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Title:
IMPROVING THE ROLLABILITY OF FLAT BREADS
Document Type and Number:
WIPO Patent Application WO/2018/010966
Kind Code:
A1
Abstract:
The present invention deals with a method for improving the rollability of flat breads comprising a) adding a maltogenic alpha-amylase and a beta amylase to a flour or directly to a dough comprising a flour; b) making the dough; and c) making flat breads from the dough.

Inventors:
AUSTIN, Dilek (77 Perry Chapel Church Road, Franklinton, North Carolina, 27525, US)
Application Number:
EP2017/066146
Publication Date:
January 18, 2018
Filing Date:
June 29, 2017
Export Citation:
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Assignee:
NOVOZYMES A/S (Krogshoejvej 36, 2880 Bagsvaerd, 2880, DK)
International Classes:
A21D8/04; A21D10/00; A21D13/047; A21D13/42
Domestic Patent References:
WO1991004669A11991-04-18
WO2010124206A12010-10-28
WO1999043794A11999-09-02
WO2006032281A22006-03-30
WO2008148845A22008-12-11
WO1984002921A21984-08-02
WO1998026057A11998-06-18
WO2000032758A12000-06-08
Foreign References:
EP1541028A12005-06-15
GB2417184A2006-02-22
US20160113295A12016-04-28
GB2417184A2006-02-22
US4106991A1978-08-15
US4661452A1987-04-28
Other References:
TODD FORMAN: "Enzyme Functionality and Potential Applications for the Tortilla Industry", 20 June 2011 (2011-06-20), XP055410311, Retrieved from the Internet [retrieved on 20170926]
NEEDLEMAN; WUNSCH, J. MOL. BIOL., vol. 48, 1970, pages 443 - 453
RICE ET AL.: "EMBOSS: The European Molecular Biology Open Software Suite", TRENDS GENET., vol. 16, 2000, pages 276 - 277
H. NEURATH; R.L. HILL: "The Proteins", 1979, ACADEMIC PRESS
BOEL ET AL., EMBO J., vol. 3, no. 5, 1984, pages 1097 - 1102
AGRIC. BIOL. CHEM., vol. 55, no. 4, 1991, pages 941 - 949
GOMEZ-MENDEZ ET AL.: "Instrumental and sensory techniques for the measurement of wheat tortilla texture", IFT CONFERENCE PAPER, NEW ORLEANS, 1996
MAO: "Texture measurements of commercially available wheat flour tortillas", POSTER PRESENTED AT IFT ANNUAL MEETING, DALLAS, USA, pages 20000610
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Claims:
CLAIMS

1 . A method for improving the reliability of flat breads comprising

a) adding a maltogenic alpha-amylase and a beta amylase to a flour or to a dough comprising a flour;

b) making the dough; and

c) making flat breads from the dough. 2. The method according to claim 1 , wherein the flat breads are baked.

3. The method according to any of the preceding claims, wherein the flour is selected from the group consisting of wheat flour, corn flour, rye flour, barley flour, oat flour, rice flour, sorghum flour, soy flour, flour from pulses like gram flour, and a combination thereof.

4. The method according to any of the preceding claims, wherein the flour is corn flour.

5. The method according to any of the preceding claims, wherein the maltogenic alpha- amylase has at least 70% identity with SEQ ID NO:1.

6. The method according to any of the preceding claims, wherein the beta amylase has at least 70% identity with SEQ ID NO:2.

7. The method according to any of the preceding claims, wherein the flat bread has a reliability at 7 days post baking which is better than the reliability of a flat bread which is prepared under the same conditions, but without treatment with a maltogenic alpha-amylase and a beta- amylase.

8. The method according to any of the preceding claims, wherein additionally the flat bread has a moistness at 7 days post baking which is better than the moistness of a flat bread which is prepared under the same conditions, but without treatment with a maltogenic amylase and a beta- amylase.

9. The method according to any of the preceding claims, wherein additionally the flat bread has a softness at 7 days post baking which is better than the softness of a flat bread which is prepared under the same conditions, but without treatment with a maltogenic alpha-amylase and a beta-amylase.

