FI79858C - FOERFARANDE FOER EXTRAHERING AV ENZYMER UR SPANNMAOL. - Google Patents

Description

1 79858

METHOD FOR EXTRACTING ENZYMES FROM CEREALS - FÖRFARANDE FÖR EXTRAHERING ENZYMER UR SPANNMÄL

The invention relates to a process for extracting enzymes from cereals and / or cereal-derived material, wherein the cereals and / or cereal-derived material are extracted and the enzyme is separated from the extraction solution.

The invention relates to a process for the extraction of any enzyme contained in and extractable from cereals from any cereals and / or cereal-derived material, in particular for the extraction of β-amylase from barley.

β-amylase is a starch hydrolyzing enzyme that cleaves the 1,4-bonds of starch from the non-reducing ends of glucose chains. The result is maltose or maltose and maltotriose, depending on the origin of the β-amylase. If the β-amylase is derived from cereals, the hydrolysis results mainly in maltose. Starch 20 hydrolyzed by β-amylase from cereals is suitable e.g. as a raw material for beer or for use in the confectionery industry, β-amylase is of great importance in the brewing industry.

β-amylase is formed in and stored in grains during growth, β-amylase is located in the grain relatively close to the shell, in the subaleuron layer of the grain. To extract β-amylase, the grain is usually ground or ground before extraction, or β-amylase is extracted from mechanically separated parts of the grain, such as bran. The extraction usually uses water or a suitable buffer solution, the β-amylase can be separated from the extraction solution by concentrating it after separation of the precipitate using ultrafiltration or evaporation. In addition, it is known to use a sulfur compound, such as SO 2, in the extraction to release β-amylase from other proteins.

It is known from German patent publication DE 1 792 362 to extract β-amylase from wheat bran with water at a temperature of 20 to 40 ° C, the amount of water being four times the number of bran 2 79858. The extraction time was lr5 h.

It is known from Finnish patent publication FI-61516 to extract β-amylase from whole or partially peeled barley grains in the operation of the grain's own membranes as a kind of filter. In this case, the extraction temperature was 20 to 50 ° C, the extraction time was more than 5 h. Methods of the same type are known from U.S. Pat. No. 3,492,203 and U.S. Pat. No. 4,675,296.

Other methods for separating α- and β-amylase are known from SE 372 022 and US 2,594,356.

There are drawbacks to known methods for extracting enzymes from cereals and / or cereal-derived material. The raw material, i.e. ground, crushed or otherwise treated or whole grain, is accompanied by microorganisms which, for example at a temperature of 20-50 ° C which is favorable for them and utilizes the nutrients derived from the grain, multiply strongly, especially when the extraction time is relatively long , e.g., 1.5 to 5 h. In particular, an increase in lactobacilli causes an increase in acidity, which in turn leads to inactivation of β-amylase. By adding a base, an excessive increase in acidity can be prevented, but the base must be added in such amounts that evaporation becomes difficult and the product saline. Maternal birth naturally raises the cost.

The extraction solution obtained from barley extract contains soluble constituents of cereals, some sugars and starch and a moderate amount of protein. From the extraction solution, an enzyme extract containing β-amylase and an extraction solution suitable for animal feed can be prepared by evaporating water as described above. However, the evaporation of water consumes energy, and on the other hand, water must be constantly added to the extract. Circulation of the extraction solution increases the contamination problem-3 5 land.

The object of the present invention is to eliminate the above-mentioned problems.

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In particular, it is an object of the invention to provide a method by which the problem of contamination can be kept under control and by which the inactivation of β-amylase can be prevented.

It is a further object of the invention to provide a method by which the extraction of β-amylase from barley can be carried out as a continuous and substantially closed system by circulating the extraction solution.

With respect to the features of the invention, reference is made to the claims.

According to the invention, the grain extraction is carried out with an ethanolic solution of water, ethanol concentration 6-11% by weight. Such ethanol concentrations have been found to control microbial activity. When β-amylase is extracted and the ethanol concentration is above 11% by weight, β-amylase precipitates. At an ethanol concentration of less than 6% by weight, the contamination problem remains uncontrolled. The fact that the contaminants remain under control even at an ethanol concentration of 6% by weight is surprising, since, for example, mild wines have to be sulfurized to control 20 microbes, even if they contain 9-11% ethanol.

The addition of ethanol according to the invention reduces the viscosity of the aqueous solution and increases the differences in density, which significantly improves the separation of solids from the solution. The precipitate containing starch and proteins can be better separated from the extraction solution. Ethanol also precipitates non-specific protein, leaving it round with the precipitate. The quality of the enzyme extract is improved and its further purification is substantially facilitated. The addition of ethanol according to the invention makes it possible to recycle the extraction solution, e.g. by dividing the solution and separating the part to concentrate the β-amylase, the part being returned directly to the extraction.

