TECHNICAL FIELD
[001] The subject matter described herein in general relates to food products, and in particular, relates to enzymatic compositions for preparing reduced gluten food products.
BACKGROUND OF INVENTION
[002] Gluten, a structural protein, commonly found in cereal grain flours such as, wheat, oats, barley, and rye, and other food products such as pasta, malt, semolina, oats, energy bars, etc., is known to impart viscoelasticity and cohesiveness to dough and other food products. However, some individuals cannot consume certain cereal grain flours and gluten containing food products because they are allergic or cannot easily digest gluten. Ingestion of gluten in these individuals who are sensitive to gluten would trigger a chain of immunological reactions ultimately leading to enteropathy and malabsorption syndrome. (Shewry PR., Wheat; J Exp Bot. 2009;60(6): 1537-53). These patients are generally suggested to adhere to a gluten-free diet or very-low gluten diet. However, because of its ubiquitous presence in several cereal grains, and food products, it becomes extremely difficult to restrict or prohibit gluten containing food.
[003] Attempts to make gluten free food products meant a compromise on texture, viscoelastic behavior and cohesiveness to the food products. For example, a dough prepared by mixing gluten free cereal flour does not rise as efficiently as the gluten containing flour. Dough binder alternatives such as, guar gum, xanthan gum and/or modified starch are conventionally used, these alternatives often do not provide the structure, texture, and expansion demanded to be similar to gluten-containing foods, and furthermore, require a sacrifice of taste, texture and/or appearance of the final product as compared to those gluten-containing foods. Although various methods to produce hypoallergic or reduced-gluten wheat flour are known in the art, these methods available are time consuming, cost ineffective, temperature and pH specific involving multimodal steps. SUMMARY OF THE INVENTION

[004] In an aspect of the present invention, there is provided an a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15.
[005] In an aspect of the present invention, there is provided a process for preparing an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, said process comprising steps of: a) obtaining the protease enzyme (EC 3.4.23.18); b) obtaining the xylanase enzyme (EC 3.2.1.8); c) obtaining the alpha amylase enzyme (EC 3.2.1.1); and d) contacting the protease enzyme (EC 3.4.23.18), and the xylanase enzyme (EC 3.2.1.8) with the alpha amylase enzyme (EC 3.2.1.1) to obtain the composition.
[006] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description and appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.
BRIEF DESCRIPTION OF THE DRAWINGS
[007] The following drawings form part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better be understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein. [008] Figure 1 represents the effect of various enzymes at combined concentrations of lOOppm with respect to percentage reduction in gluten in comparison to the control, in accordance with an embodiment of present disclosure.

[009] Figure 2 represents the effect of protease at various w/w ratios towards
reduction of gluten in wheat flour, in accordance with an embodiment of present
disclosure.
[0010] Figure 3 represents effect of xylanase at various w/w ratios towards
reduction of gluten in wheat flour, in accordance with an embodiment of present
disclosure.
[0011] Figure 4 represents effect of alpha amylase at various w/w ratios towards
reduction of gluten in wheat flour, in accordance with an embodiment of present
disclosure.
[0012] Figure 5 represents effect of a combination of protease, xylanase, and
amylase at various w/w/w ratios, in accordance with an embodiment of present
disclosure.
DETAILED DESCRIPTION OF THE INVENTION
[0013] Those skilled in the art will be aware that the present disclosure is subject to variations and modifications other than those specifically described. It is to be understood that the present disclosure includes all such variations and modifications. The disclosure also includes all such steps, features, compositions, and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any or more of such steps or features. Definitions
[0014] For convenience, before further description of the present disclosure, certain terms employed in the specification, and examples are delineated here. These definitions should be read in the light of the remainder of the disclosure and understood as by a person of skill in the art. The terms used herein have the meanings recognized and known to those of skill in the art, however, for convenience and completeness, particular terms and their meanings are set forth below.
[0015] The articles "a", "an" and "the" are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.

