Field of the invention

The present invention relates to fermented protein products based on proteinaceous pulse sources having added plant material comprising phytochemicals as well as a method for their preparation and use in a food or a feed.

In particular it relates to fermented protein products based on pulse sources containing plant material comprising phytochemicals in an amount sufficient to increase the functional properties and improving the overall production parameters when used in food or feed.

Background of the invention

It is well known that the continued increase in world population results in a growing need for protein products suitable for human food and animal feed. Food products based on plants do generally have a high nutritional value and they are also relatively cheap.

It is also well known that plant proteins of high quality can be derived from certain crops, such as proteinaceous pulse sources. However, such crops frequently contain harmful and/or bad tasting components and they are therefore unsuitable for food and feed purposes unless they are subject to some degree of processing or treatment. In the following soybeans will be described but the problems associated with soybeans also apply for other types of pulses. Raw soybeans contain trypsin inhibitors. Trypsin inhibitors reduce the activity of trypsin, an essential enzyme in the digestive process. The nutritional value of the raw soy based product is thus relatively low. A heating process can reduce the content of the trypsin inhibitors. However, the heating process might also lead to a reduced nutritional value of the soy protein. Raw soybeans furthermore contain proteins with antigenic properties, e.g. glycinin and β-conglycinin. These proteins are known to induce serious allergic reactions, especially in young mammals where the digestive tract is not yet fully developed. It is known that such proteins can be inactivated by extraction with water-alcohol mixtures and/or by heating. Denaturation of these proteins will, however, result in products with reduced ability to bind water and fat. Overheating may furthermore impair the nutritional value.

The amount and nature of carbohydrates furthermore influence the nutritional value, a-1 ,6-galactosides do e.g. induce flatulence and diarrhea. These compounds can be reduced or eliminated by extraction with water, degradation by the action of specific enzymes or fermentation.

Furthermore, the flavor and taste of the product is essential for the nutritional benefit of such a product. Crude protein products based on pulses are associated with an unpleasant "beany" taste due to the presence of compounds such as 1 -pentanol, 1 -hexanol and 1 -octen-3-ol. These compounds can be reduced by e.g. heat treatment and/or extraction with water/alcohol mixtures.

Any product processing is also associated with increased production costs.

Protein products with improved flavor are commercially available, but it may be a costly process to extract proteins from pulses. The products may have a bland taste - but they do not have an attractive flavor stimulating the appetite. WO 2006/102907 A1 discloses a fermented protein product including protein derived from yeast and proteinaceous pulse sources with abovementioned functionalities, and at the same time with attractive flavors stimulating the appetite, thereby solving many of the above mentioned problems.

US 2006/0073194 A1 discloses a method for increasing the feed intake and weight gain of a ruminant after a post-receiving stressful event comprising of administering a composition comprising saponins.

US 2007/0071849 A1 discloses a method and compositions for increasing feeding and/or production characteristics in animals using a saponin containing composition in combination with a yeast composition. Commonly when additives, such as saponin, derived from plant material are used as feed additive it is referred to as phytogenies or botanicals. The additives may include herbs e.g. oregano, thyme, non-aqueous extracts such as oleoresins, and essential oils. They act as non-antibiotic growth

promoters. The mode of action is not fully understood but in general an increased feed intake is reported, which is believed to be due to enhanced flavor and odor. Phytogenies are commonly known for their flavoring properties but on the other hand they exert a range of biological effects according to their active compounds. However, these commonly used additives from plant materials are relative expensive additives and their extraction is costly, and there is a need for providing protein products having the same good flavor and odor characteristics or better characteristics at reasonable prices. Accordingly, the main object of this invention is to provide alternative protein products based on proteinaceous pulse sources having good flavor and odor characteristics. Another object is to provide alternative protein products containing phytochemicals at reduced overall costs by utilizing waste plant material as the source of phytochemical additives.

Yet another object is to provide alternative protein feed products providing improvement of overall production parameters in livestock's.

Summary of the invention

The present invention relates to a fermented product comprising 15-80 % by weight of protein derived from one or more proteinaceous pulse sources, which product further comprises up to 10% by weight of one or more plant materials comprising one or more phytochemicals in an amount sufficient to induce a content in the final fermented product of said one or more

phytochemicals of at least 10 ppm.

