantially enhanced viscosity. The use of transglutaminase in the process of the invention has no or only insignificant effect on the taste of the products.

According to a preferred aspect the viscosity enhanced oat base or drink of the invention can be used for the production of non-dairy cheese.

According to the invention the oat base or drink of enhanced viscosity produced by process of the invention can be employed as starting material in a fermentation process for the manufacture of a fermented non-dairy product such as yogurt and cheese, the process comprising: bringing the oat base or drink of enhanced viscosity to a fermentation temperature of from about 35 °C to about 50 °C, in particular of about 43 °C; adding a fermentation starter culture, in particular a yogurt or cheese starter culture; maintaining the cereal suspension or oat base of enhanced viscosity at the fermentation temperature for a time period sufficient to form a fermented product, in particular a non-dairy yogurt-like or cheese-like product, such as for a time period of from 8 h to 24 h, in particular of from 12 h to 15 or 18 h.

According to a preferred aspect of the invention a minor amount of sugar, such as from 0.2 % by weight or 0.5 % by weight or 0. 7 % by weight and up to 1 % by weight of dry matter, is added at the start or during the process for nourishment of the yogurt culture. According to another preferred aspect of the invention a buffering agent is added at the start or during the process to stabilize H, in particular at a pH of less than 6.0, in particular of less than 5.0, most preferred at about 4.5.

It is preferred not to agitate the cereal suspension by stirring during fermentation so as to not jeopardize the gellous nature of the product .

Furthermore, according to the present invention is disclosed a process for producing a fermented non-dairy product of enhanced viscosity, comprising: providing deamidated oat base or drink optionally comprising a viscosity promoting agent; providing native and/or denaturated vegetable protein isolate; mixing the oat base or drink and the protein isolate; heating the mixture of oat base or drink and protein isolate to a temperature of 80 °C or more, in particular of 90 °C or more, most preferred of about 95 °C for from 5 min to 60 min, in particular from 10 min to 30 min; cooling the mixture; adding transglutaminase and a

fermentation culture to the mixture; keeping the mixture

comprising transglutaminase and fermentation culture at a fermentation temperature, such as at a temperature of from 35 °C to 50 °C, in particular of about 43 °C, for a time period sufficient for forming a fermented product, in particular a yogurt-like or curd- like product, such as for a time period of from 8 h to 24 h, in particular of from 12 h to 15 or 18 h;

optionally destroying enzymatic activity by heating the

fermentation product; cooling the fermented product to room temperature or below.

According to a preferred aspect of the invention, the pH of a process of the invention is controlled by, prior to incubation with transglutaminase, adjusting and/or holding the mixture of deamidated oat base and vegetable protein isolate at a pH substantially higher than that of the isoelectric point of the vegetable protein isolate, such as higher by 1.0 pH units, in particular higher by 1.5 pH units or 2.0 pH units or more.

Preferred fermented products obtainable by the process are non-dairy yogurt, non-dairy sour cream, and non-dairy fresh cheese . In the following the invention will be described in more detail by reference to a number of preferred embodiments thereof.

DESCRIPTION OF PREFERRED EMBODIMENTS

Material and Methods

Oat kernels: Dehulled, steam treated, wet ground or dry ground .

Oat bran (Frebaco Kvarn AB, Lidkoping, Sweden) : Prepared from steam treated Swedish oat grain by grinding in a rolling mill. Composition (% by weight) : Protein 18, fat 7, carbohydrate 45, fiber 16 %, water 9.5.

Enzymes: Transglutaminase, alpha-amylase, beta-amylase as well as yogurt and fresh cheese products, such as sour cream, quark and cream cheese, are available from various commercial sources such as Danisco DuPont (Yomix™, Denmark) and Christian Hansen (YoFlex™, eXact™; Denmark) . Prior to use transglutaminase and yogurt culture were dissolved in tap water (15 U/g and 0.001- 0.2 g/g, respectively).

Alpha -amylase activity: One Ceralpha unit is defined as the amount of enzyme required to release one micromole of

p-nitrophenol from BPNPG7 (non-reducing end blocked p-nitrophenyl maltoheptaoside) in one minute under defined assay conditions: http : //secure . megazyme . com/files/BOOKLET/K-BETA3_1010_DATA. pdf Beta-amylase activity: One ΒΝΡβ-Θ3 (p-nitrophenyl -β-D- maltotrioside) unit is defined as the amount of enzyme required to release one micromole of p-nitrophenol from ΡΝΡβ-Θ3 in one minute under defined assay conditions:

http://secure.megazyme.com/files/BOOKLET/K-BETA3 1010 DATA. pdf.

