MXPA97001998A - Pasta or mass cookies, cookie products and methods to produce my - Google Patents

Abstract

The present invention describes an enzymatic composition comprising, a protease which is inactivated by oxidation by an oxidizing agent, and an enzyme which produces the oxidizing agent. This composition can be used to soften pastes or doughs for the production of biscuits or biscuits

Description

A NEW ENZYMATIC COMBINATION FIELD OF THE INVENTION The present invention relates to a composition comprising an enzyme which produces an oxidizing agent and a protease which is inactive by that agent. These compositions find use in pastes or masses for cooking.

BACKGROUND OF THE INVENTION Metabisulfite is commonly used in the cooking or baking industry to soften pastes or doughs. In particular, sulfite is used in the cookie industry to reduce the shrinkage or shrinkage of dough pieces or dough and the irregular sizing of baking or bakery products. The masses or pastes contain as a minimum flour and water although these can, of course, contain yeast, sugar, enzymes, sodium bicarbonate, etc. Sulfite is thought to react with gluten proteins in a way that prevents them from the formation of covalent SS bridges (CE Stauffer (1994).) REF: 24319 Science of Cookie and Cracker Production by Hamed Faridi, Chap an & Hall New York London, Chapter 6. pp. 237-23.8). The effect of sulfite in pasta or dough is almost immediate and results in a paste or inextensible and inelastic mass. Sulfite also activates wheat proteases which increase the breakdown or separation of the gluten structure (H.S. Olcott, L.A. Sapirstein, M.J. Blish, Cereal Chem. (1943) 20 (1), 87-97). Cysteine and glutathione were also shown to have similar effects (C.O. t Swanson, A.C. Andrews, Cereal Chem. (1945) 22 (3), 134-149). Papain was one of the first enzymes applied for the modification of wheat gluten (C.O. Swanson, A.C. Andrews, Cereal Chem. (1945) 22 (3), 134-149, R.H. Harris, J. Jr. Johnson, Cereal Chem. (1940) 17 (3), 203-222). The use of microbial proteases kt * £ r has also been described in many patents: US Patent 3,157,513, US Patent 20 1,377,798, US Patent 4,100,151, British Patent 2007960, and German Patent Application DE 3003679 A1. Microbial proteases can be combined with pig pancreas enzymes as described in EP 0384303. Enzymatic hydrolysis Partial gluten of wheat has also been described using proteases of Thermoactinomyces vulgaris as described by M. Friedrich, J. Noack, R. Noack, Die Nahrung (1982) 26 (9) 811-822; J.I. Tschimirov, K.D. Schweinke, D. Augustat, V. Tolstoguzov, Die Nahrung (1983) 27 (7) 659-668. Compared with sulfite, proteases work in a different way since they hydrolyse the gluten peptide bonds. This also decreases the degree of break or separation of pieces of pasta, .10 or mass and provides a more regular sizing * of cookies or biscuits. However, the action of such proteases is time dependent. This is the biggest limiting factor in the use of proteases in pastes or doughs because the manufacturers of biscuits or biscuits need some degree of freedom or ease with respect to the resting time of doughs or pastes. This is possible with the rapid effect of reducing agents similar to sulfites but ? * without it being easy to control with continuous action of the proteases.

