LT4757B - MULTIENZYM COMPOSITION, METHOD OF RECEIVING IT AND USING BREAD PRODUCTS TO IMPROVE - Google Patents

Abstract

The invention concerns the food industry and can be used in bread production. The aim of the invention is to improve pastry products using micro-organism products of natural origin. The aim is achieved by using multienzyme composition with ferment activations: alpha-amylase, glucoamylase and xylanase, their size unit/g is as follows:alpha-amylase: 1-1,000,glucoamylase: 1-1,000,xylanase: 1-1,000.The composition is made by mixing fluid or dried filtrates, breeding different fungi with micro-organism stem belonging to Trichoderma and Aspergilus genus using the depth method. Trichoderma genus fungus is T. viridae or T. reesei, or T. longibranchiatum. Aspergilus genus fungus is A. awamori, A. niger or A orryzae. The best performance is achieved when the proportion of alpha-amylase and glucoamylase is 1:1-1:10. Multienzyme composition is used for enlarging the volume of the bread, improving the structure of the crumbs and the appearance of the crust and slowing the perishing process.

Description

The invention relates to the food industry and can be used in the manufacture of bread products.

When making bread products with yeast, especially white bread, it is very important to get high volume products. Equally important are the appearance, crust color, pulp structure and aging resistance of these baked goods. These properties of bakery products depend on many factors, most of which are related to the properties of the flour used to make the dough. However, even with the highest quality flour, certain additives can improve the properties of baked goods. The use of inferior flour improves the improving properties of such additives.

Various chemical additives (emulsifiers, oxidizing agents, reducing compounds, pH stabilizers, mucins, fat) are known to improve flour, dough and bakery products (Sauvarin, S.P., Young L.S., Technology of Breadmaking, London: 1998). However, many of the chemical dough enhancers used cause significant side effects, such as unpleasant organoleptic effects. In addition, chemical enhancers such as bromates are banned in many countries.

The most commonly used dough and bakery products are enzymes of microbiological origin (such as α-amylases, proteases, pentozonases, lipases, oxidases, etc.) which exist in one form or another in the human body.

The use of the microbial α-amylase in the bread industry is mentioned in several patents. For example, fungal amylase and fungal protease, which are used in combination with L-cysteine and ascorbic acid, are known to improve the quality of white bread (U.S. Pat. No. 3,934,040).

The fungal enzymes α-amylase, glucoamylase and lipase used in combination with chemical compounds are known to slow down the baking of breadcakes (U.S. Pat. No. 4,160,848). The aLT 4757 B amylase is emphasized as the major enzyme. The fungal α-amylase produces fermentable sugar by breaking down the damaged starch grains in the flour. In addition, dextrins are formed, which can improve gas retention in the dough, resulting in better dough rise.

A known multienzyme composition containing xylanase, glucosidase and glucose is used to improve bakery products (EP 396162).

The closest known multienzyme composition used to improve dough and bakery products, increase their volume and inhibit ringing is composed of enzymes belonging to different classes such as carboxidases and lipases (Patent WO 95/23515). The said patent describes the use of xylanases from Aspergillus aculeatus in the manufacture of bakery products. However, the use of xylanase is associated with some undesirable effects, such as the dough becoming too sticky. It is noted that although xylanase may be used as a single enzyme, its combination with other enzymes may be more effective. Possible additional enzymes include α-amylase, glucoamylase, protease, peroxidase, lipase, peptidase, maltogenase. Assays with α-amylase, lipase and oxidase are presented.

The object of the invention is to improve pasta products with products of microorganisms of natural origin.

This object is achieved by a multienzyme composition having amylase, glucoamylase and xylanase enzymatic activities with AV / g activity:

α-amylase 1-1000, glucoamyase 1-10000, xylanase 1-10000.

The multienzyme composition is obtained by mixing the class of enzymes carbohydrase α-amylase (EC 3.2.1.1), glucoamylase (EC 3.2.1.3) and xylanase (EC 3.2.1.8). All these enzymes are of fungal origin, but are obtained by culturing different strains of microorganisms (micromycetes). This provides a broad spectrum of escorting enzymes (cellulases, hemicellulases, pectinases, glucanases) which further improve the properties of the multienzyme composition. Thus, such an enzyme mixture is superior to an enzyme mixture. it is obtained by the direct strain of one micro-organism. Enzymes are obtained from standard micromycetes used in the biotechnology industry under various manufacturing conditions. However, the conditions and quality of production of these enzymes must meet the requirements of the enzymes used in the production of the food. The microorganisms used to produce the enzymes may belong to the genera Aspergillus and / or Trichoderma. Micromycetes of the genus Aspergillus can be, for example, A.niger, A. oryzae, A. awamori, etc. Micromycetes of the genus Trichoderma may be T. reesei, T. viride, T. longibranchiatum and others. liquid or dried culture liquid ultrafiltrates of cultured micromycetes A. awamori and T. reesei. In addition, the fermentation conditions are selected so that a certain ratio of α-amylase to glucoamylase is present in the culture fluid. When using accelerated bread baking technology (without intermediate dough maturation), the optimal ratio of α-amylase to glucoamylase is 1: 1 to 1:10. If classic bread baking technology is used, the optimal amount of α-amylase in the multienzyme composition may exceed that of glucoamylase.