Transglutaminase activity: Determined by the hydroxamate method. One unit corresponds to the amount of enzyme generating 1 μπιοΐ of hydroxamic acid per min at pH 6.0.

Viscosity: Measured with Kinexus Pro+, Malvern Instruments, UK; http://www.malvern.com/en/producs/product-range/kinexus- range/kinexus-por-plus/ . EXAMPLE 1. Process for producing the viscosity-improved oat base of the invention from a state -of-the art oat base

Materials: Oat base produced according to EP 1124441 Al

containing about 1 % by weight of oat protein, to which rapeseed oil had been added to rise the total fat content to about 3 % by weight; transglutaminase ACTIVA TG-WM™ (100 U/g) , Aj inomoto (Japan) ; pea protein, Nutralys™ S85F and F85F containing about 8 % by weight of protein (Roquette, France) ; YoMix™ 511 yogurt culture, Danisco DuPont (Denmark) ; rapeseed oil (AAK, Sweden) ; granulated sugar. Aqueous solutions were prepared for addition during fermentation: a) 15 U/g transglutaminase solution b) 1.8 by weight Yomix™ 511 solution. Pea protein (3.8 g) was added to 94 g oat base in a glass flask and the mixture shaken to disperse the protein. The flask was immersed in a boiling water bath of 95 °C and kept at this temperature for 10 min, then cooled to room temperature by immersing it in cold water. The mixture was used as starting material .

First variety of the process of the invention: Simultaneous cross-linking and fermentation. The following components were admixed with the starting material: 265 μΐ transglutaminase solution (1 U/g protein); 340 μΐ YoMix solution (0.0006 %) ; 1.15 g rapeseed oil (to make total fat content about 4 % by weight) ; 0.69 g of granulated sugar. The mixture was shaken to mix and dissolve the components, then kept in a water bath of 43 °C overnight. The reaction was quenched by bringing the reaction mixture to boiling in a microwave oven, followed by shaking and renewed heating so as to bring it to the boiling point. The mixture was cooled and stored in a refrigerator for analysis.

Second variety of the process of the invention: Consecutive cross-linking and fermentation. Transglutaminase solution (265 μΐ, 1 U/g protein) was added to the starting material (about 100 g) . The mixture was stored for 3 h in a water bath of 50 °C and agitated once hourly. The reaction was quenched by bringing the reaction mixture to boiling in a microwave oven followed by shaking and renewed heating so as to bring it to the boiling point in the oven. The cooled product is a viscosity enhanced oat base or oat drink of the invention. It can be used as such as a food product or in the manufacture of food products or be fermented. If used for fermentation, the following agents were added to the cooled viscosity enhanced oat base or oat drink: 340 μΐ YoMix solution (0.006 %) , 1.15 g rapeseed oil to make total fat content about 4 % by weight); 0.69 g of granulated sugar (0.69 % by weight) ; it is however within the ambit of the invention to add rapeseed oil and/or granulated sugar at an earlier stage of the process. The mixture was incubated overnight in a water bath of 43 °C. The so produced non-dairy yogurt was cooled and kept in a refrigerator for analysis.

EXAMPLE 2. Crosslinking efficacy of different protein isolates

A number of vegetable protein isolates were included to demonstrate their crosslinking properties at a total protein content of 4 % by weight in deamidated oat base. The results were confirmed by rheology measurements, illustrated in Table 1

(consecutive crosslinking and fermentation) and Table 2

(simultaneous crosslinking and fermentation) . Oat base without added protein isolate resulted in zero shear viscosity of about 50 Pa-s, elastic modulus (G' ) of about 5 Pa and a yield point of about 1.