10. The method according to any of the preceding claims, wherein the dough further comprises one or more enzymes selected from the group consisting of amylase, glucanase, galactanase, mannanase, aminopeptidase, alpha-amylase, carboxypeptidase, catalase, chitinase, cutinase, cyclodextrin glycosyltransferase, deoxyribonuclease, esterase, alpha- galactosidase, beta-galactosidase, glucoamylase, alpha-glucosidase, beta-glucosidase, haloperoxidase, invertase, laccase, lipase, phospholipase, mannosidase, oxidase, pectinolytic enzymes, peptidoglutaminase, peroxidase, phytase, glucose oxidase, polyphenoloxidase, proteolytic enzyme, ribonuclease, transglutaminase and xylanase.

1 1 . The method according to any of the preceding claims, wherein the flat breads are tortillas.

12. A flat bread obtainable by a method of any of the preceding claims.

13. A flat bread dough composition comprising a maltogenic alpha-amylase and a beta- amylase.

14. A flat bread dough premix comprising a maltogenic alpha-amylase, a beta-amylase, and flour selected from the group consisting of wheat flour, corn flour, rye flour, barley flour, oat flour, rice flour, sorghum flour, soy flour, flour from pulses like gram flour, and combinations thereof.

15. The flat bread dough premix according to claim 14 comprising a maltogenic alpha- amylase, a beta-amylase, and corn flour.

Description:
IMPROVING THE ROLLABILITY OF FLAT BREADS

Reference to a Sequence Listing

This application contains a Sequence Listing in computer readable form, which is incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to a process for retarding the staling of flat breads, as well as flat breads obtainable by the method of the invention. BACKGROUND OF THE INVENTION

In the bread-making process, it is known to add bread-improving and/or dough-improving additives to the bread dough in order to improve texture, volume, flavor, freshness of the bread, as well as improving machinability of the dough.

In recent years, a number of enzymes have been used as dough and/or bread improving agents.

Various amylases have been suggested for retarding the staling by the addition to dough. For example, WO 91/04669 discloses the use of a maltogenic alpha-amylase from B. stearothermophilus to retard staling.

GB 2417184 describes a process for preparation of wheat tortilla comprising using an exoamylase and an emulsifier.

WO 2010/124206 describes the use of raw starch degrading enzymes to retard staling of flat breads.

However, baked products such as flat bread provide additional challenges in retarding staling. For example, many amylases require longer baking times to allow sufficient starch modification and are therefore less suitable to the rapid baking time of flat bread.

There is therefore still a need for finding improved enzyme solutions in flat bread production.

SUMMARY OF THE INVENTION

The present invention relates to a method for improving the reliability of flat breads comprising a) adding a maltogenic alpha-amylase and a beta amylase to a flour or to a dough comprising a flour;

b) making the dough; and

c) making flat breads from the dough. In one embodiment, the flat breads are baked; especially the flat breads are baked at a different location than the dough making location.

In one embodiment, the flour according to the present invention is selected from the group consisting of wheat flour, corn flour, rye flour, barley flour, oat flour, rice flour, sorghum flour, soy flour, flour from pulses like gram flour, and a combination thereof.

In a preferred embodiment, the flour according to the present invention is corn flour.

In one embodiment, the maltogenic alpha-amylase according to the invention has at least 70% identity with SEQ ID NO:1.

In one embodiment, the beta amylase according to the invention has at least 70% identity with SEQ ID NO:2.

In one embodiment, the flat bread according to the invention has a rollability at 7 days post baking which is better than the rollability of a flat bread which is prepared under the same conditions, but without treatment with a maltogenic alpha-amylase and a beta-amylase.

In one embodiment, the flat bread according to the invention has a rollability at 14 days post baking which is better than the rollability of a flat bread which is prepared under the same conditions, but without treatment with a maltogenic alpha-amylase and a beta-amylase.

In one embodiment, the flat bread according to the invention has a moistness at 7 days post baking which is better than the moistness of a flat bread which is prepared under the same conditions, but without treatment with a maltogenic alpha-amylase and a beta-amylase.

