DE4116744A1 - Seasoning prodn. from protein-rich plant raw material - by addn. of protease(s) to finely distributed substrate in water and hydrolysis to specific amino-nitrogen-total nitrogen ratio, useful as foodstuff - Google Patents

Abstract

Prodn. of enzymatic seasoning or seasoning from plant raw material where the substrate is finely distributed in water is hydrolysed by (i) proteases to an amino-nitrogen/total nitrogen ratio of 0.5 or above. Pref., (ii) peptidases; (iii) glutaminases; (iv) maceration enzymes, i.e. cellulases, pectinases, or amylases are also added either at once or gradually by first adding (i)+(ii)+(iv) and then (iii), or (i)+(ii) and then (iii)+(iv). Pref., the hydrolysis of protein rich raw material e.g. soya flour, soya coarse meal or wheat gluten is at a pH above 7. USE/ADVANTAGE - A seasoning for foodstuff is provided with an amino-nitrogen/total nitrogen ratio of at least 0.5 which has a high content of dry substance and glutamic acid without a bitter taste.

Description

The invention relates to a method for enzymatic Wür Manufacture of ze or spices from vegetable raw materials.

Seasonings are obtained by hydrolysis of protein substances Products used as flavoring food additives will; they arrive as liquid food seasonings, as one steamed and concentrated wort paste or dried as Flavoring powder in the trade.

Seasonings increase the individual taste of each Food. This is achieved by hydrolysis of amino acids obtained from vegetable protein - in the we considerable glutamic acid. The flavor of the spice are u. a. sulfur-containing amino acids and lactones. For hydrolysis mainly serves an inorganic, strong acid, in which Usually hydrochloric acid. Sulfuric acid is used when Salt-free seasonings are to be produced. In the Hydrolysis with an inorganic acid becomes ratios from amino nitrogen to total nitrogen of about 0.8 enough. This gives information about the degree of degradation of the Proteins. In fermentatively produced wort, e.g. B. Soy sauce, is a ratio of amino nitrogen to total nitrogen reached 0.5. Protein hydrolysis with En Zymen is known, but these enzymes have been overcome used to modify proteins, d. H. only for Dismantle part to add other functional properties cause, because in the enzymatic hydrolysis in technical  Scale not the desired typical flavoring substances, son which often produce bitter peptides.

The invention has for its object a method for enzymatic wort production from vegetable raw materials to create a relationship with amino stick achieve total nitrogen of at least 0.5 leaves, but with no or only very little bitters peptides are formed and this leads to a flavor that the contains the desired flavorings.

Based on this task, a ver driving the type mentioned suggested that invent according to a protea finely divided substrate in water sen added and the substrate using the proteases under mild conditions is hydrolyzed until a ratio from amino nitrogen to total nitrogen of about 0.5 or more is achieved. Advantageously, in addition to The proteases exopeptidases are used to make a wide re splitting the peptides obtained into amino acids to reach.

If glutaminases are also used, Desirably released from enzymatic glutamic acid tem glutamine gained what the proportion of from the proteins obtained glutamic acid increases.

To also get the starch, cellulose and other carbohydrates cleave, which contributes to the formation of the typical wort aroma carries additional macerating enzymes, such as cellula sen, pectinases, glucanases and amylases can be used.

The various enzymes can be processed in a single step drive in a mixture, but it is preferable  pull, group the enzymes in a multi-step process to be used wisely.

The glutaminase in particular demands for its optimal Activity has a pH above 6.5. With the enzymatic However, hydrolysis using proteases and exo-peptidases decreases the pH continuously as a result of the release of Acid groups of the amino acids. The steady drop in pH who tes is an indicator of the persistence of the reaction so that the end of the reaction is reached when the pH value remains constant. Now the pH can be adjusted by adding Alkali such as caustic soda can be raised to over 7. After that can be glutaminase alone or in a mixture with the macerie enzymes in the second stage of the process be so that in the course of the reaction in the second Stage an increase in the amino nitrogen content, the content of glutamic acid and the dry matter.

A distinction can be made in the method according to the invention Use high protein vegetable raw materials. The Use of soy flour and / or ground soybean meal and Wheat glue results in particularly high-quality condiments. As a plant Protein carriers can also include corn glue, rice glue and Serve peanut meal.

In detail, the process can be carried out in the following steps expire:

Soy flour and wheat gluten are mixed in one in cold water Dry matter content of about 20% mashed. Then ßend is heated to 120 ° C for one hour. After that is ver with cold water to 10% dry matter content thins and the enzymes are in a subset of the reaction approach mashed. Enzymation takes place at 40 ° to  42 ° C for about 12 hours with stirring. The sinks pH steadily decreases to 7.5 at the end of the first reaction is raised. Another enzymation follows at 40 ° to 42 ° C for about 4 hours with stirring. After that the non-hydrolyzed solids can be separated, the liquid is mixed with salt, concentrated and by heat at at least 80 ° C for 20 minutes fourth.

