BACKGROUND OF THE INVENTION

Field of the Invention

The present invention is broadly concerned with an enzyme composition for use in lieu of diacetyl tartaric acid esters of mono- and diglycerides ("DATEM") in baked products, such as bread.

Description of the Prior Art

Dough conditioners are commonly used in baked products to improve the properties and handling of the dough, along with the properties of the final baked products. These properties include volume, texture, flavor, freshness, and machinability. Dough conditioners are often typical ly chemical additives, such as emulsifiers (e.g., DATEM). Consumers are becoming more and more resistant to chemical additives in food. However, it has been difficult to achieve the desired properties without these additives. There is a need for further non-chemical options that can achieve properties similar to that of the currently available chemical additives.

SUMMARY OF THE INVENTION

The present invention is concerned with an enzyme mixture useful for preparing baked products. The mixture comprises a first xylanase, a first lipase, and an enzyme selected from the group consisting of: a second xylanase different from the first xylanase; a second lipase different from the first lipase; and both the second xylanase and second lipase. The mixture is also substantially free of one or more of the following enzymes: cellulases, glucose oxidases, and trans-glutaminases.

The invention further provides a dough including flour and other dough-making ingredients, the improvement being that the dough further comprises:

a first xylanase;

a first l ipase; and

an enzyme selected from the group consisting of a second xylanase different from the first xylanase, a second lipase different from the first lipase, and both the second xylanase and second l ipase, wherein the dough is substantially free of one or more of the following enzymes: cellulases, glucose oxidases, and trans-glutaminases.

The invention is further directed towards a method of form ing a bakery product. The method comprises providing a dough comprising:

flour;

a first xylanase;

a first l ipase; and

an enzyme selected from the group consisting of a second xylanase different from the first xylanase, a second lipase different from the first lipase, and both the second xylanase and second lipase. The dough is also substantially free of one or more of the fol lowing enzymes: cellulases, glucose oxidases, and trans-glutam inases. The dough is then baked for a time and temperature sufficient to yield the bakery product.

In a further embodiment, the invention provides a dough including flour and other dough-making ingredients where the improvement is that the dough further comprises a first xylanase, a first lipase, and lecithin.

Additionally the invention provides a method of forming a bakery product where the method comprises provid ing a dough. The dough comprises flour, a first xylanase, a first lipase, and lecithin. The dough is baked for a time and temperature sufficient to yield the bakery product.

In yet a further embodiment, an enzyme mixture is provided that comprises a first xylanase, a first lipase, and lecithin.

DETA ILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

In more detail, the present invention is concerned with novel enzyme mixtures that can be used to make dough for forming yeast-raised and other bakery products. These products include those selected from the group consisting of breads, pretzels, English muffins, buns, rolls, tortillas (both corn and flour), pizza dough, bagels, and crumpets.

Inventive Enzyme Mixture

The inventive enzyme m ixture broadly comprises a first xylanase and a first lipase as well as a second xylanase different from the first xylanase and/or a second lipase different from the first lipase and/or lecithin. In preferred embodiments, the enzyme m ixture comprises both the second xylanase and the second lipase and/or lecithin. In a further preferred embodiment, it also comprises a fungal amylase and/or lecithin. In a further embodiment, the enzyme mixture consists essentially of, or even consists of, the foregoing variations, optional ly with a carrier.

In more detail, the first xylanase and second xylanase (when included) are preferably bacterial or fungal xylanases that act on hemicelluloses present in dough during the baking process. The preferred xylanases will have a minimum activity of from about 300 XylH/g to about 2,500 XylH/g and preferably from about 500 XylH/g to about 2, 100 XylH/g. As referred to herein, " 1 XylH" corresponds to an enzyme that releases from xylan a quantity of reducing groups equivalent to 1 μΜοΙ xylose per minute at 30C under standard conditions using the hydrazid method.

Even more preferably, one xylanase wi ll have a m inimum activity of from about 300

XylH/g to about 800 XylH/g and preferably from about 500 XylH/g to about 700 XylH/g, and one xylanase wi ll have a minimum activity of from about 1 ,200 XylH/g to about 2,500

XylH/g and preferably from about 1 ,500 XylH/g to about 2, 100 XylH/g. If only one type of xylanase is present, then that xylanase wi ll have an activity in one of the foregoing ranges.