It has surprisingly been found that when a domestic animal is fed with a feed comprising the fermented product of the invention then an improvement of the overall production parameters is obtained. The plant material, which supplies the fermented protein product with a content of phytochemicals, contributes to this effect.

It has also been found that plant waste materials, such as the press cake, meal or pomace from the production of vegetable oils and wine, can supply with a content of phytochemicals. These plant waste materials are produced in large quantities, and sold at low prices. They do not contribute to favorable flavor and odor but contain the phytochemicals that are useful in the products of this invention.

The present invention in another aspect relates to a method for the manufacture of a fermented product of the invention comprising the following steps:

(i) mixing live yeast with one or more disintegrated proteinaceous pulse sources in a ratio of from about 1 :2 to 1 :100;

(ii) fermenting said yeast/pulse source mixture resulting from step (i) under conditions where the water content does not exceed 60%, for 2-12 hours at a temperature of about 25-60°C, under anaerobic conditions;

(iii) incubating the fermented mixture resulting from step (ii) for 0.5-240 minutes at a temperature of about 70-150°C, the incubation time being sufficient to kill all micro organisms and to inactivate remaining anti-nutritional factors, and provided that a short incubation time is used at high incubation temperatures and a long incubation time is used at low incubation temperatures;

(iv) drying the incubated mixture resulting from step (iii) to a water content of not more than 10% by weight to obtain a fermented protein product, wherein

(v) said one or more plant materials comprising phytochemicals are added either to the disintegrated proteinaceous pulse sources before said mixing step (i) or before or after said drying step (iv). In a third aspect the invention relates to the use of the fermented product for improving feed intake and/or weight gain and/or feed utilization in an animal, e.g. a domestic animal.

It is surprising that the addition of a plant material with no flavor enhancing properties is able to increase the overall production parameters by at least

2% or more as illustrated in the examples. Furthermore it is surprising that this increase in functional properties is obtained by incorporating small amounts of selected phytochemicals in an amount down to about 10ppm.

Definitions

In the context of the current invention, the following terms are meant to comprise the following, unless defined elsewhere in the description.

The terms "about", "around", "approximately", or "~" are meant to indicate e.g. the measuring uncertainty commonly experienced in the art, which can be in the order of magnitude of e.g. +/- 1 , 2, 5, 10, 20, or even 50%.

The term "comprising" is to be interpreted as specifying the presence of the stated part(s), step(s), feature(s), composition(s), chemical(s), or

component(s), but does not exclude the presence of one or more additional parts, steps, features, compositions, chemicals or components. E.g., a composition comprising a chemical compound may thus comprise additional chemical compounds, etc. Phytochemicals:

Comprise plant chemicals that are non-nutritive and not required for sustaining life. The plants produce them for protective reasons. As a part of the food they may have protective and preventive effects.

Some of the functional properties are: Antioxidant, hormonal and anti- bacterial action, and stimulation of enzymes. They can also have a physical action as some phytochemicals bind to cell walls thereby preventing the adhesion of pathogens.

The list of phytochemicals is huge, but the most important in this context are the following types: Flavonoids, phenolic acids, phytosterols, saponins and triterpenoids. Proteinaceous materials:

Comprise organic compounds made of amino acids arranged in a linear chain and joined together by a bond called a peptide bond. At a chain length of up to approximately 50 amino acids the compound is called a peptide, at higher molecular weight the organic compound is called a polypeptide or a protein.

Food products:

Comprise dairy products, processed meat products, sweets, desserts, ice cream desserts, canned products, freeze dried meals, dressings, soups, convenience food, bread, cakes, etc.

Feed products:

Comprise ready-to-use food products for animals such as piglets, calves, poultry, furred animals, sheep, cats, dogs, fish and crustaceans, etc.

Overall production parameters:

Comprise the increase in feed intake and weight gain and better food or feed utilization.

Pharmaceutical products:

Comprise products, typically in the form of a tablet or in granulated form, containing one or more biologically active ingredients intended for curing and/or alleviating the symptoms of a disease or a condition. Pharmaceutical products furthermore comprise pharmaceutically acceptable excipients and/or carriers. The products herein disclosed are very well suited for use as a pharmaceutically acceptable ingredient in a tablet or granulate.

