Feed or food ingredient derived from the fiber-rich biomass of soybean husks

Abstract

The present invention provides a feed or food ingredient derived from soybean hulls, which comprises dietary fiber derived from soybean hulls in the form of soluble and insoluble polysaccharides, wherein the dietary fiber derived from soybean hulls has been partially degraded by one or more carbohydrases selected from mannanase, pectinase, xylanase, glucanase, and cellulase into oligosaccharides having 3 to 30 monomer sugar units (oligosaccharide DP3-30), and the feed or food product contains 5% by mass or less of monosaccharides, and preferably provides a prebiotic effect.

Description

[Technical field]

[0001] FIELD OF THE PRESENT APPLICATION The present invention relates to a feed or food ingredient derived from the fibre-rich plant biomass of soybean hulls, to a method for producing said feed / food ingredient, to uses of said feed / food ingredient and to feed or food products comprising said feed / food ingredient. [Background technology]

[0002] 2. Background of the Invention Fiber is becoming increasingly important in alternative feeding of production animals. Previously, fiber was considered an antinutritional factor that reduces the energy concentration and digestibility of the feed, leading to reduced animal performance. However, in the last 20 to 30 years, increasing evidence has shown that the inclusion of certain fibers is beneficial for animal performance and the development of a healthy intestinal tract; Agyekum & Nyachoti (2017), Molist et al. (2009). Both the content of soluble and insoluble non-starch polysaccharides (NSPs) and the content of prebiotic oligosaccharides are part of the dietary fiber that can have a positive effect in animals. More generally, fiber is currently being intensively investigated as one of the solutions to the limited use of antibiotics and zinc in the animal sector. The reduced use of antibiotics and zinc leads to an increase in poor performance, disease, and diarrhea, which is a serious problem.

[0003] Dedicated fiber sources are increasingly being used in feeding monogastric animals. Soybean hulls are such dedicated fiber sources with high total and insoluble fiber content. Surprisingly, the present inventors have now found that when soybean hulls, a fiber biomass, are treated with specific carbohydrate targeting enzymes, these fibers can release significantly more oligosaccharides than many other natural biomasses with high fiber content. Depending on the type of targeting enzyme applied, the nature of the oligosaccharides released from soybean hulls and other biomasses can be different.

[0004] In the feeding of some animals, it is now common to add "exogenous" enzymes to the final feed formulation, but the enzymes do not act before being ingested with the rest of the diet, but in the animal's gut environment (Kiarie et al. 2013, Scapini et al. 2018). The use of carbohydrase enzymes as production aids to increase the prebiotic oligosaccharide content of feed biomass has only been described to a limited extent. The potential effect of some oligosaccharides is expected to be prebiotic when oligosaccharide-rich sources are fed to production animals, such as pigs and poultry, resulting in improved animal performance and health. For human food products or research purposes, biomass is treated with enzymes and / or physical / chemical means to produce, extract and purify prebiotic oligosaccharides. Alternatively, some prebiotic products are chemically synthesized and purified for the same use (e.g. polydextrose, dextrin). This type of product is expensive and therefore not used to a significant extent in animal feed; Babber et al. (2015); Aachary & Prapulla (2011); Dotsenko et al. (2017); Kurakake et al. (2006).

[0005] At present, there does not appear to be any feed or food product on the market that originates from soybean hulls and has increased specific oligosaccharide content compared to untreated soybean hulls due to carbohydrase enzyme treatment.There is also no prior art that describes carbohydrase enzyme-treated soybean hulls for consumption; some documents only describe that soybean hulls can be added to the diet together with an external carbohydrase enzyme source before feeding to animals.

[0006] The inventors have tested different soy hull products treated with carbohydrate targeting enzymes such as mannanase, xylanase, or pectinase in piglet feeding trials and in vitro piglet trials and found improved performance compared to controls. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Maribo (2005): "Tilsetningsstoffer til svin. Landsudvalget for svin" Summary of the Invention [Problem to be solved by the invention]

[0008] The object of the present invention is to provide a new feed or food ingredient with a unique composition originating from the fiber-rich plant biomass of soybean hulls.

[0009] Another object is to provide new and improved feed or food ingredients originating from soybean hulls, in a cost manner that is favorable for the applied carbohydrate-targeting enzymes, the source of which contains both a certain amount of complex carbohydrates in the form of dietary fiber and an increased amount of prebiotic oligosaccharides obtained by the action of carbohydrate-targeting enzymes, which is advantageous when used in animal or human diets.

[0010] Another object is to provide a new feed or food ingredient obtainable by treating soybean hulls with carbohydrate-targeting enzymes.

[0011] It is a further object to provide improved feed or food ingredients containing prebiotic oligosaccharides, which have increased digestibility compared to untreated (raw) soybean hulls.

[0012] Finally, it is an objective to provide new products that have high water binding capacity and low viscosity; both of these properties are important for the application of the product in animal diets.

[0013] These objectives are met by the products of the present invention. [Means for solving the problem]

[0014] SUMMARY OF THE PRESENT APPLICATION Thus, in a first aspect, the present invention relates to a feed or food ingredient derived from soybean hulls, comprising dietary fibre derived from soybean hulls in the form of soluble and insoluble polysaccharides, wherein the dietary fibre derived from soybean hulls has been partially degraded by one or more carbohydrases selected from mannanase, pectinase, xylanase, glucanase and cellulase into oligosaccharides having 3 to 30 monomeric sugar units (oligosaccharide DP3-30), and wherein the feed or food product comprises not more than 5% by weight of monosaccharides.