In a preferred embodiment, the first and second xylanases are individual ly derived from an organism selected from the group consisting of Bacillus and Aspergillus. Even more preferably, the first and second xylanases are derived from an organism selected from the group consisting of Bacillus licheniformis, Bacillus subtilis, and Aspergillus niger. It will be appreciated that the first and second xylanases could be derived from the same or different organism genus, or even the same or different species, provided they are different xylanases.

The total quantity of xylanase(s) in the inventive mixture wi ll be from about 0.2% to about 50% by weight, preferably from about 0.3% to about 30% by weight, and more preferably from about 0.4%o to about 20% by weight, based upon the total weight of the enzyme m ixture taken as 1 00% by weight. In embodiments where at least two xylanases are present, one or more xylanases are present in the inventive enzyme mixture at levels of from about 0.2% to about 3% by weight each, preferably from about 0.5% to about 2.5% by weight each, and more preferably from about 0.7% to about 2% by weight each. One or more other xylanases are included at levels of from about 0.2% to about 20.0% by weight each, preferably from about 0.3% to about 1 0% by weight each, and more preferably from about 0.4% to about 5% by weight each, based upon the total weight of the enzyme mixture taken as 1 00% by weight.

The first l ipase and second lipase (when included) are preferably Humicola (also known as Thermomyces) l ipases that break down fat and lipids during the baking process and convert them to glycerol and free fatty acids. The preferred lipases wi ll have a minimum activity of from about 3 LU/g to about 1 00 KLU/g, and preferably from about 6 KLU/g to about 75 KLU/g. The activity of this enzyme is measured in KLU/g. As used herein, the LU method is based on the speed at which the enzyme hydrolyses tributyrin at pH 7.0. The butyric acid formed is titrated with sodium hydroxide, and the consumption of the latter recorded as a function of time. 1 LU is the amount of enzyme, which releases 1 pmol butyric acid per minute under the given standard conditions. 1 KLU = 1 ,000 LU.

Even more preferably, one lipase will have a minimum activity of from about 3 KLU/g to about 14 KLU/g and preferably from about 5 KLU/g to about 1 0 KLU/g, and one lipase will have a minimum activity of from about 25 KLU/g to about 100 KLU/g and preferably from about 35 KLU/g to about 75 KLU/g. I f only one type of lipase is present, then that lipase will have an activity in one of the foregoing ranges.

In a preferred embodiment, the first and second lipases are individually derived from an organism selected from the group consisting of Humicola and Fusarium. Even more preferably, the first and second lipases are derived from an organism selected from the group consisting of Humicola lanuginosa and Fusarium oxysporum. It wi ll be appreciated that the first and second lipases could be derived from the same or different organism genus, or even the same or different species, provided they are different lipases.

The total quantity of l ipase(s) in the inventive mixture will be from about 0.01 % to about 50% by weight, preferably from about 0.02% to about 30% by weight, and more preferably from about 0.03% to about 20% by weight, based upon the total weight of the enzyme mixture taken as 100% by weight. In embodiments where at least two lipases are present, one or more lipases are included in the inventive enzyme mixture at levels of from about 0.0 1 % to about 3% by weight each, preferably from about 0.02% to about 2% by weight each, and more preferably from about 0.03% to about 1 % by weight each. One or more other lipases are included at levels of from about 0.3% to about 20% by weight each, preferably from about 0.4% to about 1 0% by weight each, and more preferably from about 0.5% to about 5% by weight each, based upon the total weight of the enzyme mixture taken as 1 00% by weight.

In some embodiments, the enzyme mixture further comprises a fungal amylase (preferably a fungal alpha-amylase) with any of the lipase and xylanase combinations described above. The fungal amylase will act on starch, and the preferred fungal amylase wi ll have a minimum activity of from about 500 FAU-F/g to about 3,000 FAU-F/g and preferably from about 1 ,500 FAU-F/g to about 2,600 FAU-F/g. As used herein, FAU(F) Fungal alpha-amylase Units (Fungamyl) are measured relative to an enzyme standard of a declared strength. The assay substrate is 4,6-ethylidene(G7)-p-nitrophenyl(G l )-alpha, D- maltoheptaoside (ethylidene-G7-PNP). Reaction conditions:

Temperature 37° C; pH 7. 15 ; Wavelength 405 nm; Reaction time 5 min; and Measuring time 2 min.

