Method for preparing reduced-fat baked products

Abstract

The present invention relates to methods, uses, and products for preparing baked products containing reduced fat. The method comprises the steps of: a) preparing a dough using one or more lipases and / or one or more proteases; b) shaping and / or sheeting the dough; and c) baking the dough. The baked product is selected from the group consisting of biscuits, cookies, and crackers. The baked product has at least 10-25% reduced fat by weight when compared to a baked product prepared without the one or more lipases and / or one or more proteases.

Description

[Technical Field]

[0001] Sequence Listing Reference This application contains a Sequence Listing in computer readable form, which is incorporated herein by reference.

[0002] The present invention relates to a method for preparing baked products, and in particular to a method for preparing baked products containing reduced fat. [Background technology]

[0003] The bakery industry is one of the largest industries in the food processing sector, and the biscuit industry is the largest segment of the bakery industry due to its nutritional profile, ready-to-eat product, rich energy content, excellent source of dietary supplements, easy digestibility, and high hygienic properties. Typically, the basic ingredients of biscuits and cookies are flour, oil, and sugar. Other ingredients may vary depending on the type of biscuit or cookie and the recipe used. Most biscuits and cookies are high in fat and calories. Currently, there is considerable research into the contribution of commercially baked biscuits and cookies to the rise in obesity, especially childhood obesity. The industry is focused on providing commercially baked biscuits and cookies with lower calorie content by simply reducing the fat and / or sugar content in the biscuit or cookie formula. Reducing the fat content of the recipe may result in baked biscuits and cookies that are drier, more brittle, and have otherwise unacceptable properties and a shorter shelf life.

[0004] The fat component of any dough acts to smooth the dough, extend the shelf life of commercially baked biscuits and cookies, and improve the texture of the biscuits and cookies. The shelf life of commercially baked biscuits and cookies is extended primarily by adding fat throughout the dough to spread and coat the sugar component of the dough, thereby preventing air from entering the sugar component. However, reducing the fat content also reduces the ability of the remaining fat components to spread and coat the sugar.

[0005] In addition, the cost of various types of oils used in baking, such as soybean oil, palm oil, sunflower oil, rapeseed oil, safflower oil, corn oil or cottonseed oil, is increasing for various reasons. By reducing the amount of oil used in baking biscuits and cookies, the overall price for producing biscuits and cookies can be reduced.

[0006] Although it is known from the prior art to use phospholipids, emulsifiers or modified starches of organic origin for baking biscuits and cookies, such substitutes are used in large amounts, approximately 3% or more, which are expensive and furthermore impair the texture, mouthfeel and taste of the baked biscuits and cookies.

[0007] Therefore, there is a need to cost-effectively reduce the fat and / or sugar content in baked biscuits and cookies without compromising flavor, texture and desirable taste. Summary of the Invention [Means for solving the problem]

[0008] The present invention relates to a method for preparing a baked product.

[0009] In one aspect, the present invention relates to a method for preparing a reduced-fat baked product, the method comprising the steps of a) preparing a dough with one or more lipases and / or one or more proteases, b) shaping and / or sheeting the dough, and c) baking the dough, wherein the baked product is selected from the group consisting of biscuits, cookies, rolled versions of biscuits, and crackers.

[0010] In another aspect, the present invention relates to the use of one or more lipases and / or one or more proteases in the production of baked products to reduce the fat and / or sugar used in the recipe for making the baked product.

[0011] In yet another aspect, the present invention relates to reduced-fat baked products obtained by preparing the dough with one or more lipases and / or one or more proteases.

[0012] Sequence Listing Overview SEQ ID NO: 1 is the mature amino acid sequence of 1,3-lipase from Thermomyces lanuginosus.

[0013] SEQ ID NO: 2 is the mature amino acid sequence of a phospholipase from Fusarium oxysporum.

[0014] SEQ ID NO: 3 is the mature amino acid sequence of the phospholipase from Thermomyces lanuginosus.

[0015] SEQ ID NO: 4 is the mature amino acid sequence of the phospholipase from Thermomyces lanuginosus.

[0016] SEQ ID NO: 5 is the mature amino acid sequence of alpha amylase from Aspergillus oryzae.

[0017] SEQ ID NO: 6 is the mature amino acid sequence of the protease from Bacillus amyloliquefaciens.

[0018] SEQ ID NO: 7 is the mature amino acid sequence of xylanase from Thermomyces lanuginosus.

[0019] SEQ ID NO: 8 is the mature amino acid sequence of the protease from Nocardiopsis prasina.

[0020] SEQ ID NO: 9 is the mature amino acid sequence of the protease from Bacillus licheniformis.

[0021] SEQ ID NO: 10 is the mature amino acid sequence of the protease from Fusarium oxysporum.

[0022] SEQ ID NO: 11 is the mature amino acid sequence of the protease from Aspergillus oryzae.

[0023] SEQ ID NO: 12 is the mature amino acid sequence of the protease from Aspergillus oryzae.

[0024] A more complete understanding of the present invention may be obtained by reference to the detailed description and claims when considered in conjunction with the drawings. [Brief explanation of the drawings]

[0025] [Figure 1] FIG. 1 depicts a graph showing the effect of low amounts of lipase on the lateral spread and height coefficients of butter cookies compared to the effect of higher amounts of phospholipase on the cookies. [Figure 2] FIG. 2 shows the texture of butter cookies with a 10% reduction in fat in the negative control and in the lipase test samples. [Figure 3] FIG. 3 shows the texture of butter cookies with a 20% reduction in fat in the negative control and in the lipase test samples. [Figure 4] FIG. 4 shows the slight increase in stack height of butter cookies with a 25% reduction in fat in the positive control and samples tested with lipase and protease. [Figure 5] Figure 5 shows the reduction and hardness of rolled versions of biscuits with a 10% reduction in fat in the positive control and negative control samples for the tests with lipase and protease. [Figure 6] Figure 6 shows the stack height of rolled versions of biscuits with a 10% reduction in fat in the negative control and samples tested with lipase and protease. DETAILED DESCRIPTION OF THE INVENTION

[0026] definition The disclosed embodiments relate to methods, uses and products for preparing reduced fat baked products.

[0027] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Furthermore, terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of this specification and the relevant art, and should not be interpreted in an ideal or overly formal sense unless explicitly defined herein. For the sake of brevity and / or clarity, well-known functions or structures may not be described in detail.

[0028] As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0029] Throughout this disclosure, unless the context requires otherwise, the words "comprise", "comprises" and "comprising" will be understood to mean the inclusion of a stated step or element or group of steps or elements, but not to the exclusion of any other step or element or group of steps or elements.