Table 1. Crosslinking effect of transglutaminase on fermented products of combinations of protein isolates and deamidated oat base, consecutive crosslinking and fermentation

Protein isolate added to a Viscosity, Elastic modulus Yield point

content of 4 % total protein Pa s Pa' Pea (Nutralys S85F) 2 000 300 0.9

Chickpea (FCPP40) 3 000 200 1.1

Pea (Pisane C9) 8 000 800 1

in combination with

deamidated oat drink Table 2. Crosslinking effect of transglutaminase on fermented products of combinations of protein isolates and deamidated oat base, simultaneous crosslinking and fermentation

Protein isolate added to a Viscosity, Elastic modulus Yield point

content of 4 % total protein Pa- s Pa

Pea (Nutralys S85F) 6 000 800 1.9

Potato Solanic 200) 3 000 300 0.9

Faba bean (Vitessence 3600) 18 000 1 200 2.2

Lentil (Vitessence 2550) 2 000 300 1.4

Chickpea (FCPP40) 4 000 300 1.6

Pea (Pisane C9) 40 000 5 000 2

in combination with

deamidated oat drink

EXAMPLE 3. Crosslinking efficacy of different starting materials An important factor for crosslinking properties is the nature of the protein isolate, as demonstrated in Example 2.

Another important factor is whether a protein isolate had undergone a heat treatment prior to crosslinking, as illustrated in Table 3. The effect of heat treatment seems however not to be of a general nature but limited to certain protein isolates, for example pea protein isolate. According to the invention

properties of the resulting gel or curd can be adjusted by varying time and/or temperature of heat treatment . Higher temperatures and/or longer periods of treatment result in increased viscosity and gel strength.

The inclusion of deamidated oat base or oat drink and addition of a viscosity promoting agent are additional factors affecting crosslinking properties and viscosity. The combination of oat base and pea protein that had not been heat-treated prior to transglutaminase incubation did not thicken, while heat treatment resulted in slightly increased viscosity. Table 3. Crosslinking effect of transglutaminase on fermented products of heat-treated (10 min, 95°C) and non-heat-treated combinations of protein isolates and deamidated oat drink, consecutive/simultaneous crosslinking and fermentation

Protein isolate Viscosity, Pa s Elastic modulus, Pa Yield point

Consecutive

Pea (Nutralys S85F) Heated 2 000 200 1

Pea (Nutralys S85F) Non-heated 300 10 1

Potato (Solanic 200) Heated 5 000 1 800 1.5

Potato (Solanic 200) Non-heated 7 000 600 0.6

Simultaneous

Pea (Nutralys S85F) Heated 17 000 800 1.6

Pea (Nutralys S85F) Non-heated 4 500 550 1.6

Potato (Solanic 200) Heated 1 000 1 700 1.1

Potato (Solanic 200) Non-heated 2 000 1 700 0.9

Application of the same treatment to the combination of deamidated oat drink and pea protein in absence of heat -treatment resulted in a somewhat thick gel while heat -treatment provided a strong gel or curd. The effects of deamidated oat drink were confirmed by rheology measurements, illustrated in Table 4. The inclusion of calcium carbonate (corresponding to a calcium

content of about 120 mg/100 ml) as viscosity promoting agent to the oat base increased viscosity and provided an even stronger gel or curd.

Table 4. Crosslinking effect of transglutaminase on fermented products of combinations of pea protein isolate and oat drinks, consecutive/simultaneous crosslinking and fermentation

Oat drink Viscosity, Pa s Elastic modulus, Pa Yield

Consecutive

Deamidated Heated 2 000 200 1

Non-heated 300 10 1

Non-deamidated Heated 600 90 1

Non-heated 40 1 0.9

Simultaneous

Deamidated Heated 17 000 800 1.6

Non-heated 4 500 550 1.6

Non-deamidated Heated 5 500 500 1.4

Non-heated 150 10 1 EXAMPLE 4

Adjusting/holding the pH to/at a pH substantially different from that of the isoelectric point during incubation with transglutaminase increased viscosity and provided a stronger gel or curd.

The isoelectric point of vegetable protein isolates is at about pH 4.5; for pea protein isolates is at about pH 4.4.

Table 5 illustrates the increase in viscosity upon adjusting the pH of oat drink comprising pea protein isolate to pH 6.5 and 7.5 prior to incubation with transglutaminase and fermentation.

Aqueous sodium hydroxide and hydrochloric acid were used for pH adjustment . Table 5. Crosslinking effect of transglutaminase on fermented products with pH adjusted different from isoelectric point, consecutive crosslinking and fermentation.

Oat drink pH Viscosity, Pa.s Elastic modulus, Pa Yield point

Deamidated 6.5 2 000 50 0.9 Deamidated 7.5 3 000 200 1

Non-deamidated 6.5 200 1 0.5

Non-deamidated 7.5 1 500 200 1