DESCRIPTION OF THE INVENTION Surprisingly it has been found that a novel combination of enzymes can allow manufacturers of biscuits or biscuits to mimic the '' 'sulfite effect in paste or dough. In accordance with the present invention, a combination of a protease which is inactive by oxidation and an enzyme capable of giving rise or origin to this oxidation. This combination of enzymes can replace the metabisulfite in dough or dough, for example, dough or baking dough such as in the production of biscuits or biscuits. • The oxidation sensitive protease is preferably a thioprotease, for example papain or bromelain. The oxidizing enzyme preferably produces an oxidizing agent such as H? 0? (Hydrogen peroxide) which can inactivate the protease after a certain period of time. Preferably the enzyme is glucose oxidase, sulfhydryl oxidase or amino acid oxidase. Good results can be obtained with papain, such as from Carica papaya, commercially available from Gist 20 Brocades under the trademark Protease V100, combined with a (for example fungicide) glucose oxidase preferably from Aspergillus niger, commercially available from Gist Brocades under the trademark Maxazyme GO 1500. 25 Using a protease that can only be active at the beginning of the preparation of the dough or pasta, the shrinkage or contraction of the dough or pasta can be reduced and more regular sizes of the baking products or of bakery, such as biscuits or cookies, can be obtained. The action of (or each) protea = a can then be substantially diminished when the concentration of the oxidizing agent (s) has reached a particular level (of inactivation). Therefore, the required amount of enzyme necessary for the production of sufficient oxidizing agent, may be a function of the oxidation stability of the protease, the amount of protease present, the effectiveness of the oxidizing agent and the desired time after which the Protease activity should be decreased to a desired level. The protease activity (NF) is determined by the hydrolysis of casein at pH 6.0, at 40 ° C for 60 minutes. One unit of NF is the amount of enzyme needed to release the equivalent of 1? Q tyrosine per hour after precipitation of the remaining proteins with trichloroacetic acid. Oxidase activity can generally be determined by oxidizing a substrate in a buffer (pH of about 5.4) at a temperature of approximately 37 ° C for 10 minutes. t The hydrogen peroxide produced is measured in the presence of horseradish peroxidase and o-dianisidine dihydrochloride. 1 SU is the amount 5 of enzyme needed to consume 0.4 / junk oxygen / minute. In general, 10 5 to 109 NF / kg of flour, 6 8 preferably 10 to 10 NF / kg of protease meal are added to (or present in) the paste or mass. Of the enzyme that produces the oxidizing agent < for example glucose oxidase, 50 to 50000 SU / kg of flour, preferably 100 to 2000 SU / kg of flour are added to (or are present in) the dough or dough. Taking glucose oxidase as an example, glucose oxidase activity (GOX) is determined by oxidizing glucose (0.11 M) in 0.1 M phthalate buffer (pH 5.4) at 37 ° C for 10 minutes. £ -k ~ minutes in the presence of horseradish peroxidase (40 mg / 1 POD-II, available from Boehringer Mannheim) and o-dianisidine dihydrochloride (130 mg / 1). 1 SU is the amount of enzyme needed to consume 0.4 ipoles of oxygen / minute under the conditions of the test. Sulfhydryl oxidase (SOX) can not be determined by the above method because the hydrogen peroxide reacts with the SOX substrate (glutathione). Of course, the activity of sulfhydryl oxidase is determined by measuring the decrease of the glutathione of the substrate as described by Young and Nimmo, Biochem. J. 130 (1972) 33. A unit of sulfhydryl oxidase is equal to an amount of enzyme required to deplete or decrease 1 / mole of oxygen / minute from a test mixture containing 8 mmole of GSH. (glutathione) and 40 mmoles sodium acetate (pH. * 5.5) at 25 ° C. The amount of hydrogen peroxide produced per 1 unit of SOX is approximately the same as for 1 unit of GOX (SU). 15 Use of the Farinograph and its interpretation.

The farinograph measures and records the resistance £ 5"of a dough or pasta during mixing.

In the apparatus it is possible to measure the effect of compounds that affect the consistency of a dough or paste, such as etabisulfite and protease. A consistency of approximately 500 BU (Brabander Units) is a good consistency for baking bread. When the gluten In a paste or dough is hydrolyzed by a protease, the resulting mixture containing starch and hydrolyzed protein has a final consistency of 100 to 200 BU. In order to bake the biscuit or biscuit, the desired consistency is between the consistency of a dough or bread dough and a fully hydrolyzed dough or dough, for example preferably from 300 to 400 BU. Unit DS .. _ is the decrease in the curve of the farinograph between the maximum value and 15 minutes after the maximum value. The invention will now be described, by way of illustration only, with reference to the following Examples and drawings, in which: Figure 1 is the farinogram of the test dough or no. 1 of Example 1; Figure 2 is the farinogram of the test dough or no. 2 of Example 1; and Figure 3 is the farinogram of the test dough or no. 4 of Example 1.

Example 1 A dough or dough was prepared from wheat flour by mixing 300 g of flour and water (final volume 188 ml) for at least 20 minutes in a farinograph, as a control or with a protease and oxidant enzyme. For tests in which GOX was added, the dough or pasta was also supplemented with glucose (2 g / kg flour). Several parameters were measured for four masses or pastes, the results of which are shown in the following Table.

Table 1 DS1C .: degree of softness or softening after 15 minutes in BU (Brabender Units) BUanc, hurricane: the width of the trace of the farinog ^ bouquet after 15 minutes of mixing time, Typically the effect of the proteases is to decrease this value producing a narrow or thin line in the farinogram. The results show that glucose oxidase was able to reduce the action of papain. -r From the figures it is clear that papain r-hydrolyzes gluten too much, so that the resulting dough or dough is not suitable for baking biscuits or cookies. The combination of papain and GOX, however, results in a rapid decrease in consistency to a desirable level. The level remains more or less constant during the time. Prolonged mixing can even result in an increase in consistency, possibly due to indirect oxidation of gluten by H-0- produced by GOX.

