Process for preparing a beverage or beverage component from brewer's spent grains

Abstract

A process prepares a beverage or beverage component. The process includes an enzymatic treatment of brewer's spent grain including addition of one or a combination of enzymes with alpha-amylase, gluco-amylase, cellulase, xylanase, protease and/or beta-glucanase activity and fermentation by a strain of lactic acid bacteria. The combination of enzymes and enzymatic treatment conditions is such that the lactic acid bacteria produce 4.5 g/L lactic acid and metabolise sugar such that the resulting fermented broth contains less than 2.5% w/w and preferably less than 0.5% w/w residual sugar or the lactic acid bacteria produce 4.5 g/L lactic acid and metabolise sugar such that the resulting fermented broth contains at least 2.5% w/w residual sugar.

Claims

1. A process for preparing a beverage or beverage component comprising: hydrolyzing brewer's spent grain by the addition of enzymes with alpha-amylase, gluco-amylase, cellulase, xylanase, protease and beta-glucanase activity, resulting in saccharified brewer's spent grain, fermenting the saccharified brewer's spent grain by a strain of lactic acid bacteria to obtain a fermented broth, wherein the steps of hydrolyzing and fermenting are combined in one step performed for between 15 and 24 hours at a temperature between 25 and 37° C. such that: said lactic acid bacteria produce 4.5 g/L lactic acid and metabolise sugar such that the resulting fermented broth contains less than 2.5% w/w residual sugar; filtering the fermented broth and collecting the permeate to obtain the filtered beverage or filtered beverage component, or homogenizing the fermented broth to obtain the homogenized beverage or homogenized beverage component, and supplementing the filtered beverage or filtered beverage component, or the homogenized beverage or homogenized beverage component, with a probiotic microorganism being a lactic acid bacteria, wherein the beverage or beverage component comprises at least 1.4% (w/v) of water-extractable arabinoxylans (WEAX).

2. The process according to claim 1, the residual sugar exclusively originating from the brewers' spent grain.

3. The process according to claim 1, wherein brewer's spent grain is treated with enzymes to solubilize arabinoxylans.

4. The process according to claim 1, comprising the step of mixing the beverage component with a diluent, compound or another beverage to obtain a beverage.

5. The process according to claim 1, wherein the final beverage is supplemented by a probiotic microorganism after pasteurization, the probiotic microorganism consisting of Lactobacillus rhamnosus.

6. A beverage or beverage component obtained by fermentation of saccharified brewer's spent grain and homogenization of fermented broth and spent grain, the beverage or beverage component comprising at least 1.4% (w/v) of water-extractable arabinoxylans (WEAX), proteins in a level sufficiently high such that at least 12% of the total caloric value of the beverage or beverage component originates from proteins therein, the beverage or beverage component supplemented with a probiotic microorganism of lactic acid bacteria.

7. The beverage or beverage component according to claim 6, being a low calorie/energy beverage having a caloric value of: less than 20 kcal/100 mL, or at least 20 kcal/100 g.

8. The beverage or beverage component according to claim 7, having a sugar content of less than 0.5% w/v, or at least 0.5% w/v and less than 2.5% w/v or at least 2.5% w/v.

9. The beverage or beverage component according to claim 6, having a sugar content of less than 0.5% w/v, or at least 0.5% w/v and less than 2.5% w/v, or at least 2.5% w/v.

10. The beverage or beverage component according to claim 6 comprising, 3% (w/v) of water-extractable arabinoxylans (WEAX).

11. The beverage or beverage component according to claim 6, having a fat content of less than 1.5 w %.

12. The beverage component according to claim 6 wherein the beverage or beverage component is lactose free.

13. The beverage or beverage component according to claim 6 having a fiber content of at least 1.5 g per 100 kcal of beverage or beverage component.

14. The beverage or beverage component according to claim 13, being a low calorie/energy beverage having a caloric value of: less than 20 kcal/100 mL, or at least 20 kcal/100 g.

15. The beverage or beverage component according to claim 13, having a sugar content of less than 0.5% w/v, or at least 0.5% w/v and less than 2.5% w/v, or at least 2.5% w/v.

16. A beverage or beverage component obtained by fermentation of saccharified brewer's spent grain and filtering the fermented broth from the spent grain and supplemented with a probiotic microorganism of lactic acid bacteria, wherein the beverage or beverage component comprises at least 1.4% (w/v) of water-extractable arabinoxylans (WEAX).

17. The beverage or beverage component according to claim 16, being a low calorie/energy beverage having a caloric value of: less than 20 kcal/100 mL, or at least 20 kcal/100 g.

18. The beverage or beverage component according to claim 16, having a sugar content of less than 0.5% w/v, or at least 0.5% w/v and less than 2.5% w/v, or at least 2.5% w/v.

Description

BRIEF DESCRIPTION OF THE FIGURES

(1) FIG. 1 shows an example of simultaneous saccharification and fermentation (SSF) process. Bacterial fermentation causes an increase in lactic acid and titratable acidity in the media. There is an initial increase in sugar concentration due to the saccharification process. After a short lag phase, bacteria begin consuming the sugar, and its concentration decreases. Fermentation is stopped when .sup.˜4.5 g/L lactic acid are produced (black arrow). In this example, the enzyme mix and initial sugar in BSG are such that no residual sugar is left at the point of stopping the fermentation.

DEFINITIONS

(2) Barley is the main raw material used for the production of beer. However, other cereals such as corn or rice are typically used together with malted barley. During the brewing process the starchy endosperm of these cereals is subjected to enzymatic degradation, resulting in the liberation of fermentable (maltose and maltotriose, and a minor percentage of glucose) and non-fermentable carbohydrates (dextrins), proteins, polypeptides and amino acids. The thus produced medium (which will be fermented into beer by the action of yeast) is known as wort. The insoluble grain components (comprising mainly the grain coverings) is the brewers' spent grain (BSG). In traditional brewing employing a lauter tun, the BSG components play an important role as they form the bed through which the mash is filtered to produce wort. Therefore, the initial milling of the malt must be such that the grain coverings remain intact so as to form an adequate filter. Today, while many small or craft breweries still use this method of mash filtration, many larger breweries employ a mash filter which relies less on the filtration function of the BSG and thus malt can be milled more extensively.

(3) The brewer's spent grain contains all the solids that have been separated from the wort by filtration; it includes what is left of the barley malt and the adjuncts. The spent grain consists mainly of the pericarp and hull portions of the barley and of non-starchy parts of corn, provided corn grits were used as an adjunct. Brewer's spent grain is a lignocellulosic material typically comprising lipids, lignin, proteins, cellulose, hemicellulose and some ash. For the description and claims of this invention the wording “brewer's spent grain” (BSG) will be used in accordance with the definition here above.

(4) Extract in the brewing context, and in the context of this invention, refers to soluble solids extracted into a liquid phase during mashing (for brewing) or SSF (this invention). It is understood that the overwhelming majority of these solids are fermentable sugars, like maltose (in brewing) or glucose (in brewing and SSF of this invention).

(5) Product water refers to water used in the brewing process, that has suffered a defined and standard process for making it suitable for consumption.

(6) Nutritional definitions as defined by the European Commission (http://ec.europa.eu/food/safety/labelling_nutrition/claims/nutrition_claims/index_en. htm), see Table below:

(7) TABLE-US-00001 Nutritional claim Definition Low energy <20 kCal per 100 g Fat free <0.5% fat content Low fat <1.5% fat content Very low salt <0.4% salt content Source of fiber >3% fiber content OR > 1.5 g fiber per 100 kCal Sugars-free <0.5% w/v sugar content Low sugars <2.5% w/v sugar content ‘With no added Does not contain any added mono- or disaccharides sugars’ or any other food used for its sweetening properties. High in fiber >6% fiber content OR > 3 g fiber per 100 kCal Source of protein >12% of the energy provided by protein High in protein >20% of the energy provided by protein

(8) Digestion of AX either enzymatically or otherwise results in an increase of the soluble fraction of arabinoxylans (WEAX). This fraction is responsible for most of the health-promoting effects of arabinoxylans. Among the many positive effects WEAX have on health we find: 1. reduction of postprandial glucose levels in individuals with compromised glucose metabolism (Lu et al. 2004; Garcia et al. 2007) 2. tumor suppressing activity (Cao et al. 2011) 3. reduction of obesity, cholesterol levels and restoration of beneficial gut bacteria in high fat diets (Neyrinck et al. 2011) 4. Immune-enhancing effects (Zhou et al. 2010) 5. prebiotic effects, including promoting healthy gut bacteria and short chain fatty acid in distal colon (Cloetens et al. 2010; Sanchez et al. 2009)

(9) Additionally, there is evidence that preparations of arabinoxylans from brewer's spent grains (BSG-AX) can exert the same prebiotic effects as the better-studied wheat-derived arabinoxylans, namely: 6. BSG-AX are not absorbed in the small intestine and reach the colon (Teixeira et al. 2017); BSG-AX promote proliferation of gut bacteria, particularly beneficial species like, for example, those of the Bifidobacteria genus, and BSG-AX promote the production of short chain fatty acids by said bacteria (Reis et al. 2014)

(10) The documented effects listed above were elicited by the following dosages: (1) 0.12 g/kg body weight/day, (2) 0.4 g/kg body weight/day, (3) 10% of diet, (4) 0.1 g/kg day, (5) 0.14 g/kg weight/day and 0.6% (w/v), (6) 0.6 g/kg body weight/day

(11) Additionally, a patent concerning the use of soluble arabinoxylans extracted from wheat (Ekhart et al. 2012), recommends that a daily dosage of 0.08 g/kg day would be adequate to obtained the claimed health effects, namely prebiotic effect and decrease of symptoms associated with high-fat diets.

(12) European Food Safety Authority has concluded that there is a cause effect relationship between the consumption of wheat arabinoxylan and the reduction of postprandial glucose levels (Efsa Panel on Dietetic Products 2011). Based on the provided evidence EFSA suggests that to obtain the claimed effect, 4.8% w/w of consumed carbohydrate should be soluble arabinoxylans. For a healthy 70 kg adult with an average 2200 kcal daily intake (EFSA Panel on Dietetic Products Nutrition and Allergies 2013), of which 45% are carbohydrates (EFSA Panel on Dietetic Products Nutrition and Allergies 2010), this corresponds to 0.17 g/kg body weight/day.

(13) It is therefore considered that no less than 0.1 g/kg body weight/day, is a sufficient dose of WEAX to have positive health effects.

(14) The fibre-solubilization and saccharification enzyme process described here results in a beverage, beverage ingredient or food ingredient with no less than 1.4% (w/v) soluble arabinoxylans.

(15) Finally, lactose free refers to a product that contains no trace of this compound. The present invention refers to a beverage produced through the fermentation of BSGs, therefore containing no dairy product and thus lactose free.

DETAILED DESCRIPTION OF A PREFERRED EMBODIMENT

(16) The process according to the present invention generally comprises the steps of: Providing brewer's spent grain; Performing saccharification and fibre solubilization by enzymatic treatment of the brewer's spent grain; Fermenting the saccharified brewer's spent grain with lactic acid bacteria and/or acetic acid bacteria and/or probiotics to obtain a fermented broth; and filtering the fermented broth and collecting the permeate to obtain the beverage or beverage component; or homogenizing the fermented broth to obtain the beverage or beverage component.

(17) The brewer's spent grain is preferably obtained from a regular beer production process, wherein malt and potentially some adjuncts such as corn, rice, sorghum, wheat, barley, rye, oat or combinations thereof are mixed with water to form a mash wherein enzymes—either originating from the barley malt or added separately to the mash—are allowed to break down starch into fermentable sugars, typically a mixture of glucose, maltose and maltotriose. At the end of the mashing, the mash is filtered to obtain a fermentable wort that is further processed in to beer. The retentate of the mash filtering is the brewer's spent grain (BSG).

(18) BSG comprises the seed coat-pericarp-husk layers that covered the original barley grain. BSG's composition mainly comprises fibers, which are non-starch polysaccharides (NSP; hemicellulose in the form of arabinoxylans (AX) and cellulose) and significant quantities of proteins and lignin, with arabinoxylans (AX) typically constituting the most abundant component. Therefore, BSG is basically a lignocellulosic material. Fiber constitutes about half of the BSG composition on a dry weight basis, while proteins can constitute up to 30% of the dry weight basis. This high fiber and protein content makes BSG an interesting raw material for food applications.

(19) As would be expected, cellulose (β-(1,4)-linked glucose residues) is another abundant polysaccharide in BSG. Certain levels of (1-3,1-4)-β-D-glucan may also be present. The most abundant monosaccharides in BSG are xylose, glucose, and arabinose, while traces of traces of rhamnose and galactose have also been found.

(20) The protein content of BSG typically is present at levels of approximately 30% per dry weight basis. The most abundant are hordeins, glutelins, globulins and albumins. Essential amino acids represent approximately 30% of the total protein content, with lysine being the most abundant, while non-essential amino acids in BSG constitute up to 70% of the total protein content. This is significant because lysine is often deficient in cereal foods. In addition, BSG also contains a variety of minerals elements, among which silicon, phosphorus, calcium and magnesium are the most abundant.

(21) The BSG obtained from a lager beer production process typically comprises hemicellulose (20-25 w % on dry matter); cellulose (12-25 w % on dry matter); protein (19-30 w % on dry matter); lignin (12-28 w % on dry matter); lipid (ca. 10 w % on dry matter); ash (2-5 w % on dry matter); and low amounts of fructose, lactose, glucose and maltose.

(22) The BSG is highly nutritious and very sensitive for spoilage by micro-organisms, hence heat treating of the BSG is desired to increase the shelf life. In this sense, the high water content of BSGs in the moment of their production (wort filtration), which is in the range of 75% (25% total solids), increases the instability of the material. For this reasons preferably fresh spent grains are used in the process of the present invention, and/or BSGs are stabilized or treated for sterilization, preferably by boiling.

(23) In a process according to the present invention, BSGs, preferably as produced during the brewing process (in the range of 25% total solid content), and more preferably collected just after their production, are mixed with distilled water, or preferably hot product water, to a final dry matter content of between 6 and 10%, more preferably between 8 and 9%, and subsequently treated for stabilization, for example by heat treatment such as by boiling for 60 minutes. Subsequently, the mixture of BSGs and water is exposed to fibre solubilization, saccharification and fermentation, preferably to a simultaneous process of saccharification and fermentation (SSF). Commercial enzymatic products used for the fibre solubilization and saccharification of the BSG in the present invention will have at least one of following activities: xylanase (including endo-xylanase); cellulase; glucanase (including beta-glucanase); glucoamylase, protease, and or admixtures thereof. Preferably, the enzymatic mixture use will contain starch, dextrin, protein and fiber degrading activities. More preferably, these activities will comprise gluco-amylase, pullulanase, alpha-amylase, beta-glucanase, xylanase and protease. Enzyme treatment with xylanase and protease solubilizes WUAX and increases the levels of health-promoting WEAX.

(24) The choice of enzymes and conditions will affect the amount of sugar released from fiber in the saccharification process. Because bacterial fermentation is stopped after production of a determined amount of acid, the amount of sugar released will affect the amount of residual sugar left after fermentation. Example 1 shows a combination of enzymes that results in a relatively low release of sugars from fiber.

(25) As examples of such enzyme treatment, experiments were done by adding to a mixture of BSGs and water the following commercial products:

(26) TABLE-US-00002 Enzyme combination 1 Declared enzymatic Commercial Product Supplier activities Dose Ultraflo FABI Novozymes Beta-glucanase 100 ppm Endo-xylanase Alpha-amylase Attenuzyme PRO Novozymes Gluco-amylase 500 ppm Pullulanase Alpha-amylase Acellerase ® 1500 DuPont Exoglucanase 1500 ppm  Endoglucanase Hemi-cellulase Beta-glucosidase Alcalase ® 2.4 L Novozymes Protease (serine 200 ppm endopeptidase)

(27) TABLE-US-00003 Enzyme combination 2 Declared enzymatic Commercial Product Supplier activities Dose Ultraflo FABI Novozymes Beta-glucanase 100 ppm Endo-xylanase Alpha-amylase Attenuzyme PRO Novozymes Gluco-amylase 500 ppm Pullanase Alpha-amylase Allzyme proteases 12500 ppm  Amylase Xylanase Beta-glucanse Pectinase cellulase Phytase Alcalase ® 2.4 L Novozymes Protease (serine 200 ppm endopeptidase)

(28) TABLE-US-00004 Enzyme combination 3 Declared enzymatic Commercial Product Supplier activities Dose Allzyme proteases 10000 ppm Amylase Xylanase Beta-glucanse Pectinase cellulase Phytase Alcalase ® 2.4 L Novozymes Protease (serine  200 ppm endopeptidase)

(29) Table 1 shows how the combination of enzymes and saccharification time can be used to tailor the amount of sugar released from fiber. Incubation temperature was 55′C and pH was 5.5 in all reactions.

(30) TABLE-US-00005 TABLE 1 Effect of diffetent enzyme combinations and incubation time on saccharification Extract Enxyme Incubation time Initial extract Final extract released combination (hrs.) (g/100 mL) (g/100 mL) (g/100 mL) 1 72 1.8 3.7 1.9 2 72 1.8 4.5 2.7 3 96 (Allzyme), 1.8 5.5 3.7 24 (Alcalase)

(31) After hydrolysis, a fermentable broth is obtained that is subsequently fermented with lactic acid bacteria and/or acetic acid bacteria and/or probiotics. Preferably, such microorganisms are added during the hydrolysis, thus performing a simultaneous saccharification and fermentation process (SSF).

(32) Examples of lactic acid bacteria include:

(33) TABLE-US-00006 Species Strain Metabolism Origin L. amylovorus AB32 Homofermentative Sourdough L. amylovorus AB36 Homofermentative Sourdough L. brevis WLP672 Heterofermentative L. brevis JJ2P Heterofermentative Porcine L. paracasei CRL431 Heterofermentative Infant faeces L. casei R10 Heterofermentative Cheese L. casei H2 Heterofermentative Human L. crispaticus AB19 Homofermentative Sourdoug L. delbreuckii WLP677 Homofermentative L. fermentum AB15 Heterofermentative Sourdough L. fermentum AB31 Heterofermentative Sourdough L. fermentum F23 Heterofermentative Sourdough L. gallinarum AB13 Homofermentative Sourdough L. plantarum F6 Heterofermentative Sourdough L. plantarum F10 Heterofermentative Brewery L. plantarum F21 Heterofermentative Sourdough L. plantarum R11 Heterofermentative Cheese L. plantarum R13 Heterofermentative Cheese L. reuteri AB38 Heterofermentative Sourdough L. reuteri DSM20016 Heterofermentative Human intestine L. reuteri Ff2 Heterofermentative Porcine L. reuteri hh1P Heterofermentative Porcine L. reuteri R12 Heterofermentative Cheese L. rhamnosus C7 Homofermentative Cheese L. rhamnosus C8 Homofermentative Cheese L. rhamnosus C9 Homofermentative Cheese L. rhamnosus GG Homofermentative Human gut L. sakei AB3a Heterofermentative Sourdough L. vaginalis AB11 Heterofermentative Sourdough Leuconostoc TR116 Heterofermentative Sourdough citreum L. holzapfelii AB4 Heterofermentative Sourdough Leuconostoc E11 Heterofermentative Sourdough lactis Leuc. DSM20240 Heterofermentative Root beer Mesenteroides Weissella cibaria MG1 Heterofermentative Sourdough

(34) Examples of acetic acid bacteria include G. oxydans and K. xylinus.

(35) Preferably, the strains L. planetarum F10 and L. rhamnosus LGG are preferred as selected to provide desirable organoleptic properties. Possibly, a probiotic strain is added at the end of the process of production of the beverage defined in the present invention.

(36) Hydrolysis of the BSG is performed for at least 12 hours, preferably 24 hours at a temperature in function of the enzyme(s) used (typically about 55′C), to ensure solubilization of arabinoxylans and increase in the level of WEAX to health-promoting levels of at least 1.4% (w/v). Hydrolysis is followed by a 8 to 24 hours of fermentation at about 25 to 37′C, preferably at 30° C. Preferably, the hydrolysis and fermentation steps are combined in one step (SSF) and performed during between 15 and 24 h at a temperature between 25 and 37° C., more preferably during 20 h at a temperature of 30° C. Aerobic and static conditions are used during the fermentation or SSF process.

(37) The fermentation or SSF is followed by critical parameters such us pH, extract, total acidity (TTA) and concentration of reducing sugars. The process is considered to be finished when, for example, 4.5 g/L lactic acid are produced by the bacteria, or the total titrateble acidity (TTA) of the broth is such that 10 mL of it are titrated to pH 7 by 3 mL a 0.1M solution of sodium hydroxyde, and, more preferably, a drop of between 0.2 and 0.4 pH units from the initial pH is observed. Alcohol concentration in the fermented broth is also measured. Aerobic and static conditions are used to ensure a low alcohol concentration, below 0.20%, preferably below 0.15%, and more preferable below 0.10% in the fermented broth.

(38) FIG. 1 shows an example of an SSF. Bacterial fermentation causes an increase in lactic acid and titratable acidity in the media. There is an initial increase in sugar concentration due to the saccharification process. After a short lag phase, bacteria begin consuming the sugar, and its concentration decreases. Fermentation is stopped when .sup.˜4.5 g/l lactic acid are produced (black arrow). In this example, the enzyme mix and initial sugar in BSG are such that no residual sugar is left at the point of stopping the fermentation.

(39) The lactic acid fermentation or SSF process is arrested by cooling the ferment to a temperature lower than 18° C., or, preferably, heating the ferment to a temperature above 50° C.

(40) The above described fermented broth can follow two different subsequent processes, leading to two different types of beverages or beverage components: 1. Fermented broth can be filtered to produce a filtered beverage by the following process: The fermented base is swirled to re-suspend settled particles. Solid (insoluble) particles are allowed to settle, preferably by centrifugation. The resulting supernatant is filtered, preferably through mash filters. Further filtration steps are possible to reduce the size of particles in the final beverage. According to the extent of saccharification and the consequent level of residual sugar of the beverage, this beverage can have either of the following nutritional claims (see definitions): Low energy, fat-free, sugars-free, very low salt content; or low energy, fat-free, low in sugar, very low salt content; or fat-free, very low salt content and ‘With no added sugars’. 2. Fermented broth can be homogenized to produce a beverage by the following process: The fermented base is swirled to re-suspend settled particles. The mixture is then blended, preferably by an industrial blender, until a homogenous mixture is obtained. According to the extent of saccharification and the consequent level of residual sugar of the beverage, this one can have either of the following nutritional claims (see definitions): High in fibre, fat-free, sugar-free, high in protein, very low salt content; or High in fibre, fat-free, low in sugar, high in protein, very low salt content; or High in fibre, fat free, high in protein, very low salt content and ‘No added sugars’.

(41) By filtering the fermented broth, a beverage, beverage component or food component (type 1) can be obtained that is low in energy (<20 kcal/100 mL) and/or fat free (<0.5%) and/or sugar free (<0.5%) or low in sugar (2.5% w/v) and/or very low in salt content (<0.4%) and/or contains sufficient levels of health-promoting soluble arabinoxylans (no less than 1.4% w/v, preferably 3%). A 500 mL serving of said beverage would provide 70 g of soluble arabinoxylans, or 0.1 g/kg body weight for a 70 kg adult person.

(42) By homogenizing a beverage or beverage component (type 2) the fermented broth, a beverage, beverage component or food component (type 2) can be obtained that is low in fat content (<1.5%) and/or sugar free (<0.5%) or low in sugar (2.5% w/v) and/or high in fiber content (>1.5 g fiber/100 kcal, preferably >3 g fiber/100 kcal) and/or sufficient levels of health-promoting soluble arabinoxylans (no less than 1.4% w/v, preferably 3%) and/or high in protein (>12%, preferably >20% of the energy provided by proteins) and/or very low in salt content (<0.4%). A 500 mL serving of said beverage would provide 70 g of soluble arabinoxylans, or 0.1 g/kg body weight for a 70 kg adult person.

(43) Since no dairy product is used in the described process, the beverage or beverage component obtained by a process according to the present invention is consequently lactose free.

(44) The beverage can be consumed as such or can be used as a beverage component and mixed with one or more other components prior to consumption. Such components can be beverages as for example a fruit juice. The beverage can be used as a food component or food additive for foodstuffs such as: pasta products, breads and sourdoughs, cereals and cereal products, baked goods and cookies.

(45) The final beverage, beverage component or food component obtained by the process described in this invention can be exposed to stabilization treatments, preferably pasteurization, preferably at 70 C during 30 min. Additionally, the final beverage or beverage component can be supplemented by the addition of probiotic microorganisms, preferably lactic acid bacteria.

REFERENCES

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