In one embodiment, the flat bread according to the invention has a moistness at 14 days post baking which is better than the moistness of a flat bread which is prepared under the same conditions, but without treatment with a maltogenic alpha-amylase and a beta-amylase.

In one embodiment, the flat bread according to the invention has a softness at 7 days post baking which is better than the softness of a flat bread which is prepared under the same conditions, but without treatment with a maltogenic alpha-amylase and a beta-amylase.

In one embodiment, the flat bread according to the invention has a softness at 14 days post baking which is better than the softness of a flat bread which is prepared under the same conditions, but without treatment with a maltogenic alpha-amylase and a beta-amylase.

In one embodiment, the dough according to the invention further comprises one or more enzymes selected from the group consisting of amylase, glucanase, galactanase, mannanase, aminopeptidase, alpha-amylase, carboxypeptidase, catalase, chitinase, cutinase, cyclodextrin glycosyltransferase, deoxyribonuclease, esterase, alpha-galactosidase, beta-galactosidase, glucoamylase, alpha-glucosidase, beta-glucosidase, haloperoxidase, invertase, laccase, lipase, phospholipase, mannosidase, oxidase, pectinolytic enzymes, peptidoglutaminase, peroxidase, phytase, glucose oxidase, polyphenoloxidase, proteolytic enzyme, ribonuclease, transglutaminase and xylanase. In one embodiment, the flat breads according to the invention are tortillas; especially corn tortillas.

In one embodiment, we claim a flat bread obtainable by a method of the present invention.

In one embodiment, we claim a flat bread dough composition comprising a maltogenic alpha-amylase and a beta-amylase.

In one embodiment, we claim a flat bread dough premix comprising a maltogenic alpha- amylase, a beta-amylase, and a flour selected from the group consisting of wheat flour, corn flour, rye flour, barley flour, oat flour, rice flour, sorghum flour, soy flour, flour from pulses like gram flour, and a combination thereof.

In one embodiment, we claim a flat bread dough premix comprising a maltogenic alpha- amylase, a beta-amylase, and corn flour.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

Sequence identity: The relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter "sequence identity".

For purposes of the present invention, the sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et ai, 2000, Trends Genet. 16: 276-277), preferably version 5.0.0 or later. The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of Needle labeled "longest identity" (obtained using the -nobrief option) is used as the percent identity and is calculated as follows:

(Identical Residues x 100)/(Length of Alignment - Total Number of Gaps in Alignment) Variant: The term "variant" means a polypeptide having either maltogenic alpha-amylase activity or beta-amylase activity comprising an alteration, i.e., a substitution, insertion, and/or deletion, at one or more (e.g., several) positions. A substitution means replacement of the amino acid occupying a position with a different amino acid; a deletion means removal of the amino acid occupying a position; and an insertion means adding one or more amino acids adjacent to and immediately following the amino acid occupying a position.

Improved property: When the enzyme composition according to the invention is incorporated into a flour and/or a dough in effective amounts, one or more properties are improved compared to a flour and/or a dough in which the enzyme composition is not added. The improved property may be determined by comparison of a dough and/or a baked product prepared with and without addition of the enzyme composition of the present invention in accordance with the methods of present invention which are described below. Organoleptic qualities may be evaluated using procedures well established in the baking industry, and may include, for example, the use of a panel of trained taste-testers.

Flat Bread and Flat Bread Dough Compositions

As used herein, "flat bread" means bread prepared from flattened dough, and which typically has a thickness of one millimeter to a few centimeters. A flat bread may be made from a simple mixture of flour, water, and salt and then thoroughly rolled into flattened dough. Flat breads have a very quick baking time (often <2 minutes).

In one embodiment, the flat bread is unleavened, i.e., made without yeast.

In another embodiment, the flat bread is made with yeast. The flat bread may include further optional ingredients, such as olive oil, sesame oil, shortenings, spices, garlic, curry powder, diced jalapefios, chili powder, pepper, vegetables, and the like.