[0030] The term "consisting of" means including and limited to whatever follows the phrase "consisting of." Thus, the phrase "consisting of" indicates that the listed elements are necessary or mandatory, and that other elements may not be present. The term "consisting essentially of" means including any elements that are listed after the phrase and are limited to other elements that do not interfere with or contribute to the activity or action specified in this disclosure for the listed elements. Thus, the phrase "consisting essentially of" indicates that the listed elements are necessary or mandatory, but that other elements are optional and may or may not be present depending on whether they substantially affect the activity or action of the listed elements.

[0031] As used herein, the term lipase is a 1,3 specific lipase that acts only on triglycerides.

[0032] As used herein, the term reduced fat refers to the use of less fat in the preparation of dough for baked products such as biscuits, cookies, rolled versions of biscuits and / or crackers.

[0033] For the purposes of the present invention, sequence identity between two amino acid sequences is determined as the "longest identity" output using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48:443-453), preferably as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16:276-277), version 6.6.0 or later. The parameters used are a gap opening penalty of 10, a gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. In order for the Needle program to report the longest identity, the no brief option must be specified on the command line. The Needle output labeled "longest identity" is calculated as follows: (identical residues × 100) / (length of alignment−total number of gaps in the alignment).

[0034] For purposes of the present invention, sequence identity between two polynucleotide sequences is preferably determined as the "longest identity" output using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, supra) as implemented in the Needle program of the EMBOSS package (EMBOSS; The European Molecular Biology Open Software Suite, Rice et al., 2000, supra), version 6.6.0 or later. The parameters used are a gap opening penalty of 10, a gap extension penalty of 0.5, and an EDNAFULL (the EMBOSS version in NCBI NUC4.4) substitution matrix. In order for the Needle program to report the longest identity, the no brief option must be specified on the command line. The Needle output labeled "longest identity" is calculated as follows: (identical deoxyribonucleotides × 100) / (length of alignment−total number of gaps in the alignment).

[0035] Several aspects of the present disclosure will hereinafter be described with reference to embodiments thereof; however, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present disclosure as defined by the claims.

[0036] The present invention relates to a method, a use and a product for preparing reduced fat baked products.

[0037] To provide reduced fat in baked products, such as, but not limited to, biscuits, cookies, rolled versions of biscuits, or crackers, the inventors have determined that adding one or more lipases in small amounts (parts per million) while preparing the dough for the baked product and reducing the fat in the dough by 10-25% by substituting flour for the reduced fat provides a baked product with the same or improved baked product qualities or characteristics, such as texture, dough characteristics, size, mouthfeel, balanced flavor / aroma, etc. The one or more lipases act as emulsifiers, which are added to the dough in small amounts compared to other commercially known emulsifiers that must be added in larger amounts (3-5 weight percent). Additionally, the inventors have determined that adding one or more lipases in small amounts while preparing the dough for the baked product and reducing the fat in the dough by 10-25% and sugar by 1-5 weight percent does not compromise the quality or characteristics of the baked product. Furthermore, the inventors have determined that adding a small amount (parts per million) of protease while preparing the dough for a baked product and reducing the fat in the dough by 10-25% by replacing the reduced fat with flour provides the same or improved baked product, and certain qualities or characteristics of the baked product, such as texture, dough characteristics, size, mouthfeel, balanced flavor / aroma, etc. In one aspect, the present invention relates to a method for preparing a baked product, comprising: a) preparing a dough with one or more lipases and / or one or more proteases; b) shaping and / or sheeting the dough; and c) baking the dough. The baked product has at least 10 weight percent (wt%) reduced fat compared to a baked product prepared without the one or more lipases and / or one or more proteases. In another embodiment, the baked product has at least 20 wt% reduced fat compared to a baked product prepared without the one or more lipases and / or one or more proteases. In yet another embodiment, the baked product has at least 25% less fat by weight compared to a baked product prepared without the one or more lipases and / or one or more proteases.The baked product is selected from the group consisting of biscuits, cookies, rolled versions of biscuits, and crackers.

[0038] In one embodiment of the invention, the step of preparing the dough comprises adding one or more sugars, one or more flours, one or more lipases and / or one or more proteases, and one or more fats. The dough comprises 10% to 30%, preferably 15% to 25%, more preferably 20% by weight of the one or more added sugars. In another embodiment, the baked product further comprises reduced sugars compared to a baked product prepared without the one or more lipases and / or one or more proteases. The baked product comprises 1% to 5% reduced sugars compared to a baked product prepared without the one or more lipases and / or one or more proteases.

[0039] In one embodiment of the present invention, the cookie dough comprises 40% to 70% by weight, preferably 55% to 65% by weight, of one or more added flours selected from the group consisting of wheat flour, all-purpose flour, whole wheat flour, rice flour, corn flour, millet flour, soy flour, oat flour, pastry flour, bread flour, gluten-free flour, tapioca flour, and self-rising flour.

[0040] In one embodiment of the present invention, the cookie dough comprises 0.00004% to 0.0002% by weight, preferably 0.00006% to 0.0001% by weight of one or more added lipases.

[0041] In one embodiment of the present invention, the dough contains one or more added lipases at 0.4 to 2 milligrams (mg) of enzyme protein / kilogram (kg) of flour, preferably 0.6 to 1 mg of enzyme protein / kg of flour.

[0042] In one embodiment of the present invention, the cookie dough contains 20 parts per million (ppm) to 100 ppm, preferably 25 ppm to 55 ppm, of one or more added lipases. The lipase is a Thermomyces lanuginosus (formerly Humicola lanuginosus) wild-type lipase. In a preferred embodiment, the lipase is a 1,3-specific lipase that acts only on triglycerides.

[0043] In one embodiment of the present invention, the dough for the rolled variant of biscuits comprises 0.00001% to 0.0006% by weight, preferably 0.000014% to 0.000016% by weight of one or more added lipases.

[0044] In one embodiment of the present invention, the dough for the rolling variant of biscuits comprises one or more added lipases at 0.1 to 0.6 milligrams (mg) of enzyme protein / kilogram (kg) of flour, preferably 0.14 to 0.16 mg of enzyme protein / kg of flour.

[0045] In one embodiment of the present invention, the dough for the rolled version of the biscuit comprises between 5 parts per million (ppm) and 30 ppm of one or more added lipases.

[0046] In one embodiment of the invention, the one or more lipases are polypeptides selected from the group consisting of: (a) a polypeptide having at least 80% sequence identity to SEQ ID NO: 1; (b) a polypeptide derived from SEQ ID NO: 1 by having 1 to 30 modifications, such as substitutions, deletions and / or insertions, at one or more positions, such as 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 modifications, particularly substitutions; (c) a polypeptide derived from the polypeptide of (a), wherein the N-terminus and / or C-terminus is extended by the addition of one or more amino acids; and (d) A fragment of the polypeptide of (a).

