DE2320425A1 - PROCESS FOR OBTAINING A LIQUID OR DRIED DEGRADATION PRODUCT FROM CEREALS - Google Patents

Description

Oasis 3/103
Dr.Ve/Ba

CH. Boehringer Sohn, Ingellieim / Rhine 1

Process for obtaining a liquid or dried

Digestion product made from cereals

The invention relates to a method using of microbial and / or vegetable enzymes as well as a low malt bulk fraction one technologically and economically favorable digestion of raw cereal fruit possible. Depending on the area of application, the digestion product can be in liquid or dried (e.g. spray-dried) Form of its further processing. It can be used in particular as wort, wort syrup or wort - find powder for brewing purposes. Furthermore, this can Digestion product as a raw material for various alcoholic and non-alcoholic beverages, for malt sweets and other sweets, dietetic and children's food, microbiological Nutrient media, can be used as a baking additive, among others. The invention also relates to a method for carrying out this method suitable special enzyme mixture.

40984 5/0933

The classic mashing processes for the production of beer wort are based on the raw material barley lard, which in one upstream, relatively lengthy and complex malting process is made from brewing barley. The partial replacement of the malt with cheaper, unmalted barley or other Unmalted cereals (raw fruit) are possible and have been used in many countries for a long time. Due to still "existing legal Regulations, this economically advantageous use of raw fruit in German breweries is currently still in exceptional cases, E.g. the production of export beers is limited. As the technical and patent literature shows, raw fruit spices are found however, there is growing interest worldwide, especially as their use in products other than beer is also being considered.

The replacement of the malt with raw fruit during mashing is only possible to a very limited extent without the addition of enzymes (cf. WIEG, A.J .: Die Starke 22, 200-210 (1970). In order to be able to increase the raw fruit bulk portion, since some years working methods suggested the addition of enzymes - in particular ot-amylase, amyloglucosidase (= Glucoamylase) and proteases - to be provided. The vast majority of these processes keep mash raster at rising temperatures a, as it is known from the traditional infusion and decoction processes of the brewery (see DOS 1 422 288; DOS 1,642,748; U.S. Patent Nos. 2,790,718 and 3,081,172; KLOPFER, W.J .: The Brewery Technician 20, 1-4 (1968); MACEY, A. i.a .: EBC-Oongress Proc. 1967, pp. 283-294). Other Methods that have become known consist of complicated alternating heating and cooling phases as well as more fractional ones

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Enzyme addition (see DOS 1 903 400) or they contain upstream higher temperature levels for non-enzymatic digestion at boiling temperatures, under pressure (see DOS 2 124 846; DOS 1 911 571) or with the action of acid (see Brit. Patent 1 203 623). Such working methods have the specialist part high energy expenditure, difficult controllability of the thermal digestion process and the Formation of undesirable flavors and colors. When using acids for hydrolysis it also still makes itself their corrosive effect on the exclusion devices is unpleasant noticeable.

In the enzymatic digestion of raw fruit, low molecular weight is formed (Fermentable) sugars from the high molecular starch mainly by saccharifying enzymes, such as Amyloglucosidase and ß-amylase, the glucose and maltose units from the non-reducing chain end of the glucose polymerization split off. The prerequisite for an optimal effect of these enzymes is the presence of a sufficient number of non-reducing ones Chain ends; In the case of the high molecular starch components, which are initially undestroyed, this is amylose and amylopectin does not. First the oc-amylase is chopped up the amylose and amylopectin molecules coarsely into dextrins and creates many new non-reducing ends, that is, points of attack for further saccharification.

It has now been found that a high quality digestion product can be obtained by means of a novel mashing process can be obtained, which by using a new enzyme mixture with minimal addition of malt a very cost-saving Processing of raw fruit (e.g. barley, rye, wheat, maize, rice, oats or millet) allowed.