[0015] In the context of the present invention, oligosaccharides DP3-30 derived from dietary fiber do not include raffinose, stachyose, and verbascose.

[0016] Surprisingly, the inventors found that when various different fiber-rich plant biomasses were treated with selected carbohydrases, i.e., enzymes acting on complex polysaccharides, soybean hulls stood out as a biomass capable of releasing extremely large amounts of oligosaccharides. Also, several selected mannanases, pectinases, xylanases, cellulases, and β-glucanases produced significant amounts of soybean hull-derived oligosaccharides compared to those typically found from other biomasses. It is expected that some of these oligosaccharides may act as prebiotics in animal feed or human food, depending on the chemical properties of the oligosaccharides.

[0017] It is even more surprising that such products, when applied in animal diets, can improve animal performance, for example piglet and poultry performance. The inventors have confirmed this in piglet feeding trials. The known mechanisms of action behind the improvement of performance when using prebiotic oligosaccharides are improvement / stabilization of healthy microbial community in the gut, reduction of risk of intestinal infection and inflammation, improvement of gut function (stool bulk, regularity and consistency), improvement of gut barrier function, regulation / modulation of immune function and modulation of gastrointestinal peptide production and energy metabolism; Roberfroid et al. (2010). The best known and most thoroughly studied and documented oligosaccharides are fructooligosaccharides (FOS), typically derived from inulin; Ghoddusi et al. (2007); Probert et al. (2004); and mannan oligosaccharides (MOS); Corrigan et al. (2015); Zivkovic et al. (2011); Kim et al. (2010). In the case of MOS, the types of MOS whose effects are well documented are those derived from yeast cell walls. These are the α-1,3 and α-1,6 branched mannans. Examples of the emerging group of prebiotic oligosaccharides are xylooligosaccharides (XOS); Liu et al. (2018); Moura et al. (2008); Aachary & Prapulla (2011); Dotsenko et al. (2017); Nielsen et al. (2014); pectin-derived oligosaccharides (POS); Babber et al. (2015); Chung et al. (2017); Strube et al. (2015); and β-glucan-derived oligosaccharides; Meyer et al. (2015); Miguez et al. (2016). Galactooligosaccharides (GOS) can be synthesized from lactose, which is also a new prebiotic oligosaccharide; Torres et al. (2010). The chemical properties that are important for whether an oligosaccharide has a prebiotic effect are the type of chemical bond between the sugar moieties, the identity of the sugar molecule, and its branching; Markowiak & Slizewska (2018); Pourabeden & Zhao (2015), Kim et al. (2019).

[0018] The recommended amount of MOS in animal feed for pigs is approximately 0.25 to 0.5% of the final feed formulation and the recommended amount of FOS is approximately 0.03 to 1.25% of the feed formulation. Maribo (2005): "Tilsetningsstoffer til svin. Landsudvalget for svin". This supports the idea that even small amounts of prebiotic oligosaccharides added to production animal feed can make a difference.

[0019] In a second aspect, the present invention provides a method for producing a feed or food ingredient according to the invention, comprising the steps of: mixing the soybean hulls with one or more carbohydrases selected from mannanase, pectinase, xylanase, glucanase, and cellulase; - hydrolyzing the mixture at a temperature of 20 to 60°C for a period of 1 to 48 hours under conditions of a dry matter (DM) content of not more than 55% by weight; Optionally, completely inactivating the activity of one or more carbohydrases; - isolating the feed or food ingredient; The present invention relates to a method comprising the steps of:

[0020] The present invention further provides a feed or food product or dietary supplement for production animals containing 0.5 to 99% by weight of the feed or food ingredient of the present invention, such as a feed product for use in diets for production animals in need of prebiotic oligosaccharides, preferably newborn and young animals, such as piglets, calves, and poultry.

[0021] definition In the context of the present invention, the following terms, unless defined elsewhere herein, are intended to include the following:

[0022] The term "comprising" should be interpreted as specifying the presence of stated moieties, steps, features, compositions, chemicals, or ingredients, but not excluding the presence of one or more additional moieties, steps, features, compositions, chemicals, or ingredients. Thus, for example, a composition that includes a compound can include additional compounds, etc.

[0023] biomass: It includes biological materials produced by photosynthesis and that can be used in industrial production. In the context of the present invention, biomass refers to fiber-rich plants in the form of soybean hulls. Soybean hulls can be applied in raw form or in a disintegrated and / or pretreated form, depending on the nature and the approach of the person skilled in the art to the particular raw material.

[0024] Soy products: Soybean products, in the context of the present invention, in particular products derived from soybean husks, and plants in the form of mixtures thereof. Soybeans may be from any soybean source, for example from South or North America or Asia or Europe, and may be of genetically modified origin (GMO) or of non-genetically modified origin (non-GMO).

[0025] Soy hulls Soybean hulls are a dedicated fiber source with high total and insoluble fiber content and also contain some protein. Soybean hulls are a by-product of soybean processing and consist of the coating of soybeans. Soybean hulls contain complex carbohydrates such as pectin, hemicellulose, and cellulose and are a good source of dietary fiber.

[0026] Disintegrated: For example, it means that it is disintegrated by chemical or physical means according to methods known in the art, such as milling, blending, cooking, and / or acid or alkali treatment. The skilled person knows whether disintegration is necessary and, if so, which disintegration may be suitable for the particular plant biomass applied in the present invention.