[0047] In another embodiment, the polypeptide has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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%, at least 99% or 100% sequence identity to the mature polypeptide coding sequence of SEQ ID NO:1.

[0048] In a preferred embodiment of the invention, the lipase is a polypeptide comprising, consisting essentially of or consisting of SEQ ID NO:1.

[0049] In one embodiment, the polypeptide may have an N-terminal and / or C-terminal extension of one or more amino acids, for example 1 to 5 amino acids.

[0050] In another aspect, the polypeptide is derived from SEQ ID NO: 1 by substitution, deletion, or addition of one or several amino acids. In another aspect, the polypeptide is derived from the mature polypeptide of SEQ ID NO: 1 by substitution, deletion, or addition of one or several amino acids. In some embodiments, the polypeptide is a variant of SEQ ID NO: 1 comprising substitutions, deletions, and / or insertions at one or more positions. In one embodiment, the number of amino acid substitutions, deletions, and / or insertions introduced into the polypeptide of SEQ ID NO: 1 is 15 or less, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. The amino acid changes may be minor in nature, i.e., conservative amino acid substitutions or insertions that have little effect on protein folding and / or activity; small deletions, typically 1-30 amino acids; small amino- or carboxy-terminal extensions such as an amino-terminal methionine residue; small linker peptides of 20-25 residues or less; or small extensions that facilitate purification by altering net charge or another function, e.g., a polyhistidine tract, an antigenic epitope, or a binding module.

[0051] In one embodiment of the invention, the dough comprises 12% to 35% by weight of one or more fats selected from the group consisting of palm oil, palm olein, butter, clarified butter, anhydrous milk fat, corn oil, margarine, olive oil, hydrogenated vegetable fat, sunflower oil, rapeseed oil, rice bran oil, lard, soybean oil and eggs. In a preferred embodiment, the fat is palm oil or rapeseed oil.

[0052] In one embodiment, the steps for preparing the dough include: i. 10% to 30% by weight of one or more sugars; ii. 40% to 70% by weight of one or more flowers; iii. 0.2% to 2% by weight of one or more lipases; and iv. Including adding 12% to 35% by weight of one or more fats.

[0053] In one embodiment, the steps for preparing the dough include: i. 10% to 30% by weight of one or more sugars; ii. 40% to 70% by weight of one or more flowers; iii. 20 ppm to 100 ppm of one or more lipases; and iv. Including adding 12% to 35% by weight of one or more fats.

[0054] In one embodiment, the step for preparing the dough comprises adding one or more proteases.

[0055] In one embodiment of the present invention, the cookie dough comprises 0.0.00015% to 0.003% by weight, preferably 0.00003% to 0.0006% by weight of one or more added proteases.

[0056] In one embodiment of the present invention, the cookie dough contains one or more added proteases at 1.5 to 30 milligrams (mg) of enzyme protein / kilogram (kg) of flour, preferably 0.3 to 6 mg of enzyme protein / kg of flour.

[0057] In one embodiment of the present invention, the cookie dough comprises between 5 parts per million (ppm) and 100 ppm of one or more added proteases.

[0058] In one embodiment of the present invention, the dough for rolled modified biscuits comprises 0.00015% to 0.009% by weight, preferably 0.00003% to 0.00018% by weight of one or more added proteases.

[0059] In one embodiment of the present invention, the dough for rolled modified biscuits comprises one or more added proteases at 1.5 to 90 milligrams (mg) of enzyme protein / kilogram (kg) of flour, preferably 0.3 to 18 mg of enzyme protein / kg of flour.

[0060] In one embodiment of the present invention, the dough for rolled modified biscuits comprises between 5 parts per million (ppm) and 300 ppm of one or more added proteases.

[0061] In one embodiment of the present invention, the one or more proteases are derived from Bacillus amyloliquefaciens.

[0062] In one embodiment of the invention, the one or more proteases are polypeptides selected from the group consisting of: (a) a polypeptide having at least 80% sequence identity to SEQ ID NO:6; (b) a polypeptide derived from SEQ ID NO: 6 by having 1 to 30 modifications, such as substitutions, deletions and / or insertions, at one or more positions, such as 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 modifications, particularly substitutions; (c) a polypeptide derived from the polypeptide of (a), wherein the N-terminus and / or C-terminus is extended by the addition of one or more amino acids; and (d) A fragment of the polypeptide of (a).

[0063] In another embodiment, the polypeptide has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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%, at least 99% or 100% sequence identity to the mature polypeptide coding sequence of SEQ ID NO:6.

[0064] In a preferred embodiment of the invention, the protease is a polypeptide comprising, consisting essentially of, or consisting of SEQ ID NO:6.

[0065] In one embodiment, the polypeptide may have an N-terminal and / or C-terminal extension of one or more amino acids, for example 1 to 5 amino acids.

[0066] In another aspect, the polypeptide is derived from SEQ ID NO: 6 by substitution, deletion, or addition of one or more amino acids. In another aspect, the polypeptide is derived from the mature polypeptide of SEQ ID NO: 6 by substitution, deletion, or addition of one or more amino acids. In some embodiments, the polypeptide is a variant of SEQ ID NO: 6 comprising substitutions, deletions, and / or insertions at one or more positions. In one embodiment, the number of amino acid substitutions, deletions, and / or insertions introduced into the polypeptide of SEQ ID NO: 6 is 15 or less, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. The amino acid changes may be minor in nature, i.e., conservative amino acid substitutions or insertions that have little effect on protein folding and / or activity; small deletions, typically 1-30 amino acids; small amino- or carboxy-terminal extensions such as an amino-terminal methionine residue; small linker peptides of 20-25 residues or less; or small extensions that facilitate purification by altering net charge or another function, e.g., a polyhistidine tract, an antigenic epitope, or a binding module.

[0067] In a further embodiment, the step of preparing the dough comprises adding a combination of one or more lipases and one or more proteases.

[0068] In another embodiment, the step of preparing the dough includes adding one or more leavening agents, wherein the dough comprises 0.3% to 3% by weight of one or more added leavening agents, the leavening agents being selected from the group consisting of sodium bicarbonate, ammonium bicarbonate, disodium dihydrogen pyrophosphate (SAPP), sodium phosphate, monocalcium phosphate, sodium aluminum sulfate, potassium bitartrate, sodium aluminum phosphate, glucono-delta-lactone, and adipic acid, and combinations thereof.

[0069] In yet another embodiment, the step of preparing the dough further comprises adding other ingredients, such as invert syrup, salted or unsalted butter, lecithin, milk powder, distilled mono- and diglycerides (DMG), salt, liquid glucose, cheese, fresh milk, colorants, flavorings, sodium metabisulfate, sweetened condensed milk, spices and seasonings, glycerol monostearate (GMS), diacetyl tartaric acid esters of mono- and diglycerides of fatty acids (DATEM), and / or water.