Examples of flat breads include tortilla, pita, Arabic bread, Indian flat bread (IFB), wheat and gluten free flat breads. Further non-limiting examples include lavash, baladi, barbari, Sangak, tanoor, taftoon, shami, halabi, mafrood, burr, bairuti, pocket bread, naan, phulka, chapatti, and paratha.

In a particular embodiment, the flat bread product is a tortilla, especially corn tortilla.

As used herein "flat bread dough" means any dough used to prepare a flat bread.

The dough used to prepare a flat bread product may be made from any suitable flour source, e.g., flour sourced from grains, such as, wheat flour, corn flour, rye flour, barley flour, oat flour, rice flour, or sorghum flour, potato flour, soy flour, flour from pulses and combinations thereof; in particular corn flour.

According to the present invention, the corn flour may be produced as known in the art, e.g., to produce nixtamalized corn flour (NCF). Nixtamalized corn flour is produced by the steps of alkaline cooking (heating and steeping) of corn, washing, wet milling the nixtamal, and drying, thereby producing corn masa flour. Any flat bread process may be used to prepare the flat bread. The process of preparing flat bread generally involves the sequential steps of dough making (with an optional proofing step), sheeting or dividing, shaping or rolling, and proofing the dough, which steps are well known in the art.

In addition to preparing fresh flat bread dough or flat bread products, the present invention is also directed to a method for preparing flat bread dough that can be stored, e.g., at room temperature or with refrigeration, prior to baking. An example of a method for preparing a flat bread dough that can be stored prior to baking includes the steps of making a dough (with an optional proofing), sheeting or dividing, shaping or rolling, proofing, and storing the dough.

In addition to preparing fresh flat bread dough or flat bread products, the present invention is directed to a method for preparing a frozen flat bread dough. The dough is frozen after preparation of the dough comprising the enzyme combination of the present invention (i.e., prior to baking). A frozen flat bread dough may be advantageous for storage and/or distribution. An example of a method for preparing a frozen flat bread dough includes the steps of making a dough (with an optional proofing), sheeting or dividing, shaping or rolling, proofing, and freezing the dough. The present invention is also directed to a frozen flat bread dough comprising the enzyme combination of the present invention.

In one embodiment, the invention provides a method for preparing a flat bread composition, comprising a maltogenic alpha-amylase and a beta amylase.

Industrial Processes

The present invention is particularly useful for preparing flat bread dough and flat bread products in industrialized processes, that is, in which the dough used to prepare flat bread and/or flat bread products are prepared mechanically using automated or semi-automated equipment.

The present invention provides significant advantages in that flat bread can now be prepared using automated or semi-automated processes in which the flat bread is stored for distribution and consumer use more than 24 hours after preparation.

The process of preparing flat bread generally involves the sequential steps of dough making (with optional proofing step(s)), sheeting or dividing, shaping or rolling, and proofing, the dough, which steps are well known in the art. In an industrial flat bread production process according to the present invention, one or more of these steps is/are performed using automated or semi-automated equipment.

The present invention is particularly useful for corn tortilla processing, in particular cold press. Enzymes

Maltogenic alpha-amylase

The maltogenic alpha-amylase according to the invention (EC 3.2.1.133) may be from Bacillus. A maltogenic alpha-amylase from B. stearothermophilus strain NCIB 1 1837 is commercially available from Novozymes A S under the tradename NOVAMYL.

Preferably, the maltogenic alpha-amylase is an enzyme having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the polypeptide of SEQ ID NO: 1. Preferably, the maltogenic alpha-amylase is an enzyme having the amino acid sequence shown in SEQ ID NO:1 herein.

The maltogenic alpha-amylase may further comprise one or more additional alterations at one or more (e.g., several) other positions in SEQ ID:1.

The amino acid changes may be of a minor nature, that is conservative amino acid substitutions or insertions that do not significantly affect the folding and/or activity of the protein; small deletions, typically of 1 -30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue; a small linker peptide of up to 20-25 residues; or a small extension that facilitates purification by changing net charge or another function, such as a poly- histidine tract, an antigenic epitope or a binding domain.