[0027] Oligosaccharides: Oligosaccharides are typically defined as saccharide oligomers containing a small number (3-10) of component monomeric sugars. In the context of the present invention, oligosaccharides are more broadly defined and can be saccharide oligomers containing from 3 to 30 monomeric sugar units.

[0028] Oligosaccharides with 3 to 30 sugar units / Oligosaccharide DP3~30 In the context of the present invention, these terms refer to saccharide oligomers containing 3-30 monomeric sugar units, a feature that applies in the definition of the new products of the present invention. In the context of the present invention, soybean oligosaccharides contained in raw soybean hulls, in particular raffinose, stachyose and verbascose, are not included in DP3-30 as defined herein.

[0029] Prebiotic Oligosaccharides Oligosaccharides considered prebiotics must not be hydrolyzed or absorbed in the upper gastrointestinal tract, but must be selectively assimilated by beneficial microorganisms in the colon to promote beneficial luminal or systemic effects; Meyer et al. (2015); Miguez et al. (2016). They can be fermented by microorganisms in the hindgut, but must "survive" until they get there; Smiricky-Tjardes et al. (2003).

[0030] Monosaccharides / Simple sugars Monosaccharides, or simple sugars, are simple sugar monomers containing 5 and / or 6 carbon atoms and are the basic units of sugars and carbohydrates. In the context of the present invention, monosaccharides / simple sugars are in particular glucose, fructose and galactose.

[0031] polysaccharide Polysaccharides are sugar polymers that contain multiple component monomeric sugars, also known as complex carbohydrates. Polysaccharides can be soluble and insoluble.

[0032] Fiber Dietary fiber includes soluble and insoluble non-starch polysaccharides and may include oligosaccharides, lignin, and resistant starch. The raw biomass selected in the context of the present invention contains only trace amounts of oligosaccharides and resistant starch, or is substantially free of resistant starch.

[0033] Carbohydrase A carbohydrase in the context of the present invention is any enzyme capable of hydrolyzing any carbohydrate structure, such as mannanases, pectinases, xylanases, glucanases, and cellulases.

[0034] yeast In the context of the present invention, the yeast may in particular be chosen from among Saccharomyces cerevisiae strains, such as spent brewer's yeast and spent distiller's yeast and spent yeast from wine production, bioethanol yeast or spent yeast from bioethanol production, baker's yeast, and yeast strains which ferment C5 sugars.

[0035] microorganisms Microorganisms are organisms that are microscopic and therefore too small to be seen by the naked human eye. Microorganisms include bacteria, fungi, archaea, protists, and viruses.

[0036] lactic acid bacteria They are commonly found in decaying plants and dairy products and produce lactic acid as the main metabolic end product of carbohydrate bioconversion. Lactic acid bacteria are a genus of microorganisms that produce organic acids, such as lactic acid and some acetic acid, as metabolic products of carbohydrate bioconversion. The genus includes, but is not limited to, Lactobacillus, Pediococcus, Lactococcus, Enterococcus, Weisella, Streptococcus, and Leuconostoc.

[0037] Other genera In the context of the present invention, other genera refer to other bacterial genera that are most relevant in the context of the present invention. These include several genera that also produce organic acids, such as lactic acid and acetic acid, as metabolic products of carbohydrate bioconversion, but often to a lesser extent than lactic acid bacteria. In the context of the present invention, other genera other than lactic acid include, but are not limited to, Bacillus, Bifidobacterium, Brevibacillus, Propionibacterium, Clostridium, and Geobacillus. Certain strains are used as probiotics.

[0038] feed products This includes ready-to-use feed or feed ingredients for production animals such as piglets, calves, poultry, fur animals, and sheep.

[0039] food products Includes ready-to-use foods or food ingredients for human nutrition.

[0040] The invention is illustrated in the following drawings. [Brief description of the drawings]

[0041] [Figure 1] Figure 1 shows initial screening results after incubation of soybean hull biomass with different carbohydrate-targeting enzymes under pH control. The measured oligosaccharides DP3-30 do not contain raffinose, stachyose, or verbascose. [Diagram 2] Figure 1 shows a similar screen after similar incubation of soybean hull biomass without pH adjustment, but with the addition of baker's yeast. The measured oligosaccharides DP3-30 do not contain raffinose, stachyose, or verbascose. [Diagram 3] FIG. 1 shows TLC results of soybean hulls incubated with different carbohydrase enzymes. [Figure 4]Figure 1 shows the effect of hydrolysis time when Vland pectinase or mannanase was used to incubate soybean hull biomass. The measured oligosaccharides DP3-30 do not contain raffinose, stachyose, or verbascose. [Diagram 5] Figure 1 shows the effect of hydrolysis time and enzyme dosage when soybean hull biomass is incubated with different carbohydrase enzymes. The measured oligosaccharides DP3-30 do not contain raffinose, stachyose, and verbascose. [Figure 6] As a comparison, FIG. 1 shows the quantification of oligosaccharides DP3-30 (not including raffinose, stachyose, and verbascose) in different biomasses, i.e., either raw biomass, biomass incubated without enzyme, or enzyme-treated biomass after incubation with carbohydrase enzyme. [Figure 7] FIG. 1 shows TLC of soybean hull biomass after incubation with Depol 793L (pectinase), Strowin xylanase, mannanase, and cellulase, respectively. [Figure 8] FIG. 1 shows the water retention in soybean hulls compared to different biomasses, either in the raw form or after treatment according to the present invention. [Figure 9] FIG. 1 shows the viscosity of different biomasses, either in the raw form or after treatment according to the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0042] Detailed Description of the Invention In a first aspect, the product of the invention comprises: In a first aspect, the present invention relates to a feed or food ingredient derived from soybean hulls, comprising dietary fibre derived from soybean hulls in the form of soluble and insoluble polysaccharides, wherein the dietary fibre derived from soybean hulls has been partially degraded by one or more carbohydrases selected from mannanase, pectinase, xylanase, glucanase and cellulase into oligosaccharides having from 3 to 30 monomeric sugar units (oligosaccharide DP3-30), and wherein the feed or food product comprises up to 5% by weight of monosaccharides.