[0016] The terms "comprise" and "comprising" are used in the inclusive, open
sense, meaning that additional elements may be included. It is not intended to be
construed as "consists of only".
[0017] Throughout this specification, unless the context requires otherwise the
word "comprise", and variations such as "comprises" and "comprising", will be
understood to imply the inclusion of a stated element or step or group of element or
steps but not the exclusion of any other element or step or group of element or
steps.
[0018] The term "including" is used to mean "including but not limited to".
"Including" and "including but not limited to" are used interchangeably.
[0019] Ratios, concentrations, amounts, and other numerical data may be presented
herein in a range format. It is to be understood that such range format is used
merely for convenience and brevity and should be interpreted flexibly to include
not only the numerical values explicitly recited as the limits of the range, but also
to include all the individual numerical values or sub-ranges encompassed within
that range as if each numerical value and sub-range is explicitly recited. For
example, a temperature ranges of about 25-35 °C should be interpreted to include
not only the explicitly recited limits of about 25 °C to about 35 °Q but also to
include sub-ranges, such as 25-30 °Q 28-35 °Q and so forth, as well as individual
amounts, including fractional amounts, within the specified ranges, such as 25.2 °Q
and 32.5 °Q for example.
[0020] The term "at least one" is used to mean one or more and thus includes
individual components as well as mixtures/combinations.
[0021] The term "composition" and "enzymatic composition" have been used
interchangeably throughout the specification.
[0022] Unless defined otherwise, all technical and scientific terms used herein have
the same meaning as commonly understood by one of ordinary skill in the art to
which this disclosure belongs. Although any methods and materials similar or
equivalent to those described herein can be used in the practice or testing of the
disclosure, the preferred methods, and materials are now described. All
publications mentioned herein are incorporated herein by reference.

[0023] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally-equivalent products, compositions, and methods are clearly within the scope of the disclosure, as described herein. [0024] As mentioned previously, conventionally methods to reduce gluten are based on avoiding or restricting foods that contain gluten. However, given the vast majority of foods that contain gluten, abstaining from such food products is a quite challenging. Also, currently used gluten substitutes are a compromise on texture, taste and/or consistency. Conventionally used methods to make reduced-gluten wheat flour are time consuming, cost ineffective, temperature and pH specific involving multimodal steps. The present invention is directed towards the use of an enzymatic composition to reduce gluten in dough of various cereal grains and food products. Although the description herein presented refers to reduced gluten in wheat dough, the proposed composition may also be used to reduce gluten in other cereal grains such as rye, wheat, barley, and food products, including, but not limited to, pasta, energy bars, noodles, breads and pastries, baked foods such as cakes, cookies, pie crusts, brownies, tortillas, beer and malt beverages. [0025] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15. In another embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15. In another embodiment the enzyme composition comprising a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio

in a range of 3:1:1.33 to 3:1:12. In yet another embodiment the enzyme composition comprising a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:1.0 to 3:1:13. [0026] In an embodiment of the present disclosure, there is provided an enzyme composition as disclosed herein, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:1.30 to 3:1:12.
[0027] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:1.30 to 3:1:12. [0028] In an embodiment of the present disclosure, there is provided an enzyme composition as disclosed herein, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio of 3:1:1.33.
[0029] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio is 3:1:1.33
[0030] In an embodiment of the present disclosure, there is provided an enzyme composition as disclosed herein, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio of 3:1:4.
[0031] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the

protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio is 3:1:4
[0032] In an embodiment of the present disclosure, there is provided an enzyme composition as disclosed herein, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio of 3:1:12.
[0033] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio is 3:1:12.
[0034] In an embodiment of the present disclosure, there is provided an enzyme composition as disclosed herein, wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g. In another embodiment, the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 9,000-11,000 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3800-4500 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 110000-115000 SKB U/g. In yet another embodiment, the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 10,000 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 4000 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 100000 SKB U/g.
[0035] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in

a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g.
[0036] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g.
[0037] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:1.30 to 3:1:12; and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g.
[0038] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio of 3:1:1.33; and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g.
[0039] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase

enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio of 3:1:4; and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKBU/g.
[0040] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio of 3:1:12; and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g.
[0041] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the composition further comprises water. [0042] In an embodiment of the present disclosure, there is provided an an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the composition further comprises water.
[0043] In an embodiment of the present disclosure, there is provided an an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), and d) water wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme

activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g.
[0044] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition. In another embodiment, the protease enzyme (EC 3.4.23.18) has a concentration in a range of 18.75 - 56.25 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in a range of 6.25-18.75 ppm with respect to the composition, and the alpha amylase has a concentration in the range of 25- 75ppm with respect to the composition. In yet another embodiment, the protease enzyme (EC 3.4.23.18) has a concentration in a range of 16.75 - 58.25 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in a range of 6.0-19.0 ppm with respect to the composition, and the alpha amylase has a concentration in the range of 22-78 ppm with respect to the composition.
[0045] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.
[0046] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: : a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of

12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.
[0047] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: : a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:1.30 to 3:1:12, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.
[0048] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: : a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:1.33, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.
[0049] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: : a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:4, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.

[0050] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: : a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:12, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.
[0051] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: : a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.
[0052] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) has a w/w ratio in the range of 4:1 - 18:1, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 80-95 ppm, and the xylanase enzyme (EC 3.2.1.8) has a concentration in a range of 5-15 ppm with respect to the composition.
[0053] In an embodiment of the present disclosure, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase

enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), and d) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15., and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, and the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 80-95 ppm, and the xylanase enzyme (EC 3.2.1.8) has a concentration in a range of 5-15 ppm with respect to the composition.
[0054] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the composition further comprises at least one excipient. In an embodiment, the at least one excipient is food product or a cereal grain flour. In another embodiment, the at least one excipient is wheat flour. In yet another embodiment of the present disclosure, the at least one excipient is a pH modifier.
[0055] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), and d) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15, and wherein the composition comprises at least one excipient. [0056] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), and d) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15, and wherein the composition comprises at least one excipient. [0057] In an embodiment of the present invention, there is provided an enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water, and e) at

least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15.
[0058] In an embodiment of the present disclosure, there is provided an enzymatic composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the composition comprises at least one excipient. [0059] In an embodiment of the present disclosure, there is provided an enzymatic composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water and e) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in arange of 3:1:0.75 to 3:1:15.
[0060] In an embodiment of the present disclosure, there is provided an enzymatic composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water and e) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g. [0061] In an embodiment of the present disclosure, there is provided an enzymatic composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water and e) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has

a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition. [0062] In an embodiment of the present disclosure, there is provided an enzymatic composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water and e) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.
[0063] In an embodiment of the present disclosure, there is provided an enzymatic composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water and e) wheat flour, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:1.33 to 3:1:12, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 10,000-11,000 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3800-4500 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 100000-115000 SKB U/g, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 18.75 - 56.25 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in a range of 6.25-18.75 ppm with respect to the composition, and the alpha amylase has a concentration in the range of 25- 75ppm with respect to the composition.
[0064] In an embodiment of the present invention, there is provided a gluten reduced dough obtained by employing the enzyme composition: a) a protease

enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water, and e) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15. [0065] In an embodiment of the present disclosure, there is provided a gluten reduced dough obtained by employing the enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15.
[0066] In an embodiment of the present disclosure, there is provided a gluten reduced dough obtained by employing the enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition. [0067] In an embodiment of the present disclosure, there is provided a gluten reduced dough obtained by employing the enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water and e) e) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15. [0068] In an embodiment of the present invention, there is provided a baked product obtained by employing the enzyme composition: a) a protease enzyme (EC

3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water, and e) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15.
[0069] In an embodiment of the present disclosure, there is provided a baked product obtained by employing the enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15.
[0070] In an embodiment of the present disclosure, there is provided a baked product obtained by employing the enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.
[0071] In an embodiment of the present disclosure, there is provided a baked product obtained by employing the enzyme composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), c) an alpha amylase enzyme (EC 3.2.1.1), d) water, and e) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15. [0072] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising the steps of: a) obtaining the protease enzyme