Examples of conservative substitutions are within the groups of basic amino acids (arginine, lysine and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine and valine), aromatic amino acids (phenylalanine, tryptophan and tyrosine), and small amino acids (glycine, alanine, serine, threonine and methionine). Amino acid substitutions that do not generally alter specific activity are known in the art and are described, for example, by H. Neurath and R.L. Hill, 1979, In, The Proteins, Academic Press, New York. Common substitutions are Ala/Ser, Val/lle, Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr, Ser/Asn, Ala/Val, Ser/Gly, Tyr/Phe, Ala/Pro, Lys/Arg, Asp/Asn, Leu/lle, LeuA al, Ala/Glu, and Asp/Gly.

The maltogenic alpha-amylase may also be a variant of the maltogenic alpha-amylase from B. stearothermophilus as disclosed in, e.g., W01999/043794; WO2006/032281 ; or WO2008/148845, e.g., Novamyl® 3D available from Novozymes A/S.

A maltogenic alpha-amylase is added in an effective amount for retarding the staling of the baked product. The amount will typically be in the range of 0.01 -10 mg of enzyme protein per kg of flour, e.g., 0.05-10 mg of enzyme protein per kg of flour. Beta amylase

The beta-amylase according to the present invention (EC 3.2.1.2) is preferably selected from the group consisting of a Bacillus flexus beta-amylase, a Clostridium thermosulphurogenes beta-amylase, a barley beta-amylase, or a wheat beta-amylase. In a preferred embodiment, the beta-amylase is a Bacillus flexus beta-amylase.

More particularly, the beta-amylase comprises the polypeptide of SEQ ID NO: 2; or a beta-amylase having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the polypeptide of SEQ ID NO: 2.

The beta amylase may further comprise one or more additional alterations at one or more

(e.g., several) other positions in SEQ ID:2.

The amino acid changes may be of a minor nature, that is conservative amino acid substitutions or insertions that do not significantly affect the folding and/or activity of the protein; small deletions, typically of 1 -30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue; a small linker peptide of up to 20-25 residues; or a small extension that facilitates purification by changing net charge or another function, such as a poly- histidine tract, an antigenic epitope or a binding domain.

Examples of conservative substitutions are within the groups of basic amino acids (arginine, lysine and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine and valine), aromatic amino acids (phenylalanine, tryptophan and tyrosine), and small amino acids (glycine, alanine, serine, threonine and methionine). Amino acid substitutions that do not generally alter specific activity are known in the art and are described, for example, by H. Neurath and R.L. Hill, 1979, In, The Proteins, Academic Press, New York. Common substitutions are Ala/Ser, Val/lle, Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr, Ser/Asn, Ala/Val, Ser/Gly, Tyr/Phe, Ala/Pro, Lys/Arg, Asp/Asn, Leu/lle, LeuA al, Ala/Glu, and Asp/Gly.

The beta-amylase may be the Secura™ product obtainable from Novozymes A S.

A beta-amylase is added in an effective amount for retarding the staling of the baked product. The amount will typically be in the range of 0.01 -10 mg of enzyme protein per kg of flour, e.g., 0.05-10 mg of enzyme protein per kg of flour.

Additonal enzymes

Optionally, one or more additional enzymes, such as amylase, glucanase, galactanase, mannanase, aminopeptidase, alpha-amylase, carboxypeptidase, catalase, chitinase, cutinase, cyclodextrin glycosyltransferase, deoxyribonuclease, esterase, alpha-galactosidase, beta- galactosidase, glucoamylase, alpha-glucosidase, beta-glucosidase, haloperoxidase, invertase, laccase, lipase, phospholipase, mannosidase, oxidase, pectinolytic enzymes, peptidoglutaminase, peroxidase, phytase, glucose oxidase, polyphenoloxidase, proteolytic enzyme, ribonuclease, transglutaminase or xylanase may be used together with the enzyme composition according to the invention. The additional enzyme may be of any origin, including mammalian and plant, and preferably of microbial (bacterial, yeast or fungal) origin.