[0043] In this context, oligosaccharides DP3-30 derived from dietary fiber do not include raffinose, stachyose and verbascose.

[0044] In one embodiment of this aspect, the dietary fiber from soybean hulls is partially degraded by one or more carbohydrases into oligosaccharides, providing a prebiotic effect.

[0045] In any embodiment, the feed or food product contains 5% or less by weight of monosaccharides; for example 4% or less; 3% or less; 2% or less; 1% or less; 0.5% or less by weight; or is substantially free of monosaccharides or is free of monosaccharides.

[0046] In another embodiment, the feed or food ingredient may comprise 4% or more by weight of oligosaccharides having 3 to 30 monomeric sugar units (oligosaccharides DP3-30), such as 5% or more, 6% or more, 7% or more, 8% or more, 9% or more, 10% or more, 12% or more, 15% or more, or 20% or more by weight of oligosaccharides having 3 to 30 monomeric sugar units (DP3-30). In this context, oligosaccharides DP3-30 derived from dietary fibre exclude raffinose, stachyose, and verbascose.

[0047] In any of the embodiments of the invention in the first aspect, the feed or food ingredient may optionally further comprise live yeast, such as a live yeast selected from among Saccharomyces cerevisiae strains including spent brewer's yeast, baker's yeast, spent distiller's yeast, and spent yeast from wine production, bioethanol yeast, or spent yeast from bioethanol production, and yeast strains that ferment C5 sugars. The live yeast may for example be present in an amount of 0.05 to 10%, such as 0.10%, 0.15%, 0.20%, 0.25%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5, 5%, 6%, 7%, 8% or 9%.

[0048] In any of the embodiments of the invention in the first aspect, the oligosaccharides raffinose, stachyose and / or verbascose present in raw soybean hulls may optionally be fully or partially degraded.

[0049] In any of the embodiments of the invention in the first aspect, the feed or food ingredient may optionally further comprise one or more microorganisms and / or metabolic products of carbohydrate degradation of soybean hulls by one or more microorganisms.

[0050] In any of the embodiments of the invention in the first aspect, the feed or food ingredient may optionally further comprise metabolic products of carbohydrate degradation of soybean hulls by one or more microorganisms selected from lactic acid bacteria and / or lactic acid bacteria, such as Lactobacillus, Pediococcus, Lactococcus, Enterococcus, Weissella, Streptococcus, and Leuconostoc.

[0051] In any of the embodiments of the invention in the first aspect, the feed or food ingredient may optionally further comprise metabolic products of carbohydrate degradation of soybean hulls by one or more microorganisms and / or one or more microorganisms selected from the genera Bacillus, Bifidobacterium, Brevibacillus, Propionibacterium, Clostridium, and Geobacillus.

[0052] In any of the embodiments of the invention in the first aspect, the feed or food ingredient may be derived from soybean hulls, where the soybean hull fibre has been degraded by one or more β-mannanases to oligosaccharides comprising 4% or more oligosaccharides having 3 to 30 monomeric sugar units, such as 5% or more, 10% or more, 15% or more, or 20% or more oligosaccharides having 3 to 30 monomeric sugar units (DP3-30), by weight. In this context, oligosaccharides DP3-30 derived from dietary fibre do not include raffinose, stachyose, and verbascose.

[0053] In any of the embodiments of the invention in the first aspect, the feed or food ingredient may be derived from soybean hulls, where the soybean hull fibre has been degraded by one or more pectinases to oligosaccharides comprising 4% or more by weight of oligosaccharides having 3 to 30 monomeric sugar units, such as 5% or more, 10% or more, 15% or more, or 20% or more by weight of oligosaccharides having 3 to 30 monomeric sugar units (DP3-30). In this context, oligosaccharides DP3-30 derived from dietary fibre do not include raffinose, stachyose, and verbascose.

[0054] In any of the embodiments of the invention in the first aspect, the feed or food ingredient may be derived from soybean hulls, where the soybean hull fibre has been degraded by one or more xylanases to oligosaccharides comprising 4% or more by weight of oligosaccharides having 3 to 30 monomeric sugar units, such as 5% or more, or 10% or more by weight of oligosaccharides having 3 to 30 monomeric sugar units (DP3-30). In this context, oligosaccharides DP3-30 derived from dietary fibre do not include raffinose, stachyose and verbascose.

[0055] In any of the embodiments of the invention in the first aspect, the feed or food ingredient may be derived from soybean hulls, where the soybean hull fibre has been degraded by one or more glucanases to oligosaccharides comprising 4% or more by weight of oligosaccharides having 3 to 30 monomeric sugar units, such as 5% or more, or 10% or more by weight of oligosaccharides having 3 to 30 monomeric sugar units (DP3-30). In this context, oligosaccharides DP3-30 derived from dietary fibre do not include raffinose, stachyose and verbascose.

[0056] In any of the embodiments of the invention in the first aspect, the feed or food ingredient may be derived from soybean hulls, where the soybean hull fibre has been degraded by several carbohydrases into oligosaccharides having 3 to 30 monomeric sugar units (DP3-30).