In a preferred embodiment, the fungal amylase is derived from Aspergillus, and more preferably, from Aspergillus oryzae. (n embodiments where a fungal amylase is included, the quantity of the fungal amylase in the inventive enzyme mixture will be from about 0.2% to about 20% by weight, preferably from about 0.3% to about 10% by weight, and more preferably from about 0.4% to about 5% by weight, based upon the total weight of the enzyme mixture taken as 1 00% by weight.

As noted previously, lecithin can be included in the enzyme mixture, in any of the various enzyme combinations described previously. When lecithin is included, it is present at levels of from about I to about 50% by weight, more preferably from about 5 to about 45% by weight, and even more preferably from about 1 0 to about 40% by weight, based upon the total weight of enzyme m ixture taken as 100% by weight.

It is preferred that the inventive enzyme m ixture be substantial ly free (i.e., comprises less than about 1 % by weight, preferably less than about 0.5% by weight, and preferably about 0% by weight) of one or more of the following enzymes: cellulases, glucose oxidases, and trans-glutaminases. More preferably, the enzyme m ixture is substantially free of at least two of these enzymes, and even more preferably the enzyme mixture is substantially free of al l three of these enzymes.

Preferred weight ratios of the various enzymes are shown in Table A. Table A - Weight Ratios ^A

^A Weight ratios also applicable to inventive dough.

^B In embodiments where a fungal amylase is present.

Final ly, it wi ll be appreciated that the enzyme mixture can be provided with purely the above enzymes and/or lecithin, or il can be dispersed in a carrier powder (e.g., flour, calcium sulfate, starch, silicon dioxide) alone or with other optional ingredients that might be useful during the baking process. Examples of suitable optional ingredients include those selected from the group consisting of sodium ch loride, water, sunflower oil, wheat flour, starch, and dextrin.

Inventive Dough Ingredients

The inventive enzyme mixture will be included in the dough at levels such that the dough will include each enzyme at the activity levels set forth in Table B. Lecithin levels are also set forth in fable B.

Table B

( Xyl H/kg of Hour) 1000

Fungal Amylase* * from about 0. 1 to about 1 000 from about 10 to about 300 from about 30 to about 100 (FA U-F/kg of flour)

Lecithin** (% based from about 0.01 to about 10 from about 0. 1 to about 5 from about 0.2 to about 2 on flour)

* In embodiments where more than one lipase and/or more than one xylanase are present. * * In embodiments where a fungal amylase and/or lecithin are present.

Typical baked product ingredients and their preferred ranges are set forth in Table C, although it wil l be appreciated that these are exemplary and can be adjusted depending upon the formulation of the end user.

Table C

*Percentage or ppm based upon the weight of flour.

** In embodiments where still included.

In addition to the ingredients from Table C, the dough will include a source of starch, which will typicall be a flour. Preferred such sources of starch include those selected from the group consisting of wheat flour, rye flour, oat flour, barley flour, triticale flour, rice flour, tapioca starch, corn starch, wheat starch, rice starch, potato starch, corn flour, and potato flour. The source of starch will typically be included to provide levels of from about 50% to about 95% by weight starch, and preferably from about 65% to about 85% by weight starch, based upon the tota l weight of the flour taken as 100% by weight. When flour is the source of starch, this wil l typical ly result in flour levels of from about 40% to about 70% by weight flour, and preferably from about 50% to about 60% by weight flour, based upon the total weight of the dough taken as 100% by weight.

The yeast used can be any yeast conventionally used in yeast-raised bakery products, with cream and compressed yeast being preferred. Preferred mold inhibitors include those selected from the group consisting of calcium and/or sodium propionate, potassium sorbate, vinegar, raisin juice concentrate, and mixtures thereof. The preferred oil or fat is selected from the group consisting of soy oil, partially hydrogenated soy oil, lard, palm oil, corn oil, cottonseed oil, canola oil, and mixtures thereof.

Suitable flour improvers include ascorbic acid. While any conventional emulsi fier can be util ized, preferred emulsifiers include polyoxyethylene sorbitan monostcarate (typical ly referred to as Polysorbate 60) and monoglycerides, such as powdered and hydrated monoglycerides, citrated monoglycerides, and succinylated monoglycerides.

It is preferred that the dough be substantial ly free (i.e., comprises less than about 0.5% by weight, preferably less than about 0.1 % by weight, and preferably about 0% by weight) of one or more of the following enzymes: cellulases, glucose oxidases, and transglutam inases. More preferably, the dough is substantially free of at least two of these enzymes, and even more preferably the dough is substantially free of al l three of these enzymes.