The proposed multienzyme composition is used to improve bread bakery products: to increase volume, to improve appearance, to slow down aging.

The activities of enzymes in a multienzyme composition are defined as follows.

A unit of amylase activity (AAV) is defined as the amount of enzyme which is capable of degradation under standard conditions (30 ° C, pH = 6.0, 10 min) to 1 g of starch of various molecular weights, which is 30% of the starch involved in the reaction.

The unit of glucoamylase activity (GAV) is defined as the amount of enzyme which liberates 1 µmol of glucose per minute at a temperature of 30 ° C and a pH of 4,7.

The unit of xylanase activity (CAV) is the amount of enzyme which, under standard conditions (50 ° C, pH = 5.0, 10 min), is capable of liberating 1 µmol of reducing sugars (xylose) per minute under the action of xylan.

The proposed multienzyme composition may contain 1-1000 AAV / g aamylase, 1-10000 GAV / g glucoamylase, and 1-10000 KAV / g xylanase. Best results are obtained when the amount of α-amylase in the multienzyme composition is 200 to 500 AAV / g, the amount of glucoamylase is 1 to 3000 GAV / g and the amount of xylanase is 1000 to 10,000 KAV / g.

The multienzyme composition may be in any form suitable for use and must meet the quality requirements for enzymes used in the food industry. For example, it may be in the form of a dry powder, granular or liquid because its active ingredient is highly soluble in water. Dry powders or granules are more suitable when the multienzyme composition is to be added to the flour or dough together with other dry components, while the liquid form is more suitable when the multienzyme composition is to be used in combination with liquids.

The multienzyme composition may be used in pure form or may be standardized (up to certain enzymatic activities) in flour, glucose or milk powder. If the multienzyme composition is used to improve the flour, it is best initially to prepare a mixture of 1 part multienzyme composition and 9 parts flour. This mixture is then added to the flour.

The multienzyme composition can be used as a standalone enhancer in flour, dough and / or bread products, or in combination with any other additives used in the industry, such as ascorbic acid, cysteine, bromine, purifiers, etc., as well as with any other enzymes, e.g. glucosoxidase, lipase, phospholipase and the like.

In most cases, the dosage of the multienzyme composition will range from 0.01% to 0.5% by weight of the added flour, preferably from 0.01% to 0.1%. There is no linear relationship between the amount of enzymes used and the quality of the bread to be baked. The latter also largely depends on the quality of the flour. Therefore, the exact amount of multienzyme composition used can only be determined experimentally.

Examples of use of the invention.

Test 1. Use of a liquid multienzyme composition for baking white bread

The test bakes were made with flour of the following parameter: humidity - 13.2%, fall number - 372 s, gluten content - 28.9%, gluten index - 96 units, water [beverage 14% moisture - 57.2%, dough forming time - 2.25 minutes, dough stability - 4.5 minutes, dough dilution degree - 65 FV, farinograph quality number - 75. Dough composition: 550 C wheat flour - 100 kg, dry yeast - 0.7 kg, salt - 0.6 kg, water. Bread in the first (control) group was baked from such dough without any enzyme additive in the test. A second group of baked goods was obtained from said dough; with the addition of 0.2 ml / kg of the proposed enzyme composition in which sterile ultrafiltrates of Aspergilus awamori and Trichoderma viride culture fluid were taken in a 20:20 ratio. A third group of baked goods was obtained from said dough by the addition of 0.35 mi / kg of the proposed enzymatic composition in which sterile ultrafiltrates of Aspergilus awamori and Trichoderma viride culture fluid were taken in a ratio of 20:15. Sterile ultrafiltrates of Aspergilus awamori and Trichoderma viride culture fluid obtained from Biosynthesis AB (Viinius, Lithuania) were used in the experiment. The kneaded dough is not matured, but immediately divided, shaped and raised for 5560 minutes. Baking results are shown in Table 1.

table. Test bakings with liquid multienzyme composition

Groups no Enzyme additive, ml / kg of flour Volume of cake, cm ³ Specific volume, cm ³ / g Cellular% 1 0 (control) 1745 2.33 65.7 2 0.2 (20AA: 20TV) ' 2505 3.50 78.5 3 0.35 (20AA: 15TV) ' 2755 3.72 77.4

Composition of the multienzyme composition in relative volume units; AA Sterile filtrate of Aspergilus awamori culture: TV - Trichoderma viride culture: sterile filtrate.