(EC 3.4.23.18); b) obtaining the xylanase enzyme (EC 3.2.1.8); c) obtaining the alpha amylase enzyme (EC 3.2.1.1), and d) contacting the protease enzyme (EC 3.4.23.18), and the xylanase enzyme (EC 3.2.1.8) with the alpha amylase enzyme (EC 3.2.1.1) to obtain the composition.
[0073] In an embodiment of the present invention, there is provided a process for preparing a composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15, said process comprising the steps of: a) obtaining the protease enzyme (EC 3.4.23.18); b) obtaining the xylanase enzyme (EC 3.2.1.8); c) obtaining the alpha amylase enzyme (EC 3.2.1.1), and d) contacting the protease enzyme (EC 3.4.23.18), and the xylanase enzyme (EC 3.2.1.8) with the alpha amylase enzyme (EC 3.2.1.1) to obtain the composition.
[0074] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, said process comprising the steps of: a) obtaining the protease enzyme (EC 3.4.23.18); b) obtaining the xylanase enzyme (EC 3.2.1.8); c) obtaining the alpha amylase enzyme (EC 3.2.1.1), and d) contacting the protease enzyme (EC 3.4.23.18), and the xylanase enzyme (EC 3.2.1.8) with the alpha amylase enzyme (EC 3.2.1.1) to obtain the composition.
[0075] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme

activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g, said process comprising the steps of: a) obtaining the protease enzyme (EC 3.4.23.18); b) obtaining the xylanase enzyme (EC 3.2.1.8); c) obtaining the alpha amylase enzyme (EC 3.2.1.1), and d) contacting the protease enzyme (EC 3.4.23.18), and the xylanase enzyme (EC 3.2.1.8) with the alpha amylase enzyme (EC 3.2.1.1) to obtain the composition.
[0076] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition, said process comprising the steps of: a) obtaining the protease enzyme (EC 3.4.23.18); b) obtaining the xylanase enzyme (EC 3.2.1.8); c) obtaining the alpha amylase enzyme (EC 3.2.1.1), and d) contacting the protease enzyme (EC 3.4.23.18), and the xylanase enzyme (EC 3.2.1.8) with the alpha amylase enzyme (EC 3.2.1.1) to obtain the composition.
[0077] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) a protease enzyme (EC 3.4.23.18); b) a xylanase enzyme (EC 3.2.1.8), and c) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase

enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition, said process comprising the steps of: a) obtaining the protease enzyme (EC 3.4.23.18); b) obtaining the xylanase enzyme (EC 3.2.1.8); c) obtaining the alpha amylase enzyme (EC 3.2.1.1), and d) contacting the protease enzyme (EC 3.4.23.18), and the xylanase enzyme (EC 3.2.1.8) with the alpha amylase enzyme (EC 3.2.1.1) to obtain the composition.
[0078] In an embodiment of the present invention, there is provided a process for preparing reducing gluten content, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), and ii) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15; b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0079] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), and hi) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough. In another embodiment of the present disclosure, incubating the slurry at a temperature in a range of 39°C to 46°C for a time period in a range of 22-26 hours, to obtain a gluten-reduced dough.
[0080] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a

xylanase enzyme (EC 3.2.1.8), and iii) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°Cto 47°Cfor a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0081] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), iii) an alpha amylase enzyme (EC 3.2.1.1), and iv) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough. [0082] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), iii) an alpha amylase enzyme (EC 3.2.1.1), and iv) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry; and c) incubating the

slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0083] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0084] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0085] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), and hi) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75

to 3:1:15; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough. In another embodiment of the present disclosure, the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3.5 to 1:4.5. [0086] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), and iv) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and c) incubating the slurry at a temperature in a range of 38°Cto 47°Cfor a time period in a range of 20-28 hours, to obtain a gluten-reduced dough. In another embodiment of the present disclosure, the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3.5 to 1:4.5.
[0087] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising: i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), and hi) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and c) incubating

the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0088] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1) and iv) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) has a w/w ratio in the range of 4:1 - 18:1; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0089] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising: i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), and iv) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough. [0090] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising: i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to

the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0091] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising: i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and c) incubating the slurry at a temperature in a range of 38°Cto 47°Cfor a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0092] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), and iv) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and wherein the flour is selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and

combinations thereof, and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough. In another embodiment of the present disclosure, the flour is wheat flour. [0093] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), and iv) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and wherein the flour is selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and combinations thereof, and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0094] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), and iv) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and wherein the flour is selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and combinations thereof, and c) incubating the slurry at a temperature in a range of

38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0095] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), and iv) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and wherein the flour is selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and combinations thereof, and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0096] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and wherein the flour is selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and combinations thereof, and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.