/ 4. 409845/0933

The method according to the invention works using a specially developed enzyme mixture with only two mash racks, the first of which is at a higher temperature than the second. The first mash rest is around 67-73 ° C and is used for liquefaction and dextrination with a small amount of malt added. The second Maisch radical has the task of reducing existing proteins and to saccharification, the dextrins formed during the first mash rest, it is carried out at about 56 ⁰ C with the addition of Enzymgeinisches invention.

The enzyme mixture to be added according to the invention only at the second mash rest was developed with the purpose of keeping the number of pH and temperature changes as small as possible with the best possible digestion effect and thus simplifying the technology of the mashing process. In view of this objective, an enzyme mixture was put together, the enzymes of which match as much as possible in terms of their pH and temperature optimum. The optimum temperature of the mixture according to the invention is between 34 ⁰ C and 62 ⁰ C, preferably at about 5UE ° C and the optimum pH of from 4.5 to 5> 5j preferably at pH 5> 0th These are therefore also the process conditions for the second mash rest.

It was also recognized that, contrary to the prevailing opinion for an enzyme mixture which is to be used for the preparation of wort from oil to 96% raw fruit, an addition of α-amylase is superfluous if the above process conditions are observed. In this case

409845/0933

A minimal addition of malt is sufficient to achieve optimal dextrination and normal iodine levels before the first mash rest is over. With the <x -amylase alone, which comes from the low malt content, so many dextrin pieces with non-reducing chain ends are evidently formed that the saccharifying amyloglucosidase is already in a state of substrate saturation and adding ct-amylase would not further increase the sugar formation rate. This result is surprising, because in the literature p-err.ee the α-amylase is seen as a limiting factor in raw fruit processing (cf. RA Latimer et al, Inst. Brew. Proc. Conf. - ±, pp. 111-126 ( 1966); WJ Klopper, Der Brauereitechniker 20, pp. 1 - 4. (1968).

basically An enzyme mixture according to the invention contains no ^xy demgemätf

α-amylase. On the other hand, it contains the necessary components Amyloglucosidase and at least one microbial and / or vegetable protease. The presence also has a beneficial effect of cellulases or hemicellulases and pectinases. Such enzymes are usually as secondary activities in the Mixture contain so many that the amylolytic and proteolytic enzymes are effectively supported and good filterability the mash is guaranteed even with flour-fine grinding. Only in exceptional cases in which the secondary activities not sufficient (for example when processing rye of certain origins) is a small addition of pentosanase and / or β-glucanase is expedient.

3s has also been found to have an optimal effect of such an enzyme mixture can be achieved if the mixture has an activity ratio of the amyloglucosidase units to protease units of 1: 0.03 to 0.5.

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For example, the enzyme mixture can contain about 500 to 1500 units per gram of amyloglucosidase, as well as 50 to 250 units contained per gram of proteases.

A typical standardized enzyme mixture according to the invention has, for example, the following main activities:

1,000 amyloglucosidase units per g 100 protease units per g

and is standardized to these activity values with an enzyme-free substance.

Of course, a more dilute or more concentrated enzyme mixture also has the same effect, if only that The above-mentioned ratio of activities is observed and the amount used is adjusted accordingly. Making a Enzyme mixture according to the invention can be carried out in such a way that amyloglucosidase with at least one protease mixes microbial or vegetable origin, if desired also adds cellulases, hemicellulases and / or pectinases and by mixing with an enzyme-free carrier substance, for example starch or lactose, standardized.

The specified enzyme activities have been defined and determined as follows:

Amyloglucosidase activity

An amyloglucosidase unit corresponds to the amount of enzyme that exposes at pH 4.4 and 25 ⁰ C in one minute from soluble starch as vieIe ^ groups that they meet at a iodometric determination ^ MoI glucose.

^ reducing .- ".. ■ / 7

409845/0333

The determination is carried out as described by H.J. Pieper: microbial amylases in alcohol production, Pp. 48-49, Ulmer Verlag, Stuttgart, 1970.