Concentration of the enzyme can be performed by any method known per se, such as ultrafiltration or evaporation. When using evaporation, the mixture of evaporated and condensed water and ethanol can be returned to * 79858 extract.

Thanks to the invention, the β-amylase is better recovered, the β-amylase is enriched in the extraction solution, which facilitates the extraction process. The extraction step can then be shortened so that its duration is only about half an hour.

The extraction solution and the water obtained from it during the separation and the energy with the water can be saved. Also, a substantial portion of the ethanol added to the leach solution to control microbial activity can be kept in circulation. Only the amount of ethanol consumed when the precipitate is separated from the extraction solution is consumed.

The invention is suitable for extracting any water- and ethanol-soluble enzyme from cereals or cereal-derived material. In this context, cereal or cereal-derived material means any cereal such as barley, wheat, oats, rye, maize, etc. and, accordingly, any cereal-derived material such as cereal husks, any part of the cereal grain 20 or otherwise a material made from cereals such as ground, crushed or otherwise comminuted grain-derived material or other cereal-derived material made from cereals by any process.

If desired, other additives may be added to the extraction solution, e.g. to enhance the extraction of the enzyme. In this case, for example, the addition of sulfur dioxide comes into question.

The temperature used in the process according to the invention is e.g. 20 to 60 ° C, suitably 30 to 55 ° C, e.g. about 45 to 55 ° C.

The process can preferably be carried out as a continuous process, as is known in the art. In this case, a part of the extraction solution is continuously discharged, from which, on the other hand, the enzyme is separated, e.g. by said evaporation, filtration or the like separation process, and from which the separated extraction solution can be advantageously returned to the extraction cycle. In this case, of course, cereals or cereal-derived material, as well as water, ethanol or another substance used in the extraction solution, are continuously added to the extract; correspondingly, 5 precipitates are continuously removed from the extract.

If desired, the method can also be implemented as a batch process. Even then, any desired additive can be added to the extraction solution. The batch process can be carried out in a manner known per se.

The invention will now be described in detail by way of exemplary embodiments, which, however, are intended only to illustrate the invention.

Example 1;

In one continuous process, circulating process 15 was continuously fed with ground grain, ethanol and sulfur dioxide, as well as water. The precipitate was continuously removed from the circulation process. The extraction solution was divided into two portions, one was concentrated and the other was recycled; the enzyme product was separated from the portion of the extraction solution passed to the concentration, and the resulting solution was recycled. The mass flows / unit of time were as follows: feed 1000 kg / h grain, water feed 650 kg / h, sludge removal 1600 kg / h, recovery extract returned to the process 1500 kg / h, concentration extract extracted 550 kg / h, separated enzyme product β-amylase 50 kg / h, the separated extraction solution returned from the concentration to the process is 500 kg / h. Ethanol was added so that the ethanol content of the circulating solution was 8% by weight on average. The residence time in the process was 0.5 h, temperature 50 * C.

30 Example 2:

In another process, in a circulating process such as Example 1, ground barley was slurried in water at 1000 kg / h, water was used at 2000 kg / h, water was recycled water after precipitation, to which ethanolic water from the concentration of the extract was added. Approximately 25 kg of ethanol / h and 1.5 kg of Na metabisulfite / h were also fed to the extract. The average residence time 79858 was 30 min, temperature 50 ° C. The ethanol content was about 8% by weight.

The precipitate was removed by decantation and a separator. Pure water 650 5 kg / h was fed to the precipitate to remove the precipitate. The precipitate was about 1600 kg / h. The β-amylase-containing extract from the precipitate was partially recycled back to the extraction step, 550 kg of extraction solution / h was separated for evaporation. Evaporation gave 50 kg / h of β-amylase-containing syrup and 500 kg / h of ethanol-containing condensate, which was recycled to the extract.

The activity of the obtained β-amylase was 600-800 ° L / ml.

The process was continued for 48 h.

Claims (5)

A method for extracting enzymes from semen and / or semen source material, wherein semen and / or semen source material is extracted and the enzymes are separated from the extraction solution, characterized in that the enzyme extracted is B amylase. and that as an extraction solution aqueous ethanol of 6-11% by weight is used. 10 2. Process according to claim 1, characterized in that the extraction solution contains ethanol 7-9% by weight. Process according to claim 1 or 2, characterized in that the extraction is carried out in a continuous circuit process, in which in the process continuously seed and / or material of cereal origin, water and ethanol are fed, in the process the precipitate and enzyme product are removed and the enzyme product is removed. is separated from the extraction solution, which is returned to the process after the separation of the enzyme product. Method according to any of claims 1-3, characterized in that the temperature of the circuit process is 20 - 60 ° C, preferably 30 - 55 ° C, preferably 45 - 55 ° C. 25 Process according to any of claims 1-4, characterized in that the average residence time in the process is 10 minutes - 2 hours, preferably 20 minutes - 1 hour.