[0097] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5; and wherein the flour is selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and combinations thereof, and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough.
[0098] In an embodiment of the present disclosure, there is provided a process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry, wherein the dough comprises a flour and water having a w/v ratio of the flour to water is 1:4, and wherein the flour is wheat flour; and c) incubating the slurry at a

temperature in a range of 40°C to 45 °C for a time period in a range of 22-24 hours, to obtain a gluten-reduced dough.
[0099] In an embodiment of the present disclosure, there is provided a process for
reducing gluten content in a dough, said process comprising the steps of: a)
obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a
xylanase enzyme (EC 3.2.1.8), and hi) an alpha amylase enzyme (EC 3.2.1.1), and
iv) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC
3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of
3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry;
and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time
period in a range of 20-28 hours, to obtain a gluten-reduced dough; and wherein the
gluten content is reduced in a range of 20-28% with respect to the gluten content in
the dough. In another embodiment of the present disclosure, the gluten content is
reduced in a range of 21.5-27.5% with respect to the gluten content in the dough.
[00100] In an embodiment of the present disclosure, there is provided a
process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough; and wherein the gluten content is reduced in a range of 18-28% with respect to the gluten content in the dough.
[00101] In an embodiment of the present disclosure, there is provided a
process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), and hi) an alpha amylase enzyme (EC 3.2.1.1), wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75

to 3:1:15, wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°Cto 47°Cfor a time period in a range of 20-28 hours, to obtain a gluten-reduced dough; and wherein the gluten content is reduced in a range of 18-28% with respect to the gluten content in the dough.
[00102] In an embodiment of the present disclosure, there is provided a
process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) axylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), and iv) at least one excipient, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry, and wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5, and wherein the flour is selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and combinations thereof; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough; and wherein the gluten content is reduced in a range of 18-28% with respect to the gluten content in the dough.
[00103] In an embodiment of the present disclosure, there is provided a
process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry, and wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5, and wherein the flour is

selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and combinations thereof; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough; and wherein the gluten content is reduced in a range of 18-28% with respect to the gluten content in the dough.
[00104] In an embodiment of the present disclosure, there is provided a
process for reducing gluten content in a dough, said process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry, and wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5, and wherein the flour is selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and combinations thereof; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain a gluten-reduced dough; and wherein the gluten content is reduced in a range of 18-28% with respect to the gluten content in the dough.
[00105] In an embodiment of the present disclosure, there is provided a
gluten-reduced dough obtained by a process comprising the steps of: a) obtaining a composition comprising i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°C to

47°C for a time period in a range of 20-28 hours, to obtain the gluten-reduced dough.
[00106] In an embodiment of the present disclosure, there is provided a
gluten-reduced dough obtained by a process comprising the steps of: a) obtaining a composition comprising: i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) water, and v) at least one excipient wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15, and wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition; b) contacting the composition with a dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain the gluten-reduced dough.
[00107] In an embodiment of the present disclosure, there is provided a
gluten-reduced dough obtained by a process comprising the steps of: a) obtaining a
composition comprising: i) a protease enzyme (EC 3.4.23.18); ii) a xylanase
enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), and iv) at least
one excipient wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme
(EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range
of 3:1:0.75 to 3:1:15; b) contacting the composition with a dough to obtain a slurry;
and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time
period in a range of 20-28 hours, to obtain the gluten-reduced dough.
[00108] In an embodiment of the present disclosure, there is provided a
gluten-reduced dough obtained by a process comprising the steps of: a) obtaining a composition comprising: i) a protease enzyme (EC 3.4.23.18); ii) a xylanase enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) at least one excipient, and v) water, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15; b) contacting the composition with a