Protease activity

A protease unit is the amount of enzyme which 5.0 and 3O ⁰ C so many trichloressigsäurelösliche cleavage products formed at a pH in a minute of denatured hemoglobin that these measured by the color reaction according to Folin-Ciocalteu correspond to a -mol tyrosine.

Except for the above defined conditions (pH 5.0 and 30 ^° C.), the determination is carried out according to R. Ruyssen, - (Ed.): Symposium on Pharmaceutical Enzymes and their Assay, pp. 120-127, Univers. Press, Ghent, 1969.

α-amylase activity (contained in the added wheat malt)

An oc-amylase unit according to Sandstedt, Kneen and Blish (SKB) unit corresponds to the amount of enzyme which, at pH 4.7 and 20 ⁰ C. 1 g of soluble starch in the presence of an excess of beta-amylase within one hour so far dextrinized that when the batch solution is colored with iodine, a standardized comparison color results.

The determination is carried out according to the regulation of the ASBC, as it is reproduced in J. de Clerck: Textbook der Brauerei, Vol. II, pp. 597-600, Verlag VLB, Berlin 1965.

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A special feature of the enzyme combination preparation according to the invention is that "when used in raw fruit mash, both the amylolytic breakdown (measured by the increase in fermentable sugars) and the proteolytic breakdown (measured by the increase in pormole nitrogen) and the decrease in viscosity at the same pH Value and same temperature on
fastest run. Tables 1 and 2 use the analytical data determined in 12% wort to show that the mixture given as an example has an optimum pH of approx. 5.0 and an optimum temperature of approx. 58 ^° C. in this regard.

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Table 1

Temperature of the 2nd corn rest (pH 5.0) 50 ° CI 54 ⁰ CI 58 ⁰ C | 62 ⁰ C! 66 ⁰ C

3o i nable iit ml 78 , 5 84 , 2 85 , 5 85 , 0 80 , 0 Vptvnrbarki mg / 100 cp 26th , 81 29 , 6 32 , 0 30th , 2 ' 28 , 2 ? Drmol-N 1 1 , 76 1 , 63 1 , 65 1 , 77 viscosity

Table 2

* PH value at the second . Maiechraat , 5 83 (58 ⁰ O 5 ,8th ml 4.4 4, 7th 5.0 0 31 4th ep 63 1 78 , 9 Apparent 79.2 84 , 2 85 , 0 26th ,8th Fermentability 27.8 30th , 6 32, , 2 1 , 88 ? ormol-N Tig / 100 1.96 1 , 73 1 , 78 Viscosity

409845/0933

Furthermore, the enzyme mixture according to the invention is distinguished by the fact that, through the combination of mutually different, proteolytic enzymes, it contains an optimization effect that has not yet been used in other processes. It has been found that a higher proteolytic "degradation effect in Rohfruchtmaische resulting _s when the total proteolytic activity of the mixture wird- not by a single, but different protease preparations much as possible taught differing specificity of one another particularly good effect showed combinations of plant proteases such as papain , Ficin or bromelin and bacterial proteases, for example protease from B. subtilis.Table 3 uses the example of papain and protease from B. subtilis to illustrate this effect of the increased proteolytic effect used in the enzyme mixture according to the invention by combining individual enzymes with different specificities.

gabeil 3

Activity proportion of papain to bacterial protease in the total protease activity kept constant

100: 0 70:30 30:70 0: 100

Formol-N mg / 100 ml 30.2 32.0 29.8 27.6

Total N mg / 100 ml 116.0 119.1 112.8 107.6

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- ii -

The combination of Fungal proteases such as protease from Aspergillus oryzae, or Aspergillus niger, Aspergillus awamori, Aspergillus usami or Aspergillus saitoi with bacterial proteases such as protease from B. subtilis.

