Patent Description:
Due to health issues, sugar consumption has become very high on the health agenda. Thus, within EU, there is a target to reduce added sugars by a minimum of <NUM>% by <NUM> in different food applications. A lot of sugar replacement ingredients have been developed in the last years to avoid addition of sugar to food, including artificial sweeteners (polyols), natural sweeteners (inulin, oligofructose), which are mainly hydrolyzed by acids or by enzymes.

Besides the health issues, there is a general interest in keeping the content of added sugar in baked product as low as possible. The reasons include reducing the cost price of the baked product, avoiding impact of fluctuations in the price of sugar, obtaining the possibility of reducing the amount of salt added, etc. Also, there is a growing general skepticism against synthetic or perceived un-natural additives in the consumer group.

Baked products with a reduced or without a content of added sugar are already known:.

Still, none of the processes demonstrated in the prior art i) shows that it is possible to significantly reduce the proofing time and ii) results in a baked product that is as paltable as a traditional baked product having been baked with added sugar and which has a low content of individual mono- or disaccharides and a low total content of mono- and di-saccharides.

In the following, the combination of the three enzymes in step i) is referred to as a cocktail of three enzymes.

The present invention also relates to this cocktail of three enzymes, namely to a combination consisting of an alpha-amylase (EC <NUM>. <NUM>), which is thermo-labile and has activity in a range of from <NUM> to <NUM>, an amyloglucosidase (EC <NUM>. <NUM>), which is thermo-stable and has optimum activity at about <NUM>-<NUM>, and a maltogenic amylase (EC <NUM>. <NUM>), which has optimal activity in a range of from <NUM> to <NUM>, and wherein the thermo-labile alpha-amylase, the thermo-stable amyloglucosidase and the maltogenic amylase are present in ratios corresponding to.

In embodiments, the present invention relates to a process for producing a baked product with no-added sugar, the process comprising.

In embodiments, the present invention also relates to a process for producing a baked product with no-added sugar, the process comprising.

In embodiments, the present invention also relates to a process for producing an un-baked product with no-added sugar. In such cases, the consumer will buy the un-baked product in frozen form and arrange for the final baking. Thus, the process comprises.

In aspects of the invention, the pre-baking step is included in the process. In other aspects both the baking step is included in the process.

Thus, in its broadest aspect, the present invention relates to a process for producing a product with no-added sugar, the process comprising.

In aspects of the invention, steps i), ii), and v) are included, where step v) is a baking step the final product is a baked product).

In aspects of the invention