The glucoamylase for use in the present invention include glucoamylases having a sequence identity of at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% to the amino acid sequence of the A. niger G1 or G2 glucoamylase (Boel et al. (1984), EMBO J. 3 (5), p. 1097-1 102), the A. awamori glucoamylase disclosed in WO 84/02921 , or the A. oryzae glucoamylase (Agric. Biol. Chem. (1991 ), 55 (4), p. 941 -949). A suitable commercial glucoamylase is GoldCrust™ obtainable from Novozymes A/S.

Suitable commercial alpha-amylase compositions include, e.g., BAKEZYME P 300 (available from DSM) and FUNGAMYL 2500 SG, FUNGAMYL 4000 BG, FUNGAMYL 800 L, FUNGAMYL ULTRA BG and FUNGAMYL ULTRA SG (available from Novozymes A/S).

The glucose oxidase may be a fungal glucose oxidase, in particular an Aspergillus niger glucose oxidase (such as GLUZYME®, available from Novozymes A/S).

The xylanase which may be of microbial origin, e.g., derived from a bacterium orfungus, such as a strain of Aspergillus, in particular of A. aculeatus, A. niger, A. awamori, or A. tubigensis, from a strain of Trichoderma, e.g. T. reesei, or from a strain of Humicola, e.g., H. insolens.

Suitable commercially available xylanase preparations for use in the present invention include PANZEA BG, PENTOPAN MONO BG and PENTOPAN 500 BG (available from Novozymes A/S), GRINDAMYL POWERBAKE (available from Danisco), and BAKEZYME BXP 5000 and BAKEZYME BXP 5001 (available from DSM).

The protease may be from Bacillus, e.g., B. amyloliquefaciens.

The phospholipase may have phospholipase A1 , A2, B, C, D or lysophospholipase activity; it may or may not have lipase activity. It may be of animal origin, e.g., from pancreas, snake venom or bee venom, or it may be of microbial origin, e.g., from filamentous fungi, yeast or bacteria, such as Aspergillus or Fusarium, e.g., A. niger, A. oryzae or F. oxysporum. A preferred lipase/phospholipase from Fusarium oxysporum is disclosed in WO 98/26057. Also, the variants described in WO 00/32758 may be used.

Suitable phospholipase compositions are LIPOPAN F and LIPOPAN XTRA (available from Novozymes A/S) or PANAMORE GOLDEN and PANAMORE SPRING (available from DSM).

Enzyme Treatment The enzymes are added to the flat bread dough ingredients (i.e., prior to baking or freezing the dough), e.g., indirectly to the dough by adding it to the flour used to prepare the dough, or directly to the dough itself.

The enzymes may be added to flour or dough in any suitable form, such as, e.g., in the form of a liquid, in particular a stabilized liquid, or it may be added to flour or dough as a substantially dry powder or granulate. Granulates may be produced, e.g., as disclosed in US Patent No. 4,106,991 and US Patent No. 4,661 ,452. Liquid enzyme preparations may, for instance, be stabilized by adding a sugar or sugar alcohol or lactic acid according to established procedures. Other enzyme stabilizers are well-known in the art. The enzyme combination treatment may be added to the flat bread dough ingredients in any suitable manner, such as individual components (separate or sequential addition of the enzymes), or addition of the enzymes together in one step or one composition.

The dough may also comprise other conventional ingredients, e.g., one or more emulsifiers. Emulsifiers serve to improve dough extensibility and may also be of some value for the consistency of the resulting flat bread, as well as for its storage stability and handling. Examples of suitable emulsifiers are mono- or diglycerides, polyoxyethylene stearates, diacetyl tartaric acid esters of monoglycerides, sugar esters of fatty acids, propylene glycol esters of fatty acids, polyglycerol esters of fatty acids, lactic acid esters of monoglycerides, acetic acid esters of monoglycerides, lecithin or phospholipids, or ethoxylated monoglycerides. Particular emulsifiers include monoglycerides, diacetyl tartaric acid esters of monoglyceride (DATEM) and sodium stearoyl lactylate (SSL).