[0070] In one embodiment, the steps for preparing dough include creaming, mixing, and maintaining a suitable temperature for the dough. One or more flours and one or more lipases or / and one or more proteases are added during the mixing step while preparing the dough. If one or more lipases or / and one or more proteases are added during mixing, no resting period is required. The present invention improves dough properties such as dough pH, ​​dough texture, dough temperature, and dough sheeting.

[0071] In one embodiment, the dough is formed in one or more dough molds, hi another embodiment, the dough is sheeted in one or more sheeters.

[0072] In one embodiment, the dough is cut with one or more cutters after being sheeted / rolled.

[0073] In another embodiment, the sheeted dough is rolled and folded one or more times for rolled versions of biscuits.

[0074] In one embodiment, the dough is baked in an oven selected from the group consisting of a tunnel oven, a rotary oven, and a deck oven. The present invention improves the dimensions of the baked product, e.g., cookies. The dimensions can be weight, stack height, average length, average width, volume, density, lateral spread factor, height factor, etc.

[0075] In another aspect, the present invention relates to the use of one or more lipases and / or one or more proteases in the production of baked products to reduce the fat and / or sugars used in preparing the baked product. One or more properties of the baked product are maintained after reducing the fat and / or sugars while preparing the dough for the baked product. The one or more properties are selected from the group consisting of mouthfeel, texture, taste, bite, spreadability, color-height-length, volume, moisture, strength, hardness, crispness, and shelf life. The dough of the baked product has at least 10% less fat by weight compared to a baked product prepared without the one or more lipases and / or one or more proteases.

[0076] In another aspect, the present invention relates to a baked product obtainable by the method defined in the above embodiments or obtained by the use of one or more lipases or / and one or more proteases.