Furthermore, it wi ll be appreciated that the inventive enzyme mixture allows for

DATEM and other ingredients that do not allow for a "clean" label (e.g., dough conditioners such as sodium stearoyi lactylate and ethoxylated monoglyceride; flour improvers such as potassium bromate, potassium iodate, azodicarboamide, and calcium peroxide) to be substantially reduced, and even eliminated, from the dough used in the baking process. In instances where DA TEM is substantial ly reduced, the dough will comprise less than about

0.5% by weight DATEM, preferably less than about 0.3% by weight DATEM, and more preferably from about 0.01 % to about 0.2% by weight DATEM, based upon the total weight of the flour taken as 1 00% by weight. In instances where DATEM is elim inated, the dough will be substantial ly free of DATEM (i.e., comprises less than about 0.01 % by weight, preferably less than about 0.005% by weight, and preferably about 0% by weight DATEM).

It is preferred that the dough is also substantially free of (i.e., comprises less than about

0.005% by weight, preferably less than about 0.001 % by weight, and preferably about 0% by weight) one or more (and preferably all) of the following: sodium stearoyl lactylate, ethoxylated monoglyceride, potassium bromate, potassium iodate, azodicarbonamide, and calcium peroxide.

Method of Making Baked Products

In forming the dough according to the invention, the above ingredients can be simply m ixed together in one stage using the "no-time dough process," or they can be subjected to the "sponge and dough process." In the "no-time dough process," all ingredients are added to a mixing bowl at the same lime and m ixed for a time period from about 5 to about 1 5 minutes to form the mixed dough.

In the "sponge and dough" process, part of the flour (e.g., 55-75% by weight of the total flour) is mixed with water, yeast, and preferably the dough strengthener (if utilized) and allowed to ferment for a time period of from about 3 hours to about 4 hours. This forms the "sponge." After this time period, the remaining ingredients are mixed with the sponge for a time period of from about 2 minutes to about 10 minutes to form the mixed dough. The mixed dough is preferably allowed to rest for a time period of from about 5 minutes to about 1 5 m inutes before be ing formed into the desired size pieces and placed in the baking pans. The dough is then preferably allowed to proof at a temperature of from about 40C to about 50C at a relative humidity of from about 65% to about 95% for a time period of from about 50 minutes to about 70 minutes. After proofing, the product can then be baked using the times and temperatures necessary for the type of product being made (e.g., from about 1 90C to about 220C for about 20 minutes to about 30 minutes).

The invention results in a number of advantages in the final baked product, in addition to those discussed previously. For example, the invention allows the DATEM in the product to be substantially reduced, or even eliminated, at the levels noted previously with respect to the dough. Furthermore, even though DATEM is being reduced and/or avoided, similar volumes, textures, tastes, and cell structures are achieved. For example, the inventive method will retain at least about 80%, preferably at least about 85%, and more preferably at least about 90% of the volume that would be achieved making the same bread with only 50%o or less of the amount of DATEM that would otherwise be included. Furthermore in embodiments where lecithin is included, inventive method will retain at least about 93%, preferably at least about 96%, more preferably at least about 99%, and even more preferably 100% of the volume that would be achieved making the same bread with only 50% or less of the amount of DATEM that would otherwise be included. Volume is determined as described in Example 1 A.

EXAMPLES

The following examples set forth preferred methods in accordance with the invention. It is to be understood, however, that these examples are provided by way of illustration and nothing therein should be taken as a limitation upon the overall scope of the invention.

EXAMPLE 1 A

Xylanase Testing

In this procedure, the performance of varying quantities of Veron 1 9 1 (a xylanase, obtained from specific cultures of Aspergillus niger, and available from A B Enzymes) was tested in a 100% whole wheat bread formulation and compared against that of DATEM. The formulation and variations tested are shown in Tables 1 and 2.

TABLE 1

100% Whole Wheat Bread Formula

Ingredients % of Flour Weight

Whole Wheat Flour 100

Vital Wheat Gluten 4

Granulated Sugar 1 0

Salt 2

Calcium Propionate 0.25

Ascorbic Acid 0.012

Soy Oil 2

Yeast-Dry 2

Water 70 TABLE 2

Formu la Variations

^A Fungamyl 2500 is a fungyl alpha-amylase derived from Bacillus subtilis and available from Novozymes.