[0057] In a second aspect, the method of the present invention comprises: In a second aspect of the invention, the present invention relates to a method for producing a feed or food ingredient according to the invention, comprising the steps of: mixing the soybean hulls with one or more carbohydrases selected from mannanase, pectinase, xylanase, glucanase, and cellulase; - hydrolyzing the mixture at a temperature of from 20 to 60°C for a period of from 1 to 48 hours under conditions in which the dry matter (DM) content is not more than 55% by weight, Optionally, completely inactivating the activity of one or more carbohydrases, Processes for isolating feed or food ingredients The present invention relates to a method comprising the steps of:

[0058] Those skilled in the art will be able to select a suitable amount of carbohydrase enzyme to meet the requirements of the process depending on the process conditions, intended results, and optimal process economics.

[0059] In one embodiment of this aspect, the fiber-rich plant biomass may be disintegrated before being mixed with one or more carbohydrases. The means for disintegration may be physical or chemical, and the skilled person knows whether disintegration is appropriate and necessary and which means are applicable to a particular biomass.

[0060] In any embodiment of the invention in the second aspect, the method comprises a step of completely inactivating the activity of one or more carbohydrases, carried out after the step of hydrolyzing the mixture, which ensures that the oligosaccharides DP3-30 are not degraded to monosaccharides.

[0061] In any embodiment of the invention in the second aspect, the dry matter content is 55% by weight or less, such as 53% by weight or less, 51% by weight or less, 50% by weight or less, 48% by weight or less, 46% by weight or less, or 45% by weight or less.

[0062] In any embodiment of the invention in the second aspect, the reaction time is from 1 to 48 hours, for example 2, 4, 6, 8, 12, 16, 20, 24, 28, 32, 36, 40 or 44 hours.

[0063] In any embodiment of the invention in the second aspect, the reaction temperature is from 20 to 60°C, for example 25, 30, 32, 35, 37, 40, 45, 50, 55, or 58°C.

[0064] In any embodiment of the invention in the second aspect, the skilled artisan can select and adapt the reaction time and reaction temperature in proportion to the selected carbohydrase and its amount.

[0065] In any embodiment of the invention in the second aspect, before the hydrolysis step, live yeast, for example selected from among Saccharomyces cerevisiae strains including spent brewer's yeast, baker's yeast, spent distiller's yeast, and spent yeast from wine production, bioethanol yeast, or spent yeast from bioethanol production, and yeast strains fermenting C5 sugars, may be added to the mixture of soybean hulls and carbohydrase in an amount of, for example, 0.05 to 10%, for example 0.1%, 0.15%, 0.20%, 0.25%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5, 5%, 6%, 7%, 8% or 9%. The skilled person can select a suitable amount depending on the process conditions and the intended result.

[0066] In any embodiment of the present invention in the second aspect, one or more microorganisms may be added to the mixture of soybean husk and carbohydrase before the hydrolysis step. The addition of microorganisms is suitable for the conversion of carbohydrates present in soybean husk into useful metabolic products, especially organic acids such as lactic acid and acetic acid. Those skilled in the art can select the suitable amount depending on the process conditions and the intended results.

[0067] In any embodiment of the present invention in the second aspect, lactic acid bacteria, such as Lactobacillus, Pediococcus, Lactococcus, Enterococcus, Weissella, Streptococcus and Leuconostoc, may be added to the mixture of soybean husk and carbohydrase before hydrolysis.The addition of lactic acid bacteria is suitable for converting the carbohydrates present in plant biomass into useful metabolic products, especially organic acids, such as lactic acid and acetic acid.Those skilled in the art can select suitable amounts according to process conditions and intended results.

[0068] In any embodiment of the present invention in the second aspect, one or more microorganisms selected from the genera Bacillus, Bifidobacterium, Brevibacillus, Propionibacterium, Clostridium, and Geobacillus may be added to the mixture of soybean hulls and carbohydrase before the hydrolysis step. Those skilled in the art can select a suitable amount depending on the process conditions and the intended results.

[0069] In any embodiment of the present invention in the second aspect, α-galactosidase may be added to the mixture of soybean hulls and carbohydrase before the hydrolysis step. The addition of α-galactosidase is suitable for decomposing raffinose, stachyose, and verbascose present in soybean hulls. Those skilled in the art can select a suitable amount depending on the process conditions and the intended results.

[0070] In a third aspect, a feed or food product or a dietary supplement of the invention comprising: In a third aspect the invention relates to a feed or food product or a dietary supplement containing 0.5 to 99% by weight of the feed or food ingredient according to the invention.

[0071] In any embodiment of the invention in its third aspect, the feed product or nutritional supplement may be for use in diets for production animals, such as diets to improve performance in production animals, particularly newborn and young animals, such as piglets, calves, and poultry.

[0072] In any embodiment of the invention in the third aspect, the feed product or dietary supplement may be for use in diets for production animals, particularly newborn and young animals, such as piglets, calves, and chickens, that require prebiotic oligosaccharides.

[0073] In a fourth aspect, the present invention relates to the use of a feed ingredient according to the invention in diets for producing animals, in particular newborn and young animals, such as piglets, calves, and chickens.

[0074] Working Example Materials and Methods Enzymes and Enzyme Suppliers: BIO-CAT, Vland Biotech Inc, Strowin, Jinan BestZyme, Winovazyme, Habio, Challenge Group, Biocatalysts Ltd and DSM. The enzymes were supplied between April 2018 and December 2019, and the specific names of the enzymes are shown below.