[0077] Particular embodiments of the present disclosure are described in the following numbered sections: 1. A method for preparing a reduced fat baked product comprising: a. At least: i. one or more sugars; ii. one or more flowers; iii. one or more lipases or / and one or more proteases; and iv. preparing the dough by adding one or more fats; b. shaping the dough in one or more dough molds and / or sheeting the dough in one or more dough sheeters; c. baking the dough; The method, wherein the baked product has at least 10% by weight fat reduction when compared to a baked product prepared without the one or more lipases and / or one or more proteases. 2. The method for preparing a baked product according to item 1, wherein the baked product is selected from the group consisting of biscuits, cookies, rolled versions of biscuits, and crackers. 3. The method for preparing a baked product according to paragraph 1 or 2, wherein the dough comprises 10% to 30% by weight of one or more added sugars. 4. The method for preparing a baked product according to any one of items 1 to 3, wherein the flour is selected from the group consisting of wheat flour, all-purpose flour, whole wheat flour, rice flour, corn flour, millet flour, soy flour, oat flour, pastry flour, bread flour, gluten-free flour, tapioca flour, and self-rising flour. 5. A method for preparing a baked product according to any one of paragraphs 1 to 4, wherein the dough comprises 40% to 70% by weight of one or more added flours. 6. A method for preparing a baked product according to any one of paragraphs 1 to 5, wherein the lipase is a wild-type lipase of Thermomyces lanuginosus (formerly known as Humicola lanuginosus). 7. The method for preparing a baked product according to any one of paragraphs 1 to 6, wherein the one or more lipases are polypeptides selected from the group consisting of: (a) a polypeptide having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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%, at least 99% or 100% sequence identity to SEQ ID NO:1; (b) a polypeptide derived from SEQ ID NO: 1 by having 1 to 30 modifications, such as substitutions, deletions and / or insertions, at one or more positions, such as 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 modifications, particularly substitutions; (c) a polypeptide derived from the polypeptide of (a), wherein the N-terminus and / or C-terminus are extended by the addition of one or more amino acids; and (d) A fragment of the polypeptide of (a). 8. A method for preparing a baked product according to any one of items 1 to 7, wherein the one or more lipases are polypeptides having the sequence of SEQ ID NO: 1. 9. The method for preparing a baked product according to any one of items 1 to 8, wherein the cookie dough comprises 0.00001% to 0.0002% by weight, preferably 0.00006% to 0.0001% by weight, of one or more added lipases. 10. A method for preparing a baked product according to any one of paragraphs 1 to 9, wherein the dough for the rolled variant of the biscuit comprises 0.00001% to 0.0006% by weight, preferably 0.000014% to 0.000016% by weight, of one or more added lipases. 11. A method for preparing a baked product according to any one of paragraphs 1 to 10, wherein the cookie dough contains one or more added lipases in an amount of 0.4 to 2 mg enzyme protein / kg flour, preferably 0.6 to 1 mg enzyme protein / kg flour. 12. A method for preparing a baked product according to any one of paragraphs 1 to 11, wherein the dough for the biscuit rolling variant contains one or more added lipases in an amount of 0.1 to 0.6 mg enzyme protein / kg flour, preferably 0.14 to 0.16 mg enzyme protein / kg flour. 13. The method for preparing a baked product according to any one of paragraphs 1 to 12, wherein the dough comprises between 20 parts per million (ppm) and 100 ppm of one or more added lipases. 14. A method for preparing a baked product according to any one of paragraphs 1 to 13, wherein the dough for the rolled variant of the biscuit comprises between 5 parts per million (ppm) and 30 ppm of one or more added lipases. 15. A method for preparing a baked product according to any one of paragraphs 1 to 14, wherein the step of preparing the dough includes adding one or more proteases. 16. A method for preparing a baked product according to any one of items 1 to 15, wherein the protease is derived from Bacillus amyloliquefaciens. 17. The method for preparing a baked product according to any one of paragraphs 1 to 16, wherein the one or more proteases are polypeptides selected from the group consisting of: (a) a polypeptide having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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%, at least 99% or 100% sequence identity to SEQ ID NO:6; (b) a polypeptide derived from SEQ ID NO: 6 by having 1 to 30 modifications, such as substitutions, deletions and / or insertions, at one or more positions, such as 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 modifications, particularly substitutions; (c) a polypeptide derived from the polypeptide of (a), wherein the N-terminus and / or C-terminus are extended by the addition of one or more amino acids; and (d) A fragment of the polypeptide of (a). 18. A method for preparing a baked product according to any one of paragraphs 1 to 17, wherein the one or more proteases are polypeptides having the sequence of SEQ ID NO:6. 19. A method for preparing a baked product according to any one of items 1 to 18, wherein the cookie dough contains 0.00015% to 0.003% by weight, preferably 0.00003% to 0.0006% by weight, of one or more proteases. 20. A method for preparing a baked product according to any one of paragraphs 1 to 19, wherein the dough for the rolled variant of the biscuit contains 0.00015% to 0.009% by weight, preferably 0.00003% to 0.00018% by weight, of one or more proteases. 21. A method for preparing a baked product according to any one of paragraphs 1 to 20, wherein the cookie dough contains one or more added proteases in an amount of 1.5 to 30 mg enzyme protein / kg flour, preferably 0.3 to 0.6 mg enzyme protein / kg flour. 22. A method for preparing a baked product according to any one of paragraphs 1 to 21, wherein the dough for the biscuit rolling variant contains one or more added proteases in an amount of 1.5 to 90 mg enzyme protein / kg flour, preferably 0.3 to 18 mg enzyme protein / kg flour. 23. The method for preparing a baked product according to any one of paragraphs 1 to 22, wherein the cookie dough comprises between 5 parts per million (ppm) and 100 ppm of one or more added proteases. 24. A method for preparing a baked product according to any one of paragraphs 1 to 23, wherein the dough for a rolled version of a biscuit comprises between 5 parts per million (ppm) and 300 ppm of one or more added proteases. 25. A method for preparing a baked product according to any one of paragraphs 1 to 24, wherein the step of preparing the dough comprises adding a combination of one or more lipases and one or more proteases. 26. A method for preparing a baked product according to any one of items 1 to 25, wherein the fat is selected from the group consisting of palm oil; palm olein, butter, clarified butter, anhydrous milk fat, corn oil, margarine, olive oil, hydrogenated vegetable fat, sunflower oil, rapeseed oil, rice bran oil, lard, soybean oil, and eggs. 27. A method for preparing a baked product according to any one of paragraphs 1 to 26, wherein the fat is palm oil. 28. A method for preparing a baked product according to any one of paragraphs 1 to 27, wherein the dough comprises 12% to 35% by weight of one or more fats. 29. The steps for preparing the dough are: i. 10% to 30% by weight of one or more sugars; ii. 40% to 70% by weight of one or more flowers; iii. 0.00001% to 0.0002% by weight of one or more lipases and / or one or more proteases; and iv. A method for preparing a baked product according to any one of paragraphs 1 to 28, comprising adding 12% to 35% by weight of one or more fats. 30. The steps for preparing the dough are: i. 10% to 30% by weight of one or more sugars; ii. 40% to 70% by weight of one or more flowers; iii. 0.1-2 mg of enzyme protein / kg of flour of one or more added lipases and / or one or more proteases; and iv. A method for preparing a baked product according to any one of paragraphs 1 to 29, comprising adding 12% to 35% by weight of one or more fats. 31. The steps for preparing the dough are: i. 10% to 30% by weight of one or more sugars; ii. 40% to 70% by weight of one or more flowers; iii. 5 ppm to 300 ppm of one or more lipases and / or one or more proteases; and iv. A method for preparing a baked product according to any one of paragraphs 1 to 30, comprising adding 12% to 35% by weight of one or more fats. 32. A method for preparing a baked product according to any one of paragraphs 1 to 31, wherein the method further reduces sugars in the baked product compared to a baked product prepared without one or more lipases and / or one or more proteases. 33. The method for preparing a baked product according to paragraph 24, wherein the baked product has 1% to 5% less sugar by weight compared to a baked product prepared without one or more lipases and / or one or more proteases. 34. A method for preparing a baked product according to any one of paragraphs 1 to 33, wherein the baked product has at least 20% by weight less fat compared to a baked product prepared without one or more lipases and / or one or more proteases. 35. A method for preparing a baked product according to any one of paragraphs 1 to 34, wherein the baked product has at least 25% by weight less fat compared to a baked product prepared without one or more lipases and / or one or more proteases. 36. A method for preparing a baked product according to any one of paragraphs 1 to 35, wherein the step of preparing the dough further comprises adding one or more leavening agents. 37. A method for preparing a baked product according to paragraph 36, wherein the leavening agent is selected from the group consisting of sodium bicarbonate, ammonium bicarbonate, disodium dihydrogen pyrophosphate (SAPP), sodium phosphate, monocalcium phosphate, sodium aluminum sulfate, potassium bitartrate, sodium aluminum phosphate, glucono-delta-lactone, and adipic acid, and combinations thereof. 38. A method for preparing a baked product according to paragraphs 36-37, wherein the dough comprises 0.3% to 3% by weight of one or more added leavening agents. 39. The method for preparing a baked product according to any one of paragraphs 1 to 38, wherein the step of preparing the dough further comprises adding invert syrup, salted or unsalted butter, lecithin, milk powder, distilled mono- and diglycerides (DMG), salt, liquid glucose, cheese, fresh milk, colorants, flavorings, sodium metabisulfate, sweetened condensed milk, spices and seasonings, glycerol monostearate (GMS), diacetyl tartaric acid esters of mono- and diglycerides of fatty acids (DATEM), and / or water. 40. Use of one or more lipases in the production of baked products to reduce the fat and / or sugar used in the preparation of the baked product. 41. The use according to paragraph 40, wherein one or more properties of the baked product are maintained. 42. Use according to paragraph 40 or 41, wherein the one or more properties are selected from the group consisting of mouthfeel, texture, taste, chewiness, spreadability, color-height-length, volume, moisture, strength, hardness, crispness and shelf life. 43. Use according to any one of paragraphs 40 to 42, wherein the baked product has at least 10% by weight less fat, preferably at least 20% by weight less fat, even more preferably at least 25% by weight less fat, compared to a baked product prepared without the one or more lipases and / or one or more proteases. 44. A baked product obtainable by the method according to any one of paragraphs 1 to 39 or by the use according to any one of paragraphs 40 to 43. [Example]

[0078] The following examples are not intended to be a detailed list of all the various ways in which the present disclosure may be implemented or all the features that may be added to the present disclosure. Those skilled in the art will appreciate that many modifications and additions to the various embodiments may be made without departing from the present disclosure. Therefore, the following description is intended to illustrate certain specific embodiments of the present invention, and is not intended to exhaustively identify all modifications, combinations, and variations thereof.

[0079] Unless otherwise indicated, percentages set forth in the following examples are by weight based on the total weight of the composition.

[0080] Materials and Methods The experiment was carried out in Denmark using standard ingredients for making cookies in Denmark. The standard ingredients and procedures listed in Example 1 below were used to prepare the cookies. Enzymes used in the study:

[0081] [Table 1]

[0082] [Table 2]

[0083] Example 1 - Process Flow for Preparing Butter Cookies 1. Creaming: Rapeseed oil, invert syrup, salted butter, lecithin, distilled mono- and diglyceride (DMG) paste, and flavors were weighed into a mixing bowl and mixed at medium speed for 2 minutes. Sugar and nonfat milk powder (SMP) were added to the mixing bowl and mixed at medium speed for 3 minutes. Salt, sodium bicarbonate, and ammonium bicarbonate were dissolved in water and added to the mixing bowl. The ingredients in the mixing bowl were mixed at medium speed for 5 minutes. 2.Mixing: Flour was added and the ingredients were mixed for 5-6 minutes for proper dough consistency. Water was added as per requirement to adjust the dough consistency. 3.Temperature: The temperature of the dough was maintained at 23-25 ​​degrees Celsius (°C). 4. Forming / sheeting: The dough was shaped using a butter cookie cutter and sheeted to a thickness of 3.5 millimeters (mm) using a dough sheeter. 5. Baking: The formed and sheeted dough was baked at 235°C for 6 to 7 minutes. 6. Cooling and packaging: The cookies were cooled for 1.5 times the baking time and wrapped in aluminum foil.