^B Lipopan F Cone is a Fusarium l ipase expressed by submerged fermentation in a genetically modified Aspergillus oryzae organism. Available from Novozymes.

c BXP 25001 is a xylanase derived from Bacillus subtilis, available from DSM (DSM determines xylanase activity differently than the definition being used herein. It's defined on 1 % wheat arabinoxylan at pH 6.0, T= 30°C, and an incubation time of 30 min. The xylose reducing residues are measured with a reducing sugar assay. One NBXU is defined as the amount of enzyme that can produce 0.5 mg of xylose reducing residues in the incubation mixture under the assay conditions.)

The ingredients were added to a FJobart mixer, and mixed unti l optimal development. The mixer bowl was chilled by circulating 20°C chil ling water through the cooling jackets of the mixing bowl . After mixing, the dough was allowed to rest on a wood bench for 10 minutes and then, divided, sheeted, and moulded according to the processing parameters shown in Table 3.

TABLE 3

Processing Parameters

Molding Type Straight

Proof Condition 104F/78% RH

To height- 1 .7 cm above top

Proof Time

of pan

Baking Temp 420F

Baking Time 20 min

Bag Time 65 min

The moulded dough pieces were then placed into loaf pans and proofed to the targeted height for around 55-60 m in . Each prepared dough was divided into two aliquots, one that was "shocked" and one that was not (referred to as "unshocked"). A dough is shocked by placing it (after proofing, wh i le it's still in its pan) on two wooden blocks that are 8.5 inches apart and dropping it (pan and al l) from a height of 3.375 inches (height of the wooden blocks). This al lows the dough's abi lity to recover to be tested. Each dough (both shocked and unshocked) was then baked to form a bread. After baking, the loaves were left on a metal shelf for cooling for 60 minutes and then packed individually in plastic bags.

The volume of each sample was then determined by a benchtop laser-based scanner called a VolScan Profiler (by Stable Micro Systems), which is a non-contact measurement system that measures product volume and provides a rapid, 3-dimensional digitization of products. Vertical step was run at 4 mm, and Rotation Speed was set to Standard ( l rps). The results are shown in Table 4. It was found that the Veron 191 assisted in obtaining some of the volume properties achieved with DATEM, but without including DATEM . Furthermore, when the Veron 1 91 samples were sliced and visually observed, they were noted to have simi lar cell structure and texture to the DATEM samples.

TAB LE 4 EXAMPLE I B

Xylanase Testing

In this procedure, the performance of varying quantities of Veron 568 (a xylanase obtained from speci fic cultures of Bacillus subtilis, and available from AB Enzymes) was tested in a 1 00% whole wheat bread formulation and compared against that of DATEM. The formulation and variations tested are shown in Tables 5 and 6.

TABLE 5

100% Whole Wheat Bread Formula

TABLE 6

Formula Variations

The above formulation was prepared according to the same processing parameters described in Example 1 A. Again, each prepared dough was divided into two aliquots, one that was "shocked" and one that was not (referred to as "unshocked"). Each dough (both shocked and unshocked) was then baked to form a bread.

The volume of each sample was then determined following the procedure described in Example 1 A. The results are shown in Table 7. It was found that the Veron 568 assisted in obtaining some of the volume properties achieved with DATEM, but without including DATEM. Furthermore, when the Veron 568 samples were sliced and visually observed, they were noted to have similar cell structure and texture to the DATEM samples.

TA BLE 7

EXAMPLE 2A

Lipase Testing

In this procedure, the performance of varying quantities of Lipopan 50 BG (a lipase derived from Aspergillus oryzae, and available from Novozymes) was tested in a 100% whole wheat bread formulation. The formulation and variations tested are shown in Tables 8 and 9. It may be noted that from here on out Novamyl Pro (a Novamyl variant from Novozymes®) was used during testing as a standard shelf-life extending system in order to further represent a typical industrial formula.

TA BLE 8

1 00% Whole Wheat Bread Formula

TABLE 9

Formula Variations

The above formulation was prepared according to the same processing parameters descri bed in Example 1 A. Again, each prepared dough was divided into two aliquots, one that was "shocked" and one that was not (referred to as "unshocked"). Each dough (both shocked and unshocked) was then baked to form a bread. The respective volumes of each sample was then determined as described previously and are shown in Table 10. It was found that the Lipopan 50 BG assisted in obtaining some of the volume properties similar to DATEM, but without including DATEM. Furthermore, when the Lipopan 50 BG samples were sliced and visually observed, they were noted to have similar cell structure and texture to the DATEM samples.