Cosmetic products:

Comprise products intended for personal hygiene as well as improved appearance such as conditioners, lotions and bath formulations. The products herein disclosed are very well suited for use as a cosmetic acceptable ingredient in powdery formulations e.g. bath tub formulations.

Detailed description of the invention In one embodiment of the invention said one or more plant materials are selected from one of more of fruits, nuts, herbs beans and seeds, which may be disintegrated.

The amount of added plant material is up to 10% by weight of the total product which will provide a therapeutic amount of phytochemicals sufficient to increase the functional properties of the final product, e.g. resistance towards oxidation, nitrogen assimilation, wetting and dispersing behavior in watery systems; and improvement of the overall production parameters when used in food or feed vs. the product without the phytochemical fortification.

Accordingly, in another embodiment the one or more phytochemicals are providing the following activities: antioxidant, antibacterial, hormonal, enzyme stimulating and/or improve nitrogen assimilation, wetting and/or dispersing behavior in watery systems.

In another embodiment, the one or more plant materials are waste plant material.

The one or more proteinaceous pulse sources and/or the one or more plant materials may be disintegrated by milling or flaking and/or pressed or solvent extracted in order to reduce the oil content.

The content of phytochemicals in the plant material may in one embodiment have been increased by extraction in a solvent. The plant material has a content of phytochemicals, and the most important types in this context are the following: Flavonoids, phenolic acids,

phytosterols, saponins and triterpenoids. Accordingly, in one embodiment the one or more plant materials comprise one or more of flavonoids, phenolic acids, phytosterols, saponins and triterpenoids.

In another embodiment the one or more plant materials are selected from the press cake, meal or pomace from the production of tea seed (Camellia oleifera) oil, shea nut (Butyrospermum parkii) butter, olive (Olea europaea) oil and grape seed (Vitis vinifera) oil and grape waste material from wine production. These plant waste materials are produced in large quantities, and sold at low prices. They do not contribute to favorable flavor and odor but contain the phytochemicals that are preferred in the products of this invention.

For instance, the saponin contained in the waste materials poses antiprotozoal and anti-bacterial action and the antioxidants in the waste material protects the cells against the action of free radicals and lipid oxidation of fat in the protein product.

In one embodiment of the invention it relates to fermented products

comprising yeast protein that is derived from Saccharomyces cerevesiae or spent brewer's yeast and is present in an amount of 0.3 to 28 % by total weight of the fermented product.

The main reason for using a fermented protein product is to reduce the content of oligosaccharides such as e.g. a-1 ,6-galactosides, and decrease the off-flavors in the proteinaceous pulse source In another embodiment the proteinaceous pulse sources is soybean. The product of the invention may further comprise one or more edible ingredients e.g. emulsifiers, antioxidants or salts.

In one embodiment the content in the final fermented product of the one or more phytochemicals is at least 100 ppm; it may also be at least 200 ppm, at least 500 ppm or at least 1000 ppm.

The fermented product of the invention may be prepared by the method described in WO 2006/102907 A1 , the content of which is incorporated herein. In this method the one or more plant materials comprising phytochemicals may be added to the proteinaceous pulse sources before mixing with the yeast and fermenting or it may be added before or after the final drying of the fermented product. The method of the invention may in one embodiment include after step (ii) a step of (vi) fermenting the yeast/pulse mixture resulting from step (ii) under conditions where the water content does not exceed 60%, for 1 -12 hours at a temperature of about 25-60°C, under aerobic conditions. This method is also conducted as described in WO 2006/102907 A1 .

The present invention also relates to the use of a fermented product of the invention in a food product for human consumption or feed product for animal consumption, and as an ingredient in a cosmetic or pharmaceutical product or a nutritional supplement.