[0084] Example 2 - Lipase screening: Two lipases were tested to achieve a 20% reduction in fat. 1. Lipase - 1,3-specific lipase that acts only on triglycerides (SEQ ID NO: 1) 2. Phospholipases: Phospholipase 1 (SEQ ID NO: 2), Phospholipase 2 (SEQ ID NO: 3) and Phospholipase 3 (SEQ ID NO: 4) acting on triglycerides and polar lipids.

[0085] Lipase was added with the flour during the mixing step of the process of Example 1.

[0086] [Table 3]

[0087] [Table 4]

[0088] Tables 1 and 2 show that lipase (SEQ ID NO: 1) at 10 ppm performed better in cookie stack height when compared to phospholipases (SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4).

[0089] The sensory characteristics of cookies baked using lipase were scored higher when compared to cookies baked using phospholipase. Figure 1 shows that lipase provides similar or better results at a very low dosage of 10 ppm compared to phospholipase. The lateral spread and height coefficients of cookies with 20% reduced fat and 10 ppm lipase are similar or better than those of control cookies and cookies baked using 50 ppm or more of phospholipase. Therefore, using lipase provides a cost-effective approach to baking cookies.

[0090] Example 3 - Process Flow for Preparing 10-20% Reduced Fat Butter Cookies Butter cookies were prepared using the process steps defined in Example 1. The amounts of ingredients used in the process for preparing the butter cookies are set forth in Table 3 (10% fat reduction) and Table 4 (20% fat reduction).

[0091] [Table 5]

[0092] [Table 6]

[0093] 1. Creaming: Three samples were prepared: a positive control, a negative control and a test with lipase (SEQ ID NO: 1). 2.Mixing: Wheat flour was added to each of the three samples. Lipase (SEQ ID NO: 1) was added only to the sample tested with lipase. No dough resting time was required. 3.Temperature: The temperature of the dough was maintained at 23-25 ​​degrees Celsius (°C). 4. Forming / sheeting: The dough was shaped using a butter cookie cutter and sheeted to a thickness of 3.5 millimeters (mm) using a dough sheeter.

[0094] [Table 7]

[0095] [Table 8]

[0096] The dough properties of the lipase-tested samples were similar or improved compared to the positive control. The lipase-tested doughs were better in machinability than the control dough at 10 and 20% fat reduction, while the fat-reduced dough without lipase (negative control) was very dry and coarse during the sheeting / forming process. Thus, lipase exhibits emulsifying activity in flour instead of fat reduction. 5. Baking: The sheeted and shaped dough was baked at 235°C for 6-7 minutes. 6. Cooling and packaging: The cookies were cooled for 1.5 times the baking time and wrapped in aluminum foil.

[0097] [Table 9]

[0098] [Table 10]

[0099] The dimensions of the sample-lipase test butter cookies were similar or improved compared to the positive control sample butter cookies.

[0100] Figures 2 and 3 show that the positive control and lipase test samples have similar textures. Therefore, a 10% and 20% reduction in fat in the butter cookie baking process using lipase does not impair the texture of the butter cookies, as shown in Figures 2 and 3.

[0101] Lipase test cookies show a significant increase in cookie stack height at both 10% and 20% fat reduction when compared to the positive and negative controls (fat reduction without lipase). The density of lipase test cookies was also less than the controls.

[0102] Sugars can also be reduced in cookies with higher lipase dosages, as the emulsifying properties of lipase increase the perception of sugars in the cookies. Therefore, sugars were reduced by 2% and substituted for wheat flour along with a 20% fat reduction with a 50 ppm dose of lipase.

[0103] Example 4 - 3-point test method-1 A total of 15 panelists were tasters using a triangle test. Three samples (two identical and one different) were presented to each panelist simultaneously, and each panelist was then asked to identify the one that was different from the other two. Below are the six ways in which the samples were presented to the panelists, and each sample was coded with a three-digit number:

[0104] [Table 11]

[0105] Panelists were asked to identify the code on the scorecard that they believed to be the odd sample. This method requires panelists to choose between samples. The decision is based on the minimum number of correct responses required for significance at a pre-determined significance level, taking into account the total number of responses obtained. The goal was to identify the odd sample in a triangle test.

[0106] [Table 12]

[0107] [Table 13]

[0108] None of the panelists noticed or noted a firmer texture in the cookies with fat reduction and corresponding increase in flour percentage in the sample-lipase test. Thus, lipase maintains the texture of the cookies even after 10% and 20% fat reduction during baking.

[0109] Panelists felt that the flavor / aroma in the control sample was unbalanced, while the test flavor / aroma was well rounded / balanced.

[0110] For the 10% fat reduction, few panelists noted that the test samples were slightly sweeter than the control, which was due to the emulsifying properties of the lipase, and the increased sweetness is a positive attribute as it also improves the mouthfeel and overall satisfaction of the cookie.

[0111] For the 20% fat reduction, 2% sugar reduction, and wheat flour replacement, few panelists (2 / 15) noted that the control sample was slightly sweeter than the test. Thus, lipase also compensates for sugar reduction as well as fat reduction at high doses.

[0112] Example 5 - Screening of lipases and other enzymes for 25% fat and 2% sugar reduction in butter cookies The following tests in Table 11 were performed to evaluate the synergistic effects of lipase with other enzymes.

[0113] [Table 14]

[0114] [Table 15]

[0115] The cookie sample with the combination of Lipase and Protease 1 had a similar texture to the positive control at 25% fat reduction, so this combination was carried forward for further testing.

[0116] Example 6 - Process Flow for Preparing 25% Reduced Fat Butter Cookies Butter cookies were prepared using the process steps defined in Example 1. The amounts of ingredients used in the process to prepare the butter cookies are listed in Table 12 (25% fat reduction).

[0117] [Table 16]

[0118] 1. Creaming: Three samples were prepared: a positive control, a test with lipase and test 1 with lipase and protease. 2.Mixing: Wheat flour was added to each of the three samples. Lipase was added only to the test with Sample-Lipase. Lipase and Protease 1 were added only to the test with Sample-Lipase and Protease 1. No resting time was required for the dough. 3.Temperature: The temperature of the dough was maintained at 23-25 ​​degrees Celsius (°C). 4. Forming / sheeting: The dough was shaped using a butter cookie cutter and sheeted to a thickness of 3.5 millimeters (mm) using a dough sheeter.