dough to obtain a slurry; and c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours, to obtain the gluten-reduced dough.
[00109] In an embodiment of the present disclosure, there is provided a
gluten-reduced dough obtained by a process comprising the steps of: a) obtaining a
composition comprising: i) a protease enzyme (EC 3.4.23.18); ii) a xylanase
enzyme (EC 3.2.1.8), hi) an alpha amylase enzyme (EC 3.2.1.1), iv) at least one
excipient, and v) water, wherein the protease enzyme (EC 3.4.23.18) to the
xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a
weight ratio in a range of 3:1:1.33 to 3:1:12; b) contacting the composition with a
dough to obtain a slurry, wherein the dough comprises wheat flour and water
having a w/w ratio of the wheat flour to water of 1:4; and c) incubating the slurry at
a temperature in a range of 38°Cto 47°C for a time period in a range of 20-28 hours,
to obtain the gluten-reduced dough, wherein the gluten content is reduced in a
range of 20.6%-23.43% with respect to the gluten content in the dough.
[00110] In an embodiment of the present disclosure, the enzymatic
composition of the present disclosure is capable of reducing gluten content in a wide variety of cereal grain flours such as, wheat flour, barley flour, rye flour, spelt flour, triticale, and the like; and food products, such as pasta, energy bars, semolina, and baked products. EXAMPLES
[00111] The disclosure will now be illustrated with working examples,
which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply.

[00112] As mentioned previously, conventionally methods to reduce gluten
are based on avoiding or restricting foods that contain gluten. However, given the vast majority of foods that contain gluten, abstaining from such food products is a quite challenging. Also, currently used gluten substitutes are a compromise on texture, taste and/or consistency. Conventionally used methods to make reduced-gluten wheat flour are time consuming, cost ineffective, temperature and pH specific involving multimodal steps. The present invention is directed towards the use of an enzymatic composition to produce reduced gluten wheat dough. Example 1
Wheat sour dough fermentation
[00113] For this purpose, 1:4 ratio of wheat flour (Aashirvaad MP, ITC) was
added to 4 parts of drinking water (Bisleri). A dose of lOOppm of protease, xylanase and alpha amylase from Advanced Enzyme Technology Ltd. were added to the above slurry. The mixture was placed for incubation for 24 hours at 42°C. Example 2
Preparation of samples and reagents towards quantification by S-ELISA
[00114] The sample preparation protocol was followed as per
manufacturer's instructions (R-Biopharm). Briefly, 0.25g or 250ul of homogenized test sample was weighed and 2.5 ml of cocktail solution (R-Biopharm) was added and mixed well. The same was incubated for 40 minutes at 50°C. The sample is let to cool down and mixed with 7.5 ml 80 % ethanol. The vial is closed and shaken for 1 h up-side down at room temperature, centrifuged for 10 minutes at 2500g at room temperature. Supernatant was collected in a screw capped vial. lOOul of the same immediately used for assay. For preparation of reagents, all reagents brought to room temperature before use. The sample diluent was diluted with 4 times distilled water. The antibody enzyme conjugate was diluted 1:11 with 10 times distilled water. The washing buffer was diluted with 9 times distilled water. Example 3
Quantification of gluten by S-ELISA
[00115] 100 ul test samples and standard solutions (Oppb, 5ppb, lOppb,
20ppb, 40ppb, and 80ppb gliadin solution) added to separate wells in duplicates.