Other conventional ingredients include proteins, such as milk powder, gluten, and soy; eggs (either whole eggs, egg yolks or egg whites); an oxidant such as ascorbic acid, potassium bromate, potassium iodate, azodicarbonamide (ADA), ammonium persulfate or potassium persulphate; an amino acid such as L-cysteine; a sugar such as sucrose, dextrose, etc.; a salt such as sodium chloride, calcium acetate, sodium sulfate or calcium sulfate, diluents such silica dioxide, starch of different origins. Still other convention ingredients include hydrocolloids such as CMC, guar gum, xanthan gum, locust bean gum, etc. Modified starches may be also used.

Pre-Mixes

It will often be advantageous to provide the enzymes used in the treatment of the present invention in admixture with other ingredients used to improve the properties of flat bread products. These are commonly known in the art as "pre-mixes," which usually comprise flour.

Hence, in a further aspect, the present invention relates to a flat bread premix for improving the quality of dough used to prepare a flat bread product or flat bread products, which premix comprises a maltogenic alpha-amylase and a beta amylase and one or more flat bread dough ingredients, in particular flour such as flour from grains, such as, wheat flour, corn flour, rye flour, barley flour, oat flour, rice flour, sorghum flour, soy flour, or flour from pulses like gram flour, and combinations thereof. In a preferred embodiment, the flour is corn flour. In another embodiment, the present invention relates to a flat bread pre-mix comprising the enzyme combinations of the present invention and flour, such as, flour from grains, such as, wheat flour, corn flour, rye flour, barley flour, oat flour, rice flour, sorghum flour, and combinations thereof, and one or more additional enzymes, as previously described.

The pre-mix composition may be in liquid form or dry or substantially dry form.

Dough and Flat Bread Properties

In one embodiment, the flat bread prepared by the methods and compositions of the invention provides improved storage properties. The flat bread prepared by the methods and compositions of the present invention are used as anti-staling agents to improve the shelf life of the flat bread product. The anti-staling effect (and improved shelf life) of a flat bread product can be determined by a number of methods well known in the art.

Primarily anti-staling effectiveness is measured by the hardness (also referred to as "firmness" and the opposite of "softness") of the flat bread product. Hardness can be measured using a texture profile analyzer. Texture measurements for flat bread such as tortilla can be measured according to methods known in the art as disclosed in e.g., Gomez-Mendez et al. "Instrumental and sensory techniques for the measurement of wheat tortilla texture." IFT Conference Paper, New Orleans (1996) and Mao, "Texture measurements of commercially available wheat flour tortillas." Poster presented at IFT Annual Meeting, Dallas, USA (June 10- 14, 2000). Besides hardness/softness, stickiness, extensibility and elasticity are also important quality parameters for flat bread. Other important properties include reliability, foldability, flexibility, layering, bite and/or texture.

Other tests known in the art may be used to assess the shelf life and other organoleptic qualities of the flat bread prepared by the methods and compositions of the present invention. Storage/Shelf Life

In one embodiment, the present invention relates to a flat bread having an improved shelf life at least 1 hour after baking. In one embodiment, the present invention relates to a flat bread having an improved shelf life at least 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 1 1 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours or 23 hours after baking. In one embodiment, the present invention relates to a flat bread having an improved shelf life at least 24 hours after baking. In another embodiment, the present invention relates to a flat bread having an improved shelf life at least 48 hours after baking. In another embodiment, the present invention relates to a flat bread having an improved shelf life at least 72 hours after baking. In another embodiment, the present invention relates to a flat bread having an improved shelf life at least 96 hours after baking. In another embodiment, the present invention relates to a flat bread having an improved shelf life at least 120 hours after baking. In another embodiment, the present invention relates to a flat bread having an improved shelf life at least 6 days, 7 days, 8 days, 9 days, 10 days, 1 1 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days or 21 days after baking.