With regard to wort, further raw materials can be added to the substrate such as yeast, yeast extract, vegetable extracts, spices and / or Ge seasoning extracts and also flavoring with regard to seasonings amplifiers, flavors, sugar, and color-containing plants liche processing products are added, these Addition from the beginning when mashing soy flour and Wheat glue and / or after filtering and / or after the deactivation can take place. As enzymes can be in Mixture in equal parts by weight 1. Protea se, 2nd protease, 3rd protease and 4th protease as well as the white tere amylase, glutaminase and cellulase to each other same weight, possibly different for the first mixture divide and by 5th and 6th protease in liquid suspended Use shape.

A proven mixture can consist of 50 mg / l 1st to 4th Pro tease and 0.1 ml / l 5th protease and / or 6th protease in the first enzymation and a mixture of 50 mg / l each Glutaminase, cellulase, amylase and from 0.1 ml / l glucoamy read in the second enzymation for a substrate from soy flour and wheat glue in a ratio of 80:20 the.

For a substrate made of soy flour and wheat glue in the ratio nis 60:40, a mixture of 50 can be conveniently  mg / l 1st to 4th protease, 5th protease and 6th protease in the first enzymation and a mixture of 50 mg / l glutamina se, amylase and cellulase in the second enzymation put.

When preparing the substrate it should be noted that at the classic fermentation organisms through the substrate grow and their enzymes are finely divided at all points place in the substrate. In contrast, when using isolated enzymes only the surfaces of the substrate grains reached. It is therefore necessary to use the Sub punch evenly and finely ground.

To prepare the reaction batch, the are finely ground mixed substrates and suspended in water. To Ver Avoiding lumps is only subsequently heated and the batch was held at 120 ° C. for about an hour. Here is achieved by that inhibitors for the proteases zer be disturbed. Furthermore, the substrate is opened sen, and the first proteins go into solution. A part of Protein is denatured, making it easier Proteases access to the substrate. Also the substrate sterilized what for the subsequent long litigation is required under mild conditions because mesophilic Organisms otherwise during the enzymatic conversion would grow and destroy the approach. The substrates can mixed in any composition and the other Undergo treatment. In preparation, the Appropriately double concentrated with one Ge hold heated to 20% dry matter, then to react and to lower the temperature to 10% dry sub punch content to be diluted. The enzymes can be found in the commercially available forms are used.  

The enzymes are weighed in and with a small amount of the reaction mixture mashed. After that they become the tempered approach added.

The first enzymation with the mixture of the protein begins ases 1 to 6. The pH value falls as a result of the hydrolysis reaction continuously due to the release of acid groups. The steady drop in pH is an indicator of the on take the reaction. The first enzymes are used in primarily from the proteins amino acids and peptides released. Since it is desirable to have as large a menu as possible gene to release glutamic acid, some of which pri is released from the proteins, no less However, portion made from glutamine by deamination a second enzymation is carried out. The se second enzymation takes place by adding glutaminase, after the pH of the batch after finishing the first Enzyme treatment with sodium hydroxide solution to a value above 7 was discontinued.

The second enzymation using glutaminase can otherwise only run effectively when sufficient quantities are available of glutamine are present. This glutamine is also created during the first enzymation. After weighing the Enzy For the second enzymation, these enzymes are converted into one small amount of the reaction mixture mashed and in the Approach mixed.

The second enzymation is also at 40 ° to 42 ° C carried out and takes 4 hours with stirring. Preferred two For the second enzymation, not only are glutaminase, but also macerating enzymes such as amylases, cellulases and glucanases added. After finishing the second  Enzymation, the reaction mixture is filtered, the won Dilute basic wort by adding table salt lized, concentrated in vacuo and by heat at inactivated at least 80 ° C for 20 minutes. As white Other additives can be spices, spice extracts, herbal extracts ge, yeast and yeast extracts as well as appropriate Sugar, vegetable processing products containing colorants, Flavor enhancers and flavors are typed in both in the first reaction batch before the first Enzymes as well after the second enzymes or also add after deactivation.

Hydrochloric acid and baking soda can be used to adjust the pH serve lye.

For the enzymatic wort production from soy flour and white zenkleber in a ratio of 80:20 has been chosen for the first Enzyme a mixture of 50 mg / l 1st to 4th protease together with 0.1 ml / l 6th protease and a mixture for the second enzymation of 50 mg / l glutaminase, cellulase and amylase as well as 0.1 ml / l glucoamylase.

For the enzymatic wort production from soy flour and white zenkleber in a ratio of 60:40, however, has in one one-step enzyme process a mixture of 100 mg / l 1st to 4th protease, 300 µl / l 5th protease, 50 mg / l Glu taminase, 100 mg / l amylase and 50 mg / l cellulase proven.

It also emerged from tests that the enzyma tables wort production from soy flour and wheat glue in Ratio 60:40 in a two step process a Ge mix from 50 mg / l 1st to 4th protease and 300 µl / l 6th pro tease for the first enzymation and a mixture of each  50 mg / l glutaminase, amylase and cellulase for the second en Zymierung also gave good results.