[0119] [Table 17]

[0120] The dough properties of the samples tested with Lipase and Protease 1 were similar or improved compared to the positive control sample. Lipase and Protease 1 therefore exhibit emulsifying activity in wheat flour instead of fat reduction. 5. Baking: The sheeted and shaped dough was baked at 235°C for 6-7 minutes. 6. Cooling and packaging: The cookies were cooled for 1.5 times the baking time and wrapped in aluminum foil.

[0121] [Table 18]

[0122] A 25% fat reduction with a 60 ppm lipase dose results in a significant reduction in stack height. The test samples with 60 ppm lipase had a harder texture and a more chalky mouthfeel than the positive control sample.

[0123] Cookie shrinkage and hard cookie texture revealed that dough resting was required when the fat was reduced at higher percentages. To address the cookie shrinkage and hard texture, another enzyme (Protease 1) was used to achieve a texture similar to that of the positive control.

[0124] [Table 19]

[0125] The dimensions of the butter cookies from the test samples using Lipase and Protease 1 were similar or improved compared to the butter cookies from the positive control sample.

[0126] Figure 4 shows that the samples from the positive control and the test using Lipase and Protease 1 have similar textures. Therefore, a 25% fat reduction in the butter cookie baking process using Lipase and Protease 1 does not impair the texture of the butter cookies, as shown in Figure 4. The test samples also have a slightly increased stack height than the positive control.

[0127] The test cookies with Lipase and Protease 1 show a significant increase in cookie stack height at 25% fat reduction when compared to the positive control. The density of the test cookies with Lipase and Protease 1 was also lower than the positive control.

[0128] Sugars can also be reduced in cookies with higher dosages of lipase and protease 1, as the emulsifying properties of lipase and protease 1 increase the perception of sugars in the cookies. Therefore, sugars were reduced by 2% and substituted for wheat flour with a 25% fat reduction at a dosage of 50 ppm lipase and 8 ppm protease 1.

[0129] Example 7 - 3-Point Test Method 2 A total of 20 panelists were tasters for the triangle test. The triangle test was performed in the same manner as in Example 4 using the butter cookie samples of Example 6.

[0130] [Table 20]

[0131] None of the panelists noticed or noted a firmer texture in the cookies due to the fat reduction and corresponding increase in flour percentage in the samples tested with Lipase and Protease 1. Most panelists preferred the test samples over the control due to the improved texture.

[0132] Example 8 - Process flow for preparing rolled / hard dough variants of biscuits 1. Creaming: The fat, invert syrup, glycerol monostearate (GMS), DATEM, and flavor were weighed into a mixing bowl and mixed at medium speed for 2 minutes. The sugar and nonfat milk powder (SMP) were added to the mixing bowl and mixed at medium speed for 3 minutes. The salt, sodium bicarbonate, and ammonium bicarbonate were dissolved in water and added to the mixing bowl. The ingredients in the mixing bowl were mixed at medium speed for 5 minutes. 2.Mixing: Wheat flour was added to the mixing bowl. SMBS was added to the mixing bowl in the form of a solution in water. The ingredients were mixed for 5-6 minutes for proper dough consistency. Water was added as per requirement to adjust the dough consistency. 3.Temperature: The temperature of the dough was maintained at 40-42 degrees Celsius (°C). 4.Resting time The dough was allowed to rest for 20 minutes. 5. Sheeting: After the dough resting period, it is rolled out on a sheeter and the dough sheet is folded. The rolling and folding process is repeated six times. The thickness of the final sheet after rolling is 2 millimeters (mm). 6.Cutting: The rolled dough sheets were cut with a Teflon cutter with a dhoker pinhole. 7. Baking: After cutting, the dough sheets were baked at a temperature of 270°C for 5-6 minutes. 8. Cooling and packaging: The cookies were cooled for 1.5 times the baking time and wrapped in aluminum foil.

[0133] Example 9 - Lipase for 10% fat reduction in biscuit rolling / hard dough variants Rolled / hard dough versions of cookies were prepared using the process steps defined in Example 8. Tests were conducted using lipase alone to reduce the fat of rolled / hard dough versions of biscuits. Initial tests were designed to achieve a 10% fat reduction in the rolled / hard dough version using 30 ppm lipase alone.

[0134] [Table 21]

[0135] Results: A decrease in volume was observed in the test samples and an increase in density.

[0136] Sensory evaluation: The test samples were drier compared to the positive control sample. The reduced fat samples were noted to have a firmer texture when compared to the positive control sample.

[0137] Conclusion: The reduction in biscuits and harder texture highlights the need for dough resting when the fat is reduced to a higher percentage while making rolled / hard dough variants of biscuits. Therefore, other enzymes (proteases) had to be used to achieve a texture similar to that of the positive control.

[0138] Example 10 - Protease screening for 10% fat reduction in rolled / hard dough variants of biscuits Rolled / hard dough versions of biscuits were prepared using the process steps defined in Example 8. The following eight proteases were tested for a 10% fat reduction in the rolled / hard dough versions of biscuits: 1. Protease 1 2. Blend 1 3. Protease 2 4. Blend 2 5. Protease 3 6. Protease 4 7. Blend 3 8. Blend 4 Details of protease screening are as follows: 1. The positive control had 10.64% total fat on a finished product (FG) basis. 2. The negative control was a 10% fat reduction at 9.35% fat percentage on an FG basis without enzyme. 3. Tests were conducted using various proteases to achieve a 10% fat reduction at 9.35% fat percentage on an FG basis.

[0139] [Table 22]

[0140] [Table 23]

[0141] [Table 24]

[0142] [Table 25]

[0143] All proteases had the same or better dough properties, biscuit size, and sensory attributes compared to the positive control. Biscuits made with Protease 1 had better dough properties, biscuit size, and sensory attributes when compared to other types of proteases. Protease 1 worked at very low dosages, making it a candidate for further testing with lipases.

[0144] Example 11 - Process flow for preparing a rolled / hard dough variant of a 10% reduced fat biscuit Rolled / hard dough variants of biscuits were prepared using the process steps defined in Example 8. Tests were carried out using Lipase and Protease 1 with a 10% reduction in fat.

[0145] [Table 26]

[0146] [Table 27]

[0147] Results: Tests using Protease 1 and Lipase were found to have better spread and stack height coefficients as well as the positive control.

[0148] Protease 1 alone achieved a volume and density similar to that of the positive control, but the sensory characteristics indicated a dry mouthfeel. Therefore, lipase was added to achieve a mouthfeel similar to that of the positive control. The mouthfeel was similar with the combination of Protease 1 and lipase because lipase enhances emulsifying properties.