From the data in Table 1 it was found that the liquid multienzyme composition improved the properties of baked bread: volume (43-57%), comparative volume (50-60%) and porosity (about 20%). Compared to the control group, the second and third group baked goods had a better appearance, the crust color was more intense brownish, increased porosity means improved pulp structure.

Test 2. Use of a dry multienzyme composition for baking white bread

The test baked with flour of the following quality: humidity - 14.8%, falling number - 314 s, gluten content - 28%, gluten index - 89 units, water (drink - 62%, dough forming time - 2 min, dough stability) - 6 min, dough dilution degree 60 FV, farinograph quality number 88 mm The same dough preparation technology was used as in the previous experiments. Baking was done in four groups. consisting of 8 relative units of xylanase, 5 relative units of glucoamylase and 1 relative unit of amylase, with the addition of 0.146 g / kg of flour. g / kg flour The multienzyme composition for this test was prepared using deep-cultured cultured liquid ultrafiltrates of Aspergilus awamori and Trichoderma reesei micromycetes produced by AB Biosynthesis ”(Vilnius, Lithuania). Fourth group with 0.125 g / kg flour baking enzyme additive Fungamyl Super AX (Novo Nordisk AS, Denmark). This enzyme supplement consists of xylanase and α-amylase enzymes mixed in certain proportions.

Baking results are shown in Table 2.

table. Test baking with dry multienzyme composition

Groups no Enzyme additive, g / kg of flour Volume of cake, cm ³ Specific volume, cm ³ / g Porosity,% 1 0 (control) 2328 ± 24 3.27 ± 0.08 78.7 ± 0.4 2 0.146 (8K: 5G: 1A) ' 2515 ± 108 3.48 ± 0.12 77.9 ± 1.3 3 0.097 (22K: 5G: 1A) ' 2503 ± 123 3.51 ± 0.14 78.5 ± 0.3 4 0.125 2370 ± 130 3.32x0.14 77.7 ± 0.4 - multienzyme compositions composition in relative a <units of volume; K -

xylanase, G - glucoamylase, A - amyiasis.

- Fungamyl Super AX (Novo Nordisk AS), an enzyme additive used for baking,

Denmark).

Pastry with a dry multienzyme composition exhibited better quality than control bakery products, with a volume greater than about 10% and a comparative volume of about 7% relative to the control. Because of the higher quality of flour used for baking, the increase in bakery quality is not as significant as with the liquid multienzyme composition (Table 1).

Experiments were carried out with the same flour to improve the properties of baked goods using a dry multienzyme composition obtained by mixing Aspergillus oryzae and Trichoderma viride dried culture liquid ultrafiltrates prepared by AB Biosynthesis. The following results were obtained: loaf volume - 2565 cm ³ , relative volume (V / G) - 3.61, porosity - 80.74%.

From the data of this test, it was found that the proposed dry multienzyme formulation exhibits the same (porosity) or better properties (bakery volume and reference volume) as the Fungamyl Super AX analogue (Novo Nordisk AS, Denmark) used for comparison.

Test 3. Use of a multienzyme composition for baking bread in combination with other bread additives

Flour with a gluten content of 28% was used in the tests, with a fall number of 37 S.7, '.'CC rKnc, .jo, 3. ,,, u, etc. iiz. π 1111 ic m j 11 {juz κ. i1 h s i h! f 1 t> H f 1 c) v f) 1 h 1 s

prepared using culture medium ultrafiltrates of deep-cultured micromycetes Aspergilus awamori and Trichoderma reesei prepared by AB Biosynthesis (Vilnius, Lithuania). The baking was performed in the first and second groups with different doses of multienzyme formulations and ascorbic acid, in the third group with only the multienzyme formulation, in the fourth group the enzyme additive Fungamyl Super AX (Novo Nordisk AS, Denmark) was used. Baking results are shown in Table 3.

table. Test bakings with dry proposed multienzyme composition and ascorbic acid

Groups no Enzymatic additive, g / kg flour H / d Specific volume, cm ³ / g Porosity,% 1 0.097 + 0.05AR ' 0.59 3.9 75.9 2 0.1455 + 0.05AR 0.61 3.9 75.3 3 0.097 0.56 3.4 72.4 4 0.125 FS 0.54 3.6 75.3

-AR = ascorbic acid;

- FS = Fungamyl Super AX.

Research results indicate that the multienzyme composition can be used with other baking additives, such as ascorbic acid. The quality of the pastries was better in this case.