Shelf life can be measured as follows: A flat bread is prepared using enzyme compositions of the present invention and compared to a control flat bread, that is, a flat bread prepared in the same way but without enzyme compositions of the present invention. The flat bread is stored in a sealed plastic bag at 25 ° C. After the storage period, (e.g., 1 hour, 24 hours, 48 hours, 72 hours, 96 hours, 7 days, 21 days etc.), the hardness of the flat bread is measured using a texture analyzer and compared to a control flat bread stored under identical conditions. An improved shelf life is defined as a flat bread which is less hard (i.e., softer) than the control as measured by the texture analyzer. In addition to preparing fresh flat bread dough or flat bread products, the present invention is directed to a method for preparing flat bread dough that can be stored, e.g., at room temperature or with refrigeration, or frozen prior to baking. The dough can be stored and/orfrozen after preparation of the dough and treatment by the enzyme combinations of the present invention (i.e., prior to baking) for 1 hour, 24 hours, 48 hours, 72 hours, 96 hours, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 1 1 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, etc.

In one embodiment, the flat bread is also compared to a control in other quality parameters, such as, stickiness, extensibility, elasticity, reliability, foldability, flexibility, layering, bite and texture; especially reliability. The flat bread prepared by the enzyme treatment of the present invention is analyzed at a time after baking or during storage (e.g., 1 hour after baking and/or 24 hours, 48 hours, 72 hours, 96 hours, 7 days, 14 days, 21 days, etc. post baking).

The flat bread prepared by the enzyme treatment of the present invention preferably has improved qualities in terms of improved stickiness, extensibility, elasticity, reliability, flexibility, foldability, layering, bite and/or texture; especially improved rollability as compared to treatment with other anti-staling enzymes.

The flat bread may be prepared with other background enzymes.

The invention described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed, since these embodiments are intended as illustrations of several aspects of the invention. Any equivalent embodiments are intended to be within the scope of this invention as well as combinations of one or more of the embodiments. Various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.

Various references are cited herein, the disclosures of which are incorporated by reference in their entireties. The present invention is further described by the following examples which should not be construed as limiting the scope of the invention. For example, routine modifications to optimize the methods of enzymatic modification according to the present invention are contemplated.

EXAMPLES

Example 1

Rollability of corn tortillas Corn tortillas were made in the following way: Mixture I:

0.5 kg corn masa flour;

40 ppm Novamyl 10000 BG;

and 0.6 kg water were mixed for 5 minutes.

Mixture II:

0.5 kg corn masa flour;

460 ppm Secura;

and 0.6 kg water were mixed for 5 minutes.

Mixture III:

0.5 kg corn masa flour;

40 ppm Novamyl 10000 BG and 460 ppm Secura; and 0.6 kg water were mixed for 5 minutes.

40 g dough balls were made from the above mixture(s).

The dough balls rested for 5 minutes at room temperature (covered with plastic).

The dough balls were shaped into thin circular disk using a cold pressing techniqu

The thin disk of dough had a diameter of 18 cm.

The thin disk of dough was baked for 1 min. into tortillas (30 sec. for each side). The tortillas were cooled for 15 min.

The tortillas were evaluated at day 1 , day 7, and day 14 (after baking).

10 trained people evaluated the tortillas.

Results/Table 1 :

Rollability scores at day 1 ; day 7; and day 14 Scoring 1 -5:

1 very easy to break when it is rolled; and

Enzyme 5 being very flexible to roll without any cracks

Day 1

maltogenic alpha-amylase 3.88

beta amylase 4.00

maltogenic alpha-amylase+ beta

amylase 3.85

Day 7

maltogenic alpha-amylase 2.37

beta amylase 2.72

maltogenic alpha-amylase+ beta

amylase 3.24

Day 14

maltogenic alpha-amylase 1 .93

beta amylase 2.38

maltogenic alpha-amylase+ beta

amylase 2.78 Conclusion:

The rollability of the tortillas made with the mixture of maltogenic alpha-amylase and beta-amylase is surprisingly good after 7 days and 14 days, compared with the rollability of the tortillas made with maltogenic alpha-amylase alone, or beta-amylase alone.