[0149] Example 12 - Process flow for preparing a rolled / hard dough variant of a 10% reduced fat biscuit A rolled / hard dough variant of the biscuit was prepared using the process steps defined in Example 8. The amounts of ingredients used in the process to prepare such biscuits are set out in Table 20 (10% fat reduction in the rolled variant).

[0150] [Table 28]

[0151] 1. Creaming: Three samples were prepared: a positive control, a negative control and test 1 with lipase and protease. 2.Mixing: Wheat flour was added to each of the three samples. Lipase and Protease 1 were added only to the sample - test with Lipase and Protease 1. 3.Temperature: The temperature of the dough was maintained at 40-42 degrees Celsius (°C). The dough was allowed to rest for 20 minutes. 4. Sheeting: The dough was rolled on a sheeter and the dough sheet was folded. The rolling and folding process was repeated six times. The final sheet thickness after rolling was 2 millimeters (mm).

[0152] [Table 29]

[0153] The test dough with Lipase and Protease 1 performed better than the control dough in terms of machineability at 10% fat reduction, and the fat-reduced dough without Lipase and Protease 1 (negative control) dried out and shrunk during the sheeting process, indicating that Lipase and Protease 1 have emulsifying and dough-resting activity in flour instead of fat reduction. 5.Cutting: The rolled dough sheets were cut with a Teflon cutter equipped with a Doker pinhole. 6. Baking: The rolled dough sheets were baked at a temperature of 270°C for 5-6 minutes. 7. Cooling and packaging: The crackers were cooled for 1.5 times the baking time and wrapped in aluminum foil.

[0154] [Table 30]

[0155] The cracker dimensions of the samples - Lipase and Protease 1 tests were similar or improved compared to the positive control sample crackers.

[0156] Figures 5 and 6 show that the positive control and the test with Lipase and Protease 1 sample have similar textures. Therefore, a 10% fat reduction in the rolling variant of the biscuit baking process using Lipase and Protease 1 does not impair the texture of the cracker as shown in Figures 5 and 6.

[0157] The test crackers with lipase and protease 1 show a slight increase in cracker stack height at both 10% fat reduction when compared to the positive control and the negative control (fat reduction without lipase and protease 1). The density of the test crackers with lipase and protease 1 was also lower than the control.

[0158] Example 13 - 3-Point Test Method 3 A total of 20 panelists served as tasters for the triangle test, which was performed in the same manner as in Example 4 using the cracker samples of Example 8.

[0159] [Table 31]

[0160] None of the panelists noticed or noted a harder texture in the crackers with the 10% fat reduction and corresponding increase in flour percentage in the test with Sample-Lipase and Protease 1.

Claims

1. 1. A method for preparing a reduced fat baked product comprising: a. At least: i. one or more sugars; ii. one or more flowers; iii. one or more lipases or / and one or more proteases; and iv. preparing the dough by adding one or more fats; b. shaping the dough in one or more dough molds and / or sheeting the dough in one or more dough sheeters; c. baking the dough; The baked product has at least 10 weight percent (wt%) to 25 wt% reduction in fat when compared to a baked product prepared without the one or more lipases and / or one or more proteases.

2. 10. The method for preparing a baked product of claim 1, wherein the baked product is selected from the group consisting of biscuits, cookies, rolled versions of biscuits, and crackers.

3. 3. A method for preparing a baked product according to claim 1 or 2, wherein the dough comprises from 10% to 30% by weight of one or more added sugars.

4. 4. A method for preparing a baked product according to any one of claims 1 to 3, wherein the flour is selected from the group consisting of wheat flour, all-purpose flour, whole wheat flour, rice flour, corn flour, millet flour, soy flour, oat flour, pastry flour, bread flour, gluten-free flour, tapioca flour and self-rising flour.

5. A method for preparing a baked product according to any one of claims 1 to 4, wherein the dough comprises from 40% to 70% by weight of one or more added flours.

6. 6. A method for preparing a baked product according to any one of claims 1 to 5, wherein the lipase is a Thermomyces lanuginosus (formerly known as Humicola lanuginosus) wild-type lipase.

7. 7. The method for preparing a baked product according to any one of claims 1 to 6, wherein the one or more lipases are polypeptides selected from the group consisting of: (a) a polypeptide having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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%, at least 99% or 100% sequence identity to SEQ ID NO:1; (b) a polypeptide derived from SEQ ID NO: 1 by having 1 to 30 modifications, such as substitutions, deletions and / or insertions, at one or more positions, for example 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 modifications, particularly substitutions; (c) a polypeptide derived from the polypeptide of (a), wherein the N-terminus and / or C-terminus is extended by the addition of one or more amino acids; and (d) a fragment of the polypeptide of (a).

8. 8. A method for preparing a baked product according to any one of claims 1 to 7, wherein the one or more lipases are polypeptides having the sequence of SEQ ID NO:

1.

9. 9. The method for preparing a baked product according to any one of claims 1 to 8, wherein the one or more proteases are polypeptides selected from the group consisting of: (a) a polypeptide having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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%, at least 99%, or 100% sequence identity to SEQ ID NO:6; (b) a polypeptide derived from SEQ ID NO: 6 by having 1 to 30 modifications, such as substitutions, deletions and / or insertions, at one or more positions, for example 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 10 or 11 or 12 or 13 or 14 or 15 or 16 or 17 or 18 or 19 or 20 or 21 or 22 or 23 or 24 or 25 or 26 or 27 or 28 or 29 or 30 modifications, particularly substitutions; (c) a polypeptide derived from the polypeptide of (a), wherein the N-terminus and / or C-terminus is extended by the addition of one or more amino acids; and (d) a fragment of the polypeptide of (a).

10. A method for preparing a baked product according to any one of claims 1 to 9, wherein the one or more proteases are polypeptides having the sequence of SEQ ID NO:

6.

11. A method for preparing a baked product according to any one of the preceding claims, wherein the dough comprises from 12% to 35% by weight of one or more fats.

12. 12. The method for preparing a baked product according to any one of claims 1 to 11, wherein the baked product has between 1% and 5% less sugars by weight compared to a baked product prepared without the one or more lipases and / or one or more proteases.

13. 1. Use of one or more lipases and / or one or more proteases in the production of a baked product to reduce 10% to 25% by weight of fat and / or 1% to 5% by weight of sugars used in the preparation of said baked product.

14. 14. The use of claim 13, wherein one or more properties of the baked product are maintained, at least one of said properties being selected from the group consisting of mouthfeel, texture, taste, chewiness, spreadability, color-height-length, volume, moisture, strength, hardness, crispness and shelf life.

15. A baked product obtainable by the method according to any one of claims 1 to 12 or by the use according to claim 13 or 14.

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