Test 4. Effect of Multienzyme Composition on Bread Aging Process

The experimental bakes were made with a liquid multienzyme composition (0.2 and 0.6 ml / kg flour). The multienzyme composition for this test was prepared using culture cultured ultrafiltrates of deep-cultured micromycetes Aspergilus awamori and Trichoderma viride made by AB Biosynthesis (Vilnius, Lithuania). IBM's baking additive consists of components, malt, soybean meal, sugar, E 472e emulsifier, dextrose, ascorbic acid, and α-amylase, mixed in specific portions. For the study of the aging process, the bars were cut into slices 8 to 10 mm thick. The samples were examined on an acoustic spectrometer every hour for 6 hours after baking and for 20 hours after baking. The results of the studies are shown in Table 4.

table. Change in amplitude Ap values (kHz) of amplitude of acoustic signal passing through sample as bars age

A variation of a pastry Ap values (kHz) after baking Ratio Api / Ap ₂ o 1 or. 3 or. 6 or. 20 or. Control (baking attachment IBM, Backaldrin, Austria) 0.0759 0.0834 0.0886 0.1046 1.38 0.2 ml / kg multienzymes compositions 0.0835 0.0895 0.1005 0.1124 1.35 0.6 ml / kg multienzymes compositions 0.0751 0.0827 0.0852 0.0893 1.19

The aging rate of bakery products is not determined by the absolute values of Ap but by the dynamics of these values. The slowest increase in Aβ values for bakery products was in bars baked with 0.6 ml of multienzyme composition per kg of flour. Breads baked with 0.2 ml / kg of multienzyme composition aged at the same rate as control buns with a special anti-aging additive from IBM (Backaldrin, Austria).

Test 5. Storage stability of a liquid multienzyme composition

The assay used was a liquid multienzyme composition prepared from AB Biosynthesis AB (Vilnius, Lithuania) made by Asnerciilus awamnri and Trichoderrma viride ultrafiltrate cultures. The stability of the liquid multienzyme composition was determined by storage in a refrigerator, room temperature (18 ° C) and thermostat (37 ° C). The pH of the solution was 4.05. The results of the studies are shown in Table 5.

table. Storage stability of the liquid multienzyme composition

Activities Residual enzymatic activity (%), when stored in the refrigerator Residual enzymatic. activity (%), when stored at 18 ° C Residual enzyme activity (%) when stored at 37 ⁰ 2d 15th 2d 15th 2d 15d Xylanase 104 88 101 101 83 21st Glucoamylase 93 100 95 95 95 84 Amylase 98 93 94 97 95 81

Studies have shown that a liquid multienzyme composition retains activity for at least 15 days when stored in a refrigerator or at room temperature. When stored in a liquid multienzyme composition in a thermostat at 37 ° C, it lost most of the xylanase activity after 15 days. Thus, the proposed liquid multienzyme composition should be stored in a refrigerator or at room temperature.

The proposed multienzyme composition for improving the quality of baked goods consists of natural components - enzymes of microbiological (fungal) origin (a-amylase, pentozonase, glucoamylase and other escorting enzymes). Enzymes are a natural component of the human body: they either exist naturally in the human body or are derived from food. Enzymes of fungal origin are easy to produce in large quantities, and they are less contaminated with foreign biogenic substances compared to enzymes of animal origin. Fungal enzymes are safer because microscopic fungi do not produce endotoxins and mycotoxin producing strains are well known and are not used for industrial enzyme synthesis.

Claims (10)

DEFINITION OF INVENTION A multienzyme composition having enzymatic activities, characterized in that the activities are α-amylase, glucoamylase and xylanase in units of g / g: α-amylase 1-1000, glucoamylase 1-10000, xylanase 1-10000. Process for the preparation of a multienzyme composition according to claim 1, characterized in that it is obtained by mixing liquid or dried filtrates obtained by deep culturing of different micromycetes. Process for the preparation of a multienzyme composition according to claim 2, characterized in that the microorganism strains belong to the genera Trichoderma and Aspergilus. Process for the preparation of a multienzyme composition according to claim 3, characterized in that the micromycete of the genus Trichoderma is Trichoderma viridae or Trichoderma reesei or Trichoderma longibranchiatum. Process for the preparation of a multienzyme composition according to claim 3, characterized in that the micromycete of the genus Aspergilus is Aspergilus awamori or Aspergilus niger or Aspergilus orryzae. 6. The multienzyme composition according to claim 1, characterized in that the ratio of α-amylase to glucoamylase is preferably 1: 1 to 1:10 when baking bread without maturation. A multienzyme composition according to claim 1 for increasing the volume of bread pastries by 0.1-0.5% by weight of the dough. A multienzyme composition according to claim 1 for protecting bakery wares from 0.1 to 0.5% by weight of the dough. Multienzyme composition according to claim 1 for improving the texture of bread pastries by adding 0.1-0.5% by weight of the dough. A multienzyme composition according to claim 1 for improving the appearance of bread crusts by adding 0.1-0.5% by weight of the dough.