The method according to the invention proceeds in detail as follows: ■

The cereals used (raw fruit) are first finely ground with the addition of 4-20%, preferably 10 % malt (eg wheat darr or wheat green malt) with at least 80 SKB units of oc-amylase per g of dry matter. In this case, it is not necessary to separate off the husks, since in the course of the digestion process according to the invention no high temperatures are used which could have more unpleasant effects in terms of taste and color. On the contrary, the husks remaining in the mash improve the subsequent filtration as an additional filter aid.

After grinding, the mixture is stirred with water, a pH of approx. 5% being established. This corresponds to the optimal value for the first digestion phase. Then, the mash is slowly, ie within 15 to 120, preferably from 45 minutes to a temperature between 67 ° G and 73 ° C, preferably 70 ⁰ C. and 15 to 180 minutes, preferably about 1 hour at this temperature, left (l . Maischrast).

For the initiation of the second digestion phase, the mash must then, if necessary, reach the temperature and pH optimum according to the invention Enzyme mixture can be adjusted. this happens

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by cooling to a temperature of 54 "to 62 ⁰ C, preferably 58 ⁰ C within a period of 10 to 60, preferably 20 minutes and by setting a pH between 4.5 and 5.5, preferably 5.0 by means of acids The enzyme mixture is then added and the set temperature is maintained for 15 to minutes, preferably 60 minutes. This is followed by the usual separation and / or filtration of the mash , which is followed either by the direct further processing of the digestion product obtained as a filtrate or its concentration (eg thickening to syrup, freeze drying or spray drying).

The unusual temperature result of the new mashing process of first about 70 ⁰ C and then about 58 ⁰ C and the taking place only in the second stage of enzyme addition contribute significantly to its exceptional disruption effect and high efficiency. When the raw material is finely ground, the extract yields are 3-6% higher than those of classic mashing processes. Furthermore, at the same time as the good dextrination described above, the viscosity of the mash also decreases rapidly in the initial stage. This enables the processing of highly concentrated mash with a solids content of 20-40%, so that only about half to a third of the mash space of the classic process is required.

The savings made possible by the new mashing technology of oc-amylase has the advantage, besides being cheaper, that lighter seasonings are obtained. Because it was

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found that just technical a-amylase preparations by Oxidase secondary activities cause the wort to darken. A lighter wort color is additionally promoted by the fact that according to the method according to the invention during the first mash rest (at approx. 70 ° C) there is neither significant saccharification nor stronger proteolysis, but the low molecular weight sugars and amino acids only in the second mash rest at a lower temperature are formed. For the dye-forming Maillard reaction are the starting substances (amino acids or reducing Sugar) is therefore only available shortly before the end of mashing and at a lower temperature than usual.

A substantial simplification it also means that -der After the addition of the enzyme, the digestion takes place only in one pH and temperature level. Becomes possible this simplification by the fact that from the large number of enzymes to be obtained from plants and microorganisms specifically ' those are selected which, in terms of pH and temperature, are optimal for raw fruit mash as a substrate with one another to match.

Purely for specific purposes, it is advantageous to bring the digestion product into a stable, concentrated state which also significantly facilitates any transport. For this purpose, the wort is concentrated in vacuo at 40-7O ⁰ C to 40-βΟ ° fo extract content and either used in this syrupy state or ^{spray-dried at 200 to 280 0} C air inlet temperature and 90 to 140 ⁰ C air outlet temperature.

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The spray-drying processes previously known for similar seasonings (see DOS 1 903 400 and DOS 1 918 765) provide for significantly lower drying temperatures. It has been shown in multiple tests that just the higher temperatures used here with regard to the taste of beer made from the wort powder Have advantages. During the drying process, similar roasted products are apparently formed as during the drying process for conventional ones Beers used malt, so that beers from the process geiüäß dried digestion product are produced, in terms of taste are excellent and do not have the "raw fruit taste" that usually adheres to the raw fruit beers. Due to the temperature ranges given above during the drying process, any coloration of the digestion products is possible within the limits specified by the individual beer types. With the lower temperatures, the specified clamping barley seasoning powder, which when re-dissolving in water very light seasonings result with color values, as they are usual for Pilsener worts. With the higher Drying temperatures can be used to produce stronger colored powders, such as those used for beers from Wiener or Munich type are required.