The test samples and the standard solutions were incubated for 30 minutes at room temperature. The liquid poured out of the wells and the wells were further washed with 250 ul diluted washing buffer. The washing step was repeated for two more times. Following the above, 100 ul of the diluted enzyme conjugate added to each well and incubated for 30 minutes at room temperature. On completion, the liquid was poured out of the wells and vigorously (three times in a row) washed as mentioned above. Further, 50 ul of substrate and 50 ul of chromogen was added to each well. The plate was gently shaken manually to mix the reagents, and the plate was further incubated for 30 min at room temperature in dark. On completion of the above, 100 ul of the stop reagent to each well. The plate was again shaken manually, and the absorbance was measured at 450 nm and read within 30 minutes. The gliadin concentration in ug/kg (ppb) is read from calibration curve and further multiplied by dilution factor. This result is then multiplied by 2 to obtain the gluten concentration (gliadin usually represents 50 % of the proteins present in gluten). Example 4
Screening of bioactives towards reduction in gluten in wheat
[00116] Five enzymes PDZYME PT 100, PDZYME FA 400, protease,
xylanase, and alpha amylase were screened at lOOppm concentrations each. The first two enzymes obtained from P D Navkar (Bangalore), the next three enzymes obtained from Advanced Enzymes (Mumbai). The enzymes were screened as per examples 1-3. The results are presented in Figure 1. From figure 1, it can be observed that, from among the 4 enzymes that were screened for gluten reduction in wheat flour, the combination of protease and xylanase in a ratio of 75:25, and amylase, both at a combined concentration of lOOppm were found to exhibit maximum reduction in gluten, by about 14.52% and 12.25% respectively, in comparison to the control Example 5 Dose curve of finalized enzymes
[00117] Protease, xylanase, and alpha amylase were chosen for their higher
activity in reducing gluten in wheat flour; and a dose curve study was performed for the same from 18.75, 37.75, 40, 56.25, 81.25, 93.75, 100 and 200 ppm of protease was used to determine the effect of protease on gluten reduction in wheat

flour. These enzymes were screened for dose response as per Example 1-3. The results are presented in Figure 2. Quantitative analysis showed that the percentage reduction in gluten was found to be dose dependent when a concentration of 56.25 ppm of protease, as observed in Figure 2. Maximum reduction was found to be for the concentrations lOOppm and 200ppm, respectively. While lOOppm of protease reduced gluten content in wheat flour by 13.65%, 200 ppm of protease reduced gluten content in wheat flour by 13.71%, in comparison to the control. Since the difference in percentage reduction of gluten was only marginal between the concentrations lOOppm and 200 ppm; a concentration of lOOppm was selected for further experimentation.
[00118] Similar are the findings with enzyme xylanase as can be observed
from Figure 3. Concentrations ranging from 6.25, 12.5, 18.75, 25, 50, 100 and 200
ppm of xylanase was used to determine the effect of xylanase on gluten reduction
in wheat flour. These enzymes were screened for dose response as per Example 1 -
3. Quantitative analysis of figure 3 reveals that the percentage reduction was dose
dependent. Maximum reduction was found to be for the concentrations lOOppm
and 200ppm, respectively. While lOOppm of xylanase reduced gluten content in
wheat flour by 8.32%, 200 ppm of xylanase reduced gluten content in wheat flour
by 9.64%, in comparison to the control. Since the difference in percentage
reduction of gluten was only marginal between the concentrations lOOppm and 200
ppm; a concentration of lOOppm was selected for further experimentation.
[00119] Figure 4 demonstrates the effect of alpha amylase at various w/w
ratios on gluten reduction in wheat flour. Concentrations ranging from 25, 50, 75, 100 and 200 ppm of alpha amylase was used to determine its effect of on gluten reduction in wheat flour. These enzymes were screened for dose response as per Example 1-3. Quantitative analysis of figure 4 reveals that the percentage reduction was dose dependent. Maximum reduction was found to be for the concentrations lOOppm and 200ppm, respectively. While lOOppm of alpha amylase reduced gluten content in wheat flour by 10.07%, 200 ppm of xylanase reduced gluten content in wheat flour by 10.41%, in comparison to the control. Since the difference in percentage reduction of gluten was only marginal between the

concentrations lOOppm and 200 ppm; a concentration of lOOppm was selected for
further experimentation.
Example 6
Evaluation of combination of protease, xylanase, and alpha amylase at various
w/w/w ratios on reduction of gluten in wheat flour
[00120] Based on the results obtained, effect of combinations of protease,
xylanase, and alpha amylase, in various w/w rations was tested, wherein the
maximum individual concentration of the actives was lOOppm for either of
enzymes. The ratios and concentrations tested are mentioned below in Table 1:
[00121] For the various w/w ratios tested, unexpectedly and surprisingly, the
combinatorial effect was more than the effect of either ingredient alone, the combinatorial effect being synergistic for the ratios. From a combined reading of Table 1, Figure 2, Figure 3, Figure 4, and Figure 5, it is observed that when the actives were used in combination at different ratios, the three actives in combination exhibit a synergistic effect, particularly at ratios of 56.25:18.75:25, 37.5:12.5:50, and 6.25:18.75:75. The inhibition was found to be maximum for the ratios of 6.25:18.75:75 and 56.25:18.75:25. It was found to be 26.69% and 22.06% respectively.
Advantages of the present disclosure
[00122] The present disclosure reveals an enzymatic composition to reduce
gluten in a wide variety of cereal flours such as wheat flour, barley flour, rye flour, spelt flour, and other food products containing gluten. The reduced gluten wheat