TABLE 1 0

EXAMPLE 2B

Lipase Testing

In this procedure, the performance of varying quantities of Lipopan Xtra BG (a l ipase derived from Aspergillus oryzae, and available from Novozymes) was tested in a 100% whole wheat bread formulation and compared against that of DATEM. The formulation and variations tested are shown in Tables 1 1 and 12.

TABLE 11

100% Whole Wheat Bread Formula

TABLE 12

5 Formula Variations

The above formulation was prepared according to the same processing parameters described in Example 1 A . Again, each prepared dough was divided into two aiiquots, one that was "shocked" and one that was not (referred to as "unshocked"). Each dough (both shocked and unshocked) was then baked to form a bread.

5 The respective volumes of each sample were then determined as described previously and are shown in Table 13. It was found that the Lipopan Xtra BG assisted in obtaining some of the volume properties achieved with DATEM, but without including DATEM . Furthermore, when the Lipopan Xtra BG samples were sl iced and visually observed, they were noted to have simi lar cell structure and texture to the DATEM samples.

1 0

TA BLE 1 3

EXAMPLE 3

1 5 Inventive Enzyme Mixture Testing

In this procedure, the performance of a mixture of the two lipases and two xylanases tested in Examples 1 A, I B, 2 A, and 2B along with a fungal amylase was tested in a 100% whole wheat bread formulation and compared against that of DATEM. The formulation and variations tested are shown in Tables 14 and 1 5.

TABLE 14

100% Whole Wheat Bread Formula

TABLE 15

Formula Variations

The above formulation was prepared according to the same processing parameters described in Example 1 A . Again, each prepared dough was divided into two aliquots, one that was "shocked" and one that was not (referred to as "unshocked"). Each dough (both shocked and unshocked) was then baked to form a bread.

The respective volumes of each sample were then determined as described previously and are shown in Table 16. It was found that the inventive enzyme mixture assisted in obtaining some of the volume properties achieved with DATEM, but without including DATEM . Furthermore, when the samples prepared with the inventive enzyme mixture were sliced and visually observed, they were noted to have sim i lar cell structure and texture to the DATEM samples.

TABLE 1 6

EXAMPLE 4A

Lecithin in Combination with Inventive Enzyme Mixture In this procedure, the performance of an enzyme mixture similar to that of Example 3 along with lecithin (specifical ly, sunflower lecithin powder, obtained from Caldic) was tested in a 1 00% whole wheat bread formulation and compared against that of the enzyme mixture without lecithin present. The formulation and variations tested are shown in Tables 1 7 and 1 8.

TABLE 1 7

100% Whole Wheat Bread Formula

TABLE 1 8

Formula Variations

The above formulation was prepared according to the same processing parameters described in Example 1 A. Again, each prepared dough was divided into two aliquots, one that was "shocked" and one that was not (referred to as "unshocked"). Each dough (both shocked and unshocked) was then baked to form a bread.

The respective volumes of each sample were then determined as described previously and are shown in fable 1 9. It was found that using lecithin with the inventive enzyme m ixture assisted in obtaining even further improved volume. Furthermore, when the samples prepared with the inventive enzyme m ixture were sliced and visually observed, they were noted to have desirable cell structure and textures.

TABLE 19

EXAMPLE 4B

Lecithin in Combination with Inventive Enzyme Mixture In this procedure, an enzyme mixture along with lecithin (simi lar to that of Example 4A) was tested in a 100% whole wheat bread formulation and compared against that of DATEM. The formulation and variations tested are shown in Tables 20 and 2 1 . For the final testing, Novamyl Pro was el iminated and Ultra Fresh Premium 250 (shelf-l i fe extending enzyme m ixture; avai lable from Corbion) to test performance in the presence of other enzymes typically used in the baking industry.

TABLE 20

100% Whole Wheat Bread Formula

TABLE 21

Formula Variations

The above formulation was prepared according to the same processing parameters described in Example 1 A . Again, each prepared dough was divided into two aliquots, one that was "shocked" and one that was not (referred to as "unshocked"). Each dough (both shocked and unshocked) was then baked to form a bread.

The respective volumes of each sample were then determined as described previously and are shown in Table 22. It was found that using lecithin with the inventive enzyme mixture assisted in obtaining volume properties comparable to that of DATE . Furthermore, when the samples prepared with the inventive enzyme mixture and lecithin were sliced and visually observed, they were noted to have similar cell structure and texture to the DATEM samples.

TABLE 22