While the proteases in the acidic range are optimal and with work well, the exopeptidases are on one slightly alkaline range. To optimize the pH of all enzymes used in the course of the process run, the pH of the reaction mixture is not con kept constant, but automatic pH lowering left influenced. It turns out that the use of the 4th to 6th proteases and a degree of hydrolysis of more than 30% determined bitter taste with the be written approaches of the inventive method ent fell.

Regardless of the purity of the enzymes, the products are based on the following activities:
1. protease - predominantly alkaline endoprotease,
2. Protease - predominantly neutral endoprotease,
3. protease - predominantly acidic endoprotease,
4. Protease - predominantly neutral exoprotease,
5. Protease - alkaline exoprotease and endoprotease,
6. Protease - predominantly alkaline exoprotease.

The wort produced according to the invention contains 3% total nitrogen in a ratio of amino nitrogen to total nitrogen of at least 0.5. The wort has a high Content of dry matter and glutamic acid, has none Bitter taste and has the desired flavor flavor. Seasonings with a total nitrogen can also be used produce content of 1% and intermediate values.

Claims (18)

1. Process for the enzymatic wort or condiment production from vegetable raw materials, in which

• - Proteases added to a finely divided substrate in water and
• - The substrate is hydrolyzed by means of the proteases until a ratio of amino nitrogen to total nitrogen of 0.5 or more is reached.

2. The method according to claim 1, characterized by the additional use of peptidases for further up cleavage of the peptides obtained into amino acids. 3. The method according to claim 1 or 2, characterized by the additional use of glutaminases for free setting of glutamic acid from enzymatically released Glutamine. 4. The method according to one or more of claims 1 to 3, characterized by the additional use of macerating enzymes such as cellulases, pectinases and Amylases. 5. The method according to one or more of claims 1 to 4, characterized in that the enzymes in one stage process can be used in a mixture. 6. The method according to one or more of claims 1 to 4, characterized in that the enzymes in a more stage procedure can be used in succession.   7. The method according to one or more of claims 1 to 4, characterized in that in one stage the Pro teases, peptidases and macerating enzymes in a mixture and used glutaminase in a second stage will. 8. The method according to one or more of claims 1 to 4, characterized in that in a first stage the proteases and peptidases and in a second stage the macerating enzymes and the glutaminase in a mixture be used. 9. The method according to one or more of claims 2 to 8, characterized in that the pH of the solution to optimize the action of the peptidases and / or Glutaminases after exposure to proteases and / or macerating enzymes to a value of more than 7 is set. 10. The method according to one or more of claims 1 to 9, characterized by the use of difference protein-rich vegetable raw materials. 11. The method according to claim 10, characterized by the Use of soy flour and / or finely ground soy grist and wheat glue. 12. The method according to claim 10 or 11, characterized by the following steps

• - Mashing soy flour and wheat gluten in water to a dry matter content of about 20%,
• - heating to 120 ° C for one hour,
• - Dilute with cold water to 10% dry matter,
• Mashing the enzymes into a cooled portion of the reaction mixture,
• - Enzymation at 40 ° to 42 ° C for about 12 hours with stirring,
• - raising the pH to 7.5,
• - Enzymation at 40 ° to 42 ° C for about 4 hours with stirring,
• - Split off,
• - concentration,
• - Inactivate by heat at at least 80 ° C for 20 minutes.

13. The method according to claim 12, characterized by the Addition of further raw materials such as yeast, yeast extract herbal extracts, spices, spice extracts and Sugars, flavor enhancers, flavors and coloring containing vegetable processing products. 14. The method according to claim 13, characterized in that the other raw materials when mashing the soy flour and wheat glue and / or before inactivating be added. 15. The method according to claim 12, 13 or 14, characterized records that after raising the pH to 7.5 a second enzymation by adding peptidases and / or glutaminases is introduced. 16. The method according to one or more of claims 1 to 15, characterized by the use of a mixture from 1st to 4th protease with identical Ge important parts, glutaminase, amylase and cellulase in un  equal to each other, if necessary different from the first mixture those parts by weight and of 5. protease and / or 6. Protease in liquid suspended form. 17. The method according to claim 16, characterized by the Use a mixture of 50 mg / l 1st to 4th Pro tease and 0.1 ml / l 5th and 6th protease in the first enzymation and a mixture of 50 mg / l each Glutaminase, cellulase, amylase and 0.1 ml / l Glu coamylase in the second enzymation for a substrate from soy flour and wheat gluten in a ratio of 80:20. 18. The method according to claim 16, characterized by the Use a mixture of 50 mg / l of Prozym 6, 1. to 4th protease and 0.3 ml / l 5th protease and / or 6. Protease in the first enzyme and a Ge mixes of 50 mg / l glutaminase, amylase and cellu read out in the second enzymation for a substrate Soy flour and wheat gluten in a ratio of 60:40.