EXAMPLE 2 Influence of glucose concentration The tests were carried out as described in Example 1, test No. 4 but with varying amounts of glucose in the pastes or masses: & w Table 2 20 Optimal levels of papain and GOX are dependent on the recipe and process conditions. For example, the glucose level influences GOX activity as can be observed from the results shown in Table 2. Glucose is already present in the flour (without any complementation). Therefore, the combination of GOX and papain can work without adding glucose, if necessary. The results show that the addition of glucose allows the pasta or dough to recover some concentration (consistency) sooner. As can be seen from the farinograph, the consistency first decreases and then increases at the end to a higher value than when GOX was only present. Probably, only part of the hydrogen peroxide produced by GOX is used to eliminate papain, part is also being used for peroxidases in the flour to strengthen the gluten network at the end. The effect of glucose is when more hydrogen peroxide is produced which causes the recovery of concentration of the mass or paste to occur faster.

Example 3 f * - Influence of GOX concentration The tests were carried out as described in Example 1 test No. 4, except that the amounts of GOX added to the paste or dough were varied (papain was present at the level in No. 4 of Example 1).

Table 3 10 fifteen The results shown in Table 3 indicate that with more GOX, the consistency of the paste or dough has a higher value.

Example 4 Stability of the pastes or masses The tests were carried out as described in Example 1 tests Nos. 2 and 4, and were compared with a sulfite control. The mixing time was 15 minutes in a first step; the viscosity of the dough or dough was measured after 1 h and 5 h of resting times. The glucose was presented in pastes or masses at 2 g / kg of flour.

Table 4 In test No. 3, 200 ppm of sulfite were used. The doses in the preparation of biscuits or biscuits can vary from 200 to 1200 ppm depending on the products and processes involved.

BU5 .., n will be lower with a higher dose of sulfite. The effect of GOX in combination with papain was to concentrate the paste or dough enough to stabilize it in a similar way to sulfite.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention is that which is clear from the present description of the invention.

Having described the invention as above, the contents of the following are claimed as property. fifteen t? - * twenty

Claims (14)

1. CLAIMS r * 1. An enzymatic composition characterized in that it comprises: (a) a protease that is at least partially inactive in oxidation by an oxidizing agent; and (b) an enzyme which produces that oxidizing agent.
2. 2. A composition according to claim 1, characterized in that the protease is a thioprotease, optionally papain or bromelain,
3. 3. A composition according to claim 1 or 2, characterized in that the oxidizing agent is H2 ° 2".
4. ?? 4. A composition according to any of the preceding claims, characterized in that the enzyme that produces the oxidizing agent is glucose oxidase, sulfhydryl oxidase or amino acid oxidase.
5. 5. A composition according to any of the preceding claims, characterized in that the protease is papain obtained from Carica papaya.
6. 6. A composition according to any of the preceding claims characterized in that the enzyme in (b) is glucose oxidase obtained from Aspergillus niger.
7. 7. A paste or dough suitable for baking or for cooking, characterized in that it comprises an enzymatic composition according to any of the preceding claims and optionally other ingredients of the dough or dough.
8. 8. A paste or dough according to claim 7, characterized in that the protease is present from 10 6 to 107 NF / kg of flour.
9. 9. A paste, or dough according to claim 7 or 8, characterized in that the enzyme is present from 500 to 1500 SU / kg of flour.
10. 10. A method of making a dough or dough suitable for baking or baking, the process is characterized in that it comprises mixing the following ingredients: (a) a protease that is at least partially inactive by an oxidizing agent; (b) an enzyme that produces that oxidizing agent; 5 (c) flour; and (d) water.
11. 11. A method according to claim 10, characterized in that it comprises adding an enzymatic composition according to any of claims 1 to 6, to a paste or dough comprising flour and water.
12. 12. A process for producing a bakery product 15, the process is characterized in that it comprises: (i) providing a dough or dough which comprises < && amp; (a) a protease that is at least partially inactive in oxidation by an oxidizing agent; (b) an enzyme which produces that oxidizing agent; (c) flour and water; and (ii) bake the dough or dough. 25
13. 13. A bakery product produced by a process according to claim 12.
14. 14. The use of an enzyme which produces an oxidizing agent that, by oxidation, inactivates a protease to make the dough or dough or in the preparation of a bakery product.