Process example

36 kg of raw fruit (barley, maize, rice, wheat, rye, oats or millet) and 4 kg of wheat lime malt were ground with husks (if present in the cereal in question) to a fine flour content of about 75 ° . The raw materials comminuted in this way were stirred with 100 liters of water at 35-4O ⁰ C in a temperature-controlled mash vessel. The pH of the mash obtained in this way was about 5.8. Within

409845/0933

of 45 minutes, the mash to 7O ⁰ C was heated and "held for 60 minutes to dextrinization and liquefaction at this temperature. After about 20 minutes, cooling to 5S ° C the pH of the mash was with citric acid monohydrate lowered to 5.0 Then 80 g of standardized enzyme combination preparation according to the above specified composition were added. After a 60-minute break in saccharification and proteolysis, the coarse mash fat parts were separated off by decanting. The remaining mash was then mixed with 1 kg of kieselguhr and filtered through a precoat filter. The coagulation and removal of the protein was accomplished by brief heating to 100 ⁰ C, the addition of 0.3 kg kieselguhr and Blankfiltrieren. the resulting as filtrate Auschlußprodukt was then up to an extract content of about 55% concentrated in vacuo at about 55 ⁰ C in the spray tower at 210 ° C air inlet temperature and 95 ° C air outlet temperature with an atomizing pressure of 2.5 atmospheres (Two-substance nozzle) dried.

After redissolving the process according to prepared barley seasoning powder this was boiled with hops for 90 minutes and fermented into beer in the usual way, the smell pure in the drink pleasantly full-bodied, pure, soft and the bitterness was judged to be very mild. Particularly noteworthy is the excellent one Stability of the beer produced from our pulverized digestion product, which is also shown in Table 4 for Expression comes.

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Table 4

Apparent extract,% -. 1.60

Real extract, % 3.36

Alcohol,% 3.88

Original wort, % , 10.92

Discharge degree of fermentation apparent , #. . 85.4

Final degree of fermentation apparent,% 85.5

PH value .. . . ,. ... . . 4.5

color, EBC. . . 11.5

Foam Number (Ross & Clark) 144

Bitter substances, EBC. 2-0.6

Tannins, mg / 1 ...... 93

Antho cyano gene, mg / 1 .......... .... 19th

Total N, mg / 100 ml 99.7

Coagul. N, mg / 100 ml. ...... 1.8

High mol. N, mg / 100 ml. 33.6

Middle mol. N, mg / 100 ml,. . 23.9

Niedermol. N, m ^ / 100 ml 42.1

Formol-N, mg / 10O ml 14.5

Cold haze according to EBC .

- 24 h at O ⁰ C 0.0

48 h at 0 ° C .. 0.3

1 warm day at 40 ⁰ C 0.5

Warm days up to 2 EBC turbidity units' -.

reached (40 ° / 0 ° / 40 °) ... ... 9.5

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Claims (1)