Finally, the invention relates to a protein product comprising 16-79 % by weight of protein derived from one or more proteinaceous pulse sources and up to 10% by weight of one or more plant materials comprising one or more phytochemicals for use in a product of the invention. Examples

Example 1

1 .1 Fermented proteinaceous pulse product based on defatted soybeans

A fermented product was produced as described in WO 2006/102907 A1 . After fermentation and drying the product had the following analysis:

Crude Protein (N x 6,25) 57.1 %

Moisture 6.3 %

Crude fat 2.5 %

Crude fibre 3.5 %

pH (1 :10 aqueous dispersion) 6.3 Furthermore, anti-nutritional factors were found to be the following:

Oligosaccharides 0.5 %

Trypsin Inhibitor 2 600 TlU/g

β-conglycinin 3 ppm

Lectins 1 ppm

Phytic acid-bound phosphorus 0.10 %

1 .2 Plant material comprising saponins from Camellia oleifera

The plant material was formulated as a premix consisting of:

• 10.5% by weight of dried and milled tea (Camellia oleifera) seed meal

• 89.5% by weight of sepiolit powder

1 .3 Plant material comprising antioxidant activity from Vitis vinifera and Olea europaea The plant material was formulated as a premix consisting of:

• 0.8% by weight of grape (Vitis vinifera) press cake

• 0.8% by weight of olive (Olea europaea) press cake

• 98.4% by weight of sepiolit powder

Example 2

Combination of a fermented proteinaceous pulse product and plant material comprising saponins in animal feed.

A fermented proteinaceous pulse product prepared according to Example 1 .1 was used in a 3% addition to a normal piglet feed as a control group.

Performance was compared with a similar feed further comprising 2000ppm, based on the total feed product, of the premix in Example 1 .2.

The trial was carried out with 68 piglets that were weaned at the age of 30 days, average weight of about 7 kg. The piglets were divided into 2 groups at random, similar body weight, genetic base, male and female. The experiment was run in approximately 14 days, during which weight gain and feed intake were recorded.

The mean values for the trial results are shown in the table below:

Control Experiment Index

Start weight 6.7 6.8 -

End weight 9.7 10.1 -

Average daily weight gain 21 1 231 1 10

Average daily feed intake 336 338 101

Kg feed/kg gain 1 .60 1 .46 91 From the results it can be seen that a combination of a fermented product based on soybeans and plant material comprising saponins shows the following improvements:

· 10% on average daily weight gain

• 1 % on average daily feed intake

• 9% better feed utilization

Example 3

Combination of a fermented proteinaceous pulse product and plant material comprising natural antioxidants in animal feed.

A fermented proteinaceous pulse product prepared according to Example 1 .1 was used in a 3% addition to a normal piglet feed as a control group.

Performance was compared with a similar feed further comprising 2000ppm, based on the total feed product, of the premix in Example 1 .3.

The trial was carried out with 68 piglets that were weaned at the age of 30 days, average weight of about 7 kg. The piglets were divided into 2 groups at random, similar body weight, genetic base, male and female. The experiment was run in approximately 14 days, during which weight gain and feed intake were recorded.

The mean values for trial results are shown in the table below: Control Experiment Index

Start weight 6.7 6.8 -

End weight 9.7 9.8 -

Average daily weight gain 21 1 218 103

Average daily feed intake 336 340 102

Kg feed/kg gain 1 .59 1 .56 98

From the results it can be seen that a combination of a fermented product based on soybeans and plant material comprising natural antioxidants shows the following improvements:

· 3% on average daily weight gain

• 2% on average daily feed intake

• 2% better feed utilization

Example 4

Fermented protein product based on defatted soybeans comprising saponins from Camellia oleifera

A fermented protein product based on defatted soybeans was produced as described in Example 1 .1 . After drying and milling it was mixed with 0.8% by weight of dried and milled tea (Camellia oleifera) seed meal (a similar commercial available product is obtainable with the trade name Nor-Spice TSC Powder from NOR-FEED A S, Denmark).

The final product had the following composition:

Protein (Nx6.25) 53.0%

Carbohydrates 25.2%

Water 8.0%

Ash 6.0%

Crude fiber 5.0% Fat 2.0%

Plant elements from

Camellia oleifera 0.8% Further typical data: pH (10% suspension) 6.4

Particle size (< 200μ) 70%

Wetting properties Excellent

Water binding 1 : 5

The product is suited as an ingredient for production of feed and specifically for application in feed for piglets.

The product is characterized by a low content of anti-nutritional matter and improving the overall production parameters, as illustrated in Example 2.

Summary Listing of References WO 2006/102907 A1 US 2006/0073194 A1 US 2007/0071849 A1