[0075] [Table 1]

[0076] Thin layer chromatography (TLC method) A 10% DM suspension of the sample was first prepared and homogenized for 30 seconds using an Ultra Thorax instrument at level 2. The sample was stirred for 30 minutes at room temperature and centrifuged at 3000 G for 10 minutes. The supernatant was used directly for TLC.

[0077] 1.4 μl of sample and different standard mixes were added to the TLC plate. After drying, the plate was placed in a chromatography tank for 50-60 min, followed by drying and immersion in developer solution for 2 s. After drying, the TLC pattern was visible after a short time at 150 °C. The chromatography fluid consisted of demineralized water containing 46 vol / vol% n-butanol and 31 vol / vol% pyridine. The developer solution consisted of 2 g diphenylamine, 84 ml acetone, 2 ml aniline and 15 ml 85% phosphoric acid per 100 ml.

[0078] Ion exchange chromatography analysis (Dionex) A 10% DM suspension of the samples was prepared and homogenized for 30 seconds using an Ultra Turrax T25 instrument, IKA, at level 2. Samples were stirred for 30 minutes at room temperature and centrifuged at 3000 G for 10 minutes. The supernatant was used for Dionex or Viscotek analysis.

[0079] Oligosaccharides and sugars were separated and quantified by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD, Dionex). Oligosaccharides were estimated using an external maltodextrin standard. Samples were injected onto a CarboPacPA-200 column using a flow rate of 0.4 mL min-1, isocratic NaOH at 150 mM and the following NaOAc gradient profile: 0-5 min: linear gradient from 0 to 110 mM, 5-30 min: convex gradient from 110 to 350 mM.

[0080] Size exclusion chromatography analysis (Viscotek) Samples were examined by size exclusion chromatography (SEC) using a Viscotek system (Malvern, UK) equipped with two columns in series: GS-320 HQ and GS-620 HQ columns (Shodex) mounted on a TDA 302 module (triple detector array) containing a refractive index detector (RI), a four-bridge viscometer detector (VIS) and a light scattering detector (LS). LS included right angle light scattering (RALS) and low angle light scattering (LALS) measuring scattered light at 7° and 90° to the incident beam. The instrument was calibrated using pullulan (50 kDa, polydispersity 1.07, Showa Denko) solubilized in MilliQ water (1 mg / mL) at 1000 rpm for 120 min at 99°C. Elution was performed with 50 mM ammonium formate (HCO2NH4) buffer and a flow rate of 0.5 mL min-1. Samples were filtered through a 0.22 μm centrifugal filter and 50 μl of sample was injected onto the column (GPCmax module) and separated at 60° C. Authentic and well-characterized linear maltooligosaccharides were used as external standards. Data were analyzed using OmniSec software 4.7 (Malvern Instrument, Ltd.).

[0081] Example 1 A small-scale screen demonstrating the effect of carbohydrases in soybean hulls. A total of 40 commercially available carbohydrase enzymes covering at least the common groups of mannanases, xylanases, cellulases, β-glucanases, and pectinases from different suppliers were tested in soybean hull biomass. Enzymes were dosed according to enzyme price, and doses equivalent to a certain price per mass of soybean hull dry matter were added and compared accordingly. Soybean hulls were delivered as dry pills and coarsely ground in a blender. Enzymes and soybean hulls were mixed and incubated at 32°C for 8-44 hours at a dry matter (DM) content of 45-48% by mass. Approximately 90 g of soybean hulls were used in this small-scale screening, and α-galactosidase was added to all samples to remove the oligosaccharides stachyose and raffinose in the soybean hull biomass. In some experiments, baker's yeast was added to the reaction mixture. In some experiments, the pH was adjusted to 4.5 with H2SO4.

[0082] After hydrolysis of the biomass, its oligosaccharide DP3-30 (oligosaccharides with 3-30 sugar units) content was determined by either the Dionex or Viscotek method. A TLC method was also applied for further quantification of the enzymatic cleavage patterns into oligosaccharides by different enzymes.

[0083] Figures 1A, B and C show the initial screening results after pH adjustment, 21 hour incubation and analysis of the products by the Dionex method.

[0084] Figure 2 shows the screening results after 8-44 hours of incubation without pH adjustment and analysis of the products by the Dionex and Viscotek methods. When more analyses were performed with the same enzyme, the average values ​​were used in the figures.

[0085] Figure 3 shows the sugar composition of soybean hulls subjected to treatment with different carbohydrase enzymes, clearly demonstrating that different types of enzymes cleave in soybean hulls in different manners, resulting in patterns typical of mannanase, xylanase, pectinase, and β-glucanase / cellulose, respectively. The reference sugar mixture displayed in lanes 1-4 contains, from top to bottom, the following: Mix 1: Xylose, sucrose, raffinose, stachyose Mix A: arabinobiose, arabinose, mannobiose, mannotriose Mix B: Xylose, xylobiose, xylotriose, mannobiose, mannotriose Mix C: Galactose, Galacturonic Acid

[0086] The average maximum obtained oligosaccharide levels obtained in the small scale screening with the different enzyme panels after 16 hours of hydrolysis are shown as the average assay values ​​for the best enzymes below:

[0087] [Table 2]

[0088] Example 2 Demonstrating the effect of carbohydrases on soybean hulls at pilot scale In a horizontal, slowly rotating reactor, three hydrolysis experiments were carried out with 450 kg of soybean husks and the most effective enzymes from the screening examples, as well as baker's yeast and α-galactosidase, according to the pilot experiment. The soybean husk pills were not pre-disintegrated, the initial DM and temperature were 48% by weight and 30°C, and the incubation time was 16 h.