Claims Q). Process for obtaining a digestion product from Cereals with the addition of malt and enzymes, thereby Inet that the crushed raw fruit with ί üis 20 j'o malt is made into a mash, this · ■.;: a temperature between 67 C and 73 C and utv / a Ib for 1 iO minutes kept at this temperature ··· .jrd, then to 54 ⁰ C to 62 ⁰ C is cooled and a pH value of 4.5 to 5.5 is set, whereupon an enzyme mixture containing an aryloglucosidase and at least one microbial and / or vegetable protease "is added and the set temperature is maintained for 15 to 300 minutes, before the customary separation and / or filtration and, if desired, the concentration and / or drying of the digestion product obtained as piltrate takes place. 2. The method a ^ ch claim 1, characterized in that as Cereali-jii raw fruit barley, rye, wheat, maize: veis, oats or millet each alone or in any mixture - "i.jigesetzt". 5. The method according to claim 1, characterized in that per kg of cereals used 1000 to 3000 amyloglucosidase units and 100 to 500 protease units be added in the form of an enzyme mixture. 409845/0933 Ai method according to claims 1 and 3 ». Characterized in that the added protease activity originates from at least two different sources. 5. The method according to claim 1, 3 and 4 ». characterized, that the protease activity originates on the one hand from plants or fungi and on the other hand from bacteria. 6. The method according to claim 1, 3 and 4, characterized in that the added protease activity comes to about 70 i ° from plant protease .and to about 30 % from bacterial protease. 7. The method according to claim 1, characterized in that the mash has a pesticide content of -20 to $ 40> having. S. The method according to claim 1, characterized in that the malt used has an activity of at least 80 SKB units per g of dry matter. 9. The method according to claim 1 and 8, characterized in that that used as mals wheat lump malt or wheat green malt will. 10. The method according to claim 1, characterized in that the digestion product obtained as filtrate is concentrated in vacuo to 40 to 70 fi extract content. 11. a-amylase-free enzyme preparation for the digestion of cereals, characterized in that it contains amyloglucosidase and at least one microbial and / or vegetable protease contains. . / 19th 409845/0933 12. Enzyme preparation according to claim 11, characterized in that that its protease activity comes from two different sources. 13. Enzyme preparation according to claim 11 and 12, characterized in that that the protease activity originates on the one hand from plants or fungi and on the other hand from bacteria. . A vin preparation according to claim 11, characterized in that the protease activity originates from about 70% plant protease and about 30 % from bacterial protease. 15. Enzyme preparation according to claim 11, characterized in that there is an activity ratio of the amylogiucosidase units to protease units from 1 to 0.03 "to 0.5. ok Enzyme preparation according to Claims 11 and 15, characterized in that that there is an activity of 500 to 1500, preferably 1000 amyloglucosidase units per g and 50 to 250, preferably 100 protease units per g and with an erizyme-free substance to these activity values is standardized. 17. Enzyme preparation according to claim 10, 15 or 16, characterized in that that in addition to amylogiucosidase and at least one microbial and / or vegetable protease Contains cellulases, hemicellulases and / or pectinases. / 20 409845/0933 18, enzyme preparation according to one of claims 11 to 17, characterized in that when it is used in raw fruit mash, the optimum of the proteolytic effect, measured by the increase in pormol nitrogen, the optimum of the amylolytic effect, measured by the increase in fermentable sugars and the optimum of no further than, preferably not more than apart viscosity reducing effect by 0.6 pH-values and 8 ⁰ C by 0.3 pH and 4 ° C. 19. Enzyme preparation according to claim 18, characterized in that da3 the pH optimum of its enzymes is between 4.7 and 5.3, preferably between 4.9 and 5.2 and the temperature optimum between lies. between 54 and 62 ⁰ G, preferably between 56 and 60 ⁰ C 20. Production of an enzyme preparation according to one of claims 11 to 19, characterized in that amyloglucosidase mixed with at least one microbial or vegetable protease, if desired also cellulases, hemicellulases and / or pectinases and standardized by mixing with an enzyme-free substance. 21. Use of an enzyme preparation according to one of claims 11 to 2.0 for the digestion of raw cereal fruit. 22. The method according to claim 1 and 21, characterized in that the digestion product is dried in a spray dryer will. / 21 409 845/093 23. The method according to claim 22, characterized in that the spray-drying an air outlet temperature of 90 to 140 ⁰ G takes place at an air inlet temperature of 200 to 24O ° and G. 24. Spray-dried digestion product made from raw cereal fruit, characterized in that it was obtained according to the method of claim 23. 25. Use of the digestion product according to one of claims 1 to 9 or 24 as a base for brewing purposes, others Alcoholic or non-alcoholic beverages, malt sweets and other confectionery, microbiological nutrient media, dietetic and children's food, baking additives, etc. 409845/0933