flour produced in the present invention may be used as a 1-for-l substitute in any conventional wheat flour recipes, thus eliminating the necessity of using exclusively gluten-free recipes for gluten sensitive individuals. The flour produced thus can be used to make batter, roti's, cookies, cakes, muffins, sauces, and other baked products.

I/We Claim;
1. An enzyme composition comprising:
a) a proteas e enzyme (EC3.4.23.18);
b) a xylanase enzyme (EC 3.2.1.8); and
c) an alpha amylase enzyme (EC 3.2.1.1)
wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:2:15.
2. The composition as claimed in claim 1, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:0.75 to 3:1:15.
3. The composition as claimed in claim 1, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio in a range of 3:1:1.30 to 3:1:12.
4. The composition as claimed in claim 1, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio of 3:1:1.33.
5. The composition as claimed in claim 1, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio of 3:1:4.
6. The composition as claimed in claim 1, wherein the protease enzyme (EC 3.4.23.18) to the xylanase enzyme (EC 3.2.1.8) to the alpha amylase enzyme (EC 3.2.1.1) has a weight ratio has a weight ratio of 3:1:12.
7. The composition as claimed in claim 1, wherein the protease enzyme (EC 3.4.23.18) has an enzyme activity in a range of 8500-11500 PCu/g, the xylanase enzyme (EC 3.2.1.8) has an enzyme activity in a range of 3400-4600 XU/g, and the alpha amylase (EC 3.2.1.1) has an enzyme activity in a range of 85000-115000 SKB U/g.
8. The composition as claimed in claim 1, wherein the composition further comprises water.

9. The composition as claimed in any one of the claims 1-8, wherein the protease enzyme (EC 3.4.23.18) has a concentration in a range of 12-60 ppm, the xylanase enzyme (EC 3.2.1.8) has a concentration in the range of 5-20 ppm, and the alpha amylase (EC 3.2.1.1) has a concentration in the range of 15-85 ppm with respect to the composition.
10. The composition as claimed in any one of the claims 1-9, wherein the composition further comprises at least one excipient.
11. A process for preparing the composition as claimed in claim 1, said method comprising:

a) obtaining the protease enzyme (EC 3.4.23.18);
b) obtaining the xylanase enzyme (EC 3.2.1.8); and
c) obtaining the alpha amylase enzyme (EC 3.2.1.1)
d) contacting the protease enzyme (EC 3.4.23.18), and the xylanase enzyme (EC 3.2.1.8) with the alpha amylase enzyme (EC 3.2.1.1) to obtain the composition.
12. A process for reducing gluten content in a dough, said process comprising:
a) obtaining the composition as claimed in any one of the claims 7-10;
b) contacting the composition with a dough to obtain a slurry; and
c) incubating the slurry at a temperature in a range of 38°C to 47°C for a time period in a range of 20-28 hours,
to obtain a gluten-reduced dough.
13. The process as claimed in claim 12, wherein the dough comprises a flour and water having a w/v ratio of the flour to water in a range of 1:3 to 1:5.
14. The process as claimed in claim 13, wherein the flour is selected from a group consisting of wheat flour, barley flour, rye flour, spelt flour, triticale, and combinations thereof.
15. The process as claimed in claim 12, wherein the process leads to reduction of gluten content in a range of 20-28% with respect to the gluten content in the dough.
16. A gluten-reduced dough obtained using the process as claimed in claim 12.

17. A gluten-reduced dough obtained by employing the composition as claimed in claim 1-10.
18. A baked product obtained by employing the composition as claimed in claim 1-10