[0089] A Viscotek analysis was performed. The content of oligosaccharides DP3-30 (oligosaccharides with 3-30 sugar units) was recalculated as a mass % of total soybean hull biomass.

[0090] [Table 3]

[0091] Example 3 The effect of incubation time and enzyme dosage is shown. The effect of incubation time was investigated using either hydrolysis conditions at a pH of 4.5 (Figure 4) or at unadjusted pH (Figure 5) as described in Example 1. Figure 5 also shows the effect of including half the enzyme dosage. As expected, the experiment shows that the effect of incubation time is enzyme dependent.

[0092] Example 4 Showing the effect of oligosaccharide levels and carbohydrases in other biomasses compared to soybean husks Oligosaccharide levels were examined in a variety of different biomasses of different morphologies: a) Raw biomass (control); b) biomass incubated without the addition of carbohydrase enzymes (for biomass containing raffinose, stachyose, and verbascose, α-galactosidase was added to remove these oligosaccharides), or c) Biomass incubated with commercial carbohydrase enzyme.

[0093] Enzymes were dosed according to price and doses equivalent to a fixed price were added per mass of biomass dry matter and compared accordingly. Sugar beet pulp and soybean hull biomass were delivered as dry pellets and were roughly ground in a blender. Other biomasses were used as delivered. Enzymes, baker's yeast and biomass were mixed and incubated at 45 or 48% DM at 32 or 37°C for 16, 20 or 44 hours. After hydrolysis, the biomass was analyzed as described in Example 1. The results are shown in Figure 6. Soybean oligosaccharides (raffinose, stachyose and verbascose) were subtracted if present in the raw biomass.

[0094] FIG. 6 shows the Dionex or Viscotek quantification of oligosaccharides in different biomasses: either raw biomass, biomass incubated without enzyme, or biomass treated with enzyme.

[0095] Comparing the relative (compared to the starting levels in the feedstock) release of oligosaccharides, soybean hulls stand out as the biomass capable of releasing the highest relative amount.

[0096] [Table 4]

[0097] Example 5 Showing the effect of incubating DM on soybean hulls Experiments conducted as described in Example 1 at unadjusted pH were performed to test three starting biomass DM levels: 45, 48 and 51% DM. The results are shown in Figure 7 which shows the TLC and indicates no effect of increasing the DM% from 45% to 51% with Depol 793L (pectinase), Strowin xylanase, mannanase and cellulase.

[0098] Example 6 A piglet feeding trail was carried out at the Danish test station: Skjoldborg teststation, TestGris, SvineRadgivningen, Herning, Denmark, from May to August 2018. Using a conventional pig production system, four diets were tested, including: 1. Control without soybean hulls; 2. FaserGold, extruded soy hulls, amounting to 2.5% by DM weight; 3. Soybean hulls incubated with α-galactosidase and baker's yeast for 8 hours, milled and dried, at 2% by DM weight; 4. Soybean hulls incubated with α-galactosidase, Ronozyme VP, and baker's yeast for 7 hours, milled and dried, 2% by DM weight.

[0099] The study was carried out in three separate phases (A, B, and C) of two weeks each, for six weeks after weaning. The soy hull product was included only during phase A. In phases B and C, the piglets were fed the same diet. None of the diets contained antibiotics or therapeutic levels of veterinary ZnO. All diets were fed ad libitum. The pigs had constant access to fresh water. The diets in phase A were based on wheat (35-38%) and barley (15%), with Hamlet Protein's HP300 (19%) used as the protein source. Equal amounts of "premix" were used in each diet (25%), and soybean oil was used to equalize the energy levels in the diets (2.7-3.7%). The wheat and barley were milled and the diets pelleted. The diets in phases B and C were produced by TestGris according to Danish feeding standards.

[0100] 4111 Danbred crossbred piglets were used, with a weaning age of 25±3 days and an average weight of 6.4 kg. 64 double pens were used. Approximately 2*32 piglets were used in each double pen. The double pens were assigned to one of four diets.

[0101] Output measurements were average daily gain (ADG), feed intake (FI), and feed utilization (FU) (also known as feed conversion ratio (FCR)). The results are shown in Table 1.

[0102] [Table 5]

[0103] Overall, the piglets remained in good health throughout the study.

[0104] It was concluded that ADG tended to be significantly affected by dietary treatment in Phase A, with the highest ADG in Diet 4. FI was not affected by dietary treatment, but FU was significantly higher in Diet 4 compared to Diet 1. The different diets fed in Phase A did not affect performance in Phases B or C.

[0105] Example 7 Parameters such as water-holding capacity (WHC) and viscosity are important in fiber products. A relatively high water-holding capacity in a fiber product helps ensure satiety in the animal and optimal stool consistency. Viscosity is the fiber property that often causes concern, especially in poultry feeding, where high viscosity is undesirable as it can slow down nutrient absorption and give pathogens more room to grow in the intestine. Water-holding capacity is measured as the amount of water that a biomass can hold against gravity without releasing free water, and is expressed in kg water / kg biomass.

[0106] Water holding capacity (WHC) The WHC was measured in various dry fiber products including raw soybean hulls and the finished fiber component according to the invention, i.e. pectinase-treated soybean hulls and barley, wheat, corn, and soybean meal (SBM). All samples were first disintegrated through a 500 μm sieve. The results are shown in FIG. 8 and show that the product of the invention has a satisfactorily high WHC, as also characterizes many other fiber products.

[0107] viscosity For viscosity measurements, the following method was applied: all samples were milled through a 500 μm sieve. 1 g of sample and 20 mL of 0.05 M phosphate buffer (pH 7) were incubated at 40° C. for 0.5 h. The samples were then centrifuged at 500 rpm (31 G) for 10 min in an Allegar X-22R centrifuge, Beckman Coulter, prior to rheological analysis. The samples were analyzed with a Discovery HR-3 rheometer (TA Instruments). The finished fiber component according to the invention, i.e. pectinase treated soybean hulls, is compared with raw soybean hulls, sugar beet pulp, wheat bran and Easy Fibre (a blend of rape straw and wheat straw). The results are shown in FIG. 9 and show that the product of the invention has a reduced viscosity compared to raw soybean hulls, wheat bran, sugar beet pulp and Easy Fibre. The relevant shear rates in the intestine are approximately 1 to 10 s -1 It is between. [Table 6] [Table 7] [Table 8]

Claims

1. A feed or food ingredient derived from soybean hulls, said feed or food ingredient comprising dietary fibre derived from soybean hulls in the form of soluble and insoluble polysaccharides, a portion of the dietary fibre derived from the soybean hulls being degraded by one or more carbohydrases selected from mannanase, pectinase, xylanase, glucanase and cellulase to provide the feed or food ingredient comprising 4% by mass or more of said oligosaccharides DP3-30, calculated assuming no soybean oligosaccharides raffinose, stachyose and verbascose, said feed or food ingredient further comprising yeast, with the proviso that said feed or food ingredient does not comprise more than 5% by mass of monosaccharides.

2. 2. The feed or food ingredient according to claim 1, wherein part of the dietary fibre derived from soybean hulls is degraded by the one or more carbohydrases into oligosaccharides (oligosaccharides DP3-30), providing a prebiotic effect.

3. A feed or food ingredient as described in claim 1 or 2, comprising 5% or more by weight of oligosaccharides having 3 to 30 monomer sugar units (oligosaccharides DP3-30), for example 6% or more by weight, 7% or more by weight, 8% or more by weight, 9% or more by weight, 10% or more by weight, 12% or more by weight, 15% or more by weight, or 20% or more by weight of oligosaccharides having 3 to 30 monomer sugar units (oligosaccharides DP3-30).

4. A feed or food ingredient according to any one of claims 1 to 3, comprising a yeast selected from among a Saccharomyces cerevisiae strain, such as spent brewer's yeast and spent distiller's yeast and spent yeast from wine production, bioethanol yeast or spent yeast from bioethanol production, baker's yeast and yeast strains which ferment C5 sugars.

5. A feed or food ingredient as described in any one of claims 1 to 4 comprising yeast in an amount of 0.05 to 10%, for example 0.10%, 0.15%, 0.20%, 0.25%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5, 5%, 6%, 7%, 8% or 9%.

6. A feed or food ingredient described in any one of claims 1 to 5, further comprising the oligosaccharides raffinose, stachyose and verbascose present in the soybean husk being completely or partially degraded.

7. 7. The feed or food ingredient according to any one of the preceding claims, further comprising one or more microorganisms and / or metabolic products of carbohydrate degradation of the soybean hulls by said one or more microorganisms.

8. 8. The feed or food ingredient according to claim 7, further comprising lactic acid bacteria and / or metabolic products of carbohydrate degradation of the soybean hulls by lactic acid bacteria.

9. A method for producing a feed or food ingredient according to any one of claims 1 to 8, comprising the following steps: - mixing the soybean hulls with one or more carbohydrases selected from mannanase, pectinase, xylanase, glucanase, and cellulase; - adding yeast to the mixture of soybean hulls and carbohydrase; - hydrolyzing said mixture at a temperature of from 20 to 60°C for a period of from 1 to 48 hours under conditions in which the DM content is less than or equal to 55% by weight; - optionally completely inactivating the activity of said one or more carbohydrases, - isolating said feed or food ingredient A method comprising:

10. The method of claim 9, wherein the soybean hulls are disintegrated prior to mixing with the one or more carbohydrases.

11. The method according to claim 9 or 10, comprising adding a yeast selected from among Saccharomyces cerevisiae strains, such as spent brewer's yeast and spent distiller's yeast and spent yeast from wine production, bioethanol yeast or spent yeast from bioethanol production, baker's yeast and yeast strains fermenting C5 sugars, to the mixture of soybean hulls and carbohydrase prior to the hydrolysis step.

12. The method of claim 9, further comprising adding one or more microorganisms to the mixture of soybean hulls and carbohydrase prior to the hydrolyzing step.

13. The method of claim 12, further comprising adding lactic acid bacteria to the mixture of soybean hulls and carbohydrase prior to the hydrolyzing step.

14. The method of any one of claims 9 to 13, further comprising adding alpha-galactosidase to the mixture of soybean hulls and carbohydrase prior to the hydrolyzing step.

15. 9. A feed or food product or a dietary supplement comprising from 0.5 to 99% by weight of a feed or food ingredient according to any one of claims 1 to 8.

16. 16. A feed product or dietary supplement as claimed in claim 15 for use in a diet for production animals, such as a diet for improving performance.

17. 17. A feed product according to claim 15 or 16 for use in diets for production animals requiring prebiotic oligosaccharides, preferably newborn and young animals, such as piglets, calves and chickens.

18. 9. Use of a feed ingredient according to any one of claims 1 to 8 in diets for production animals, preferably newborn and young animals, such as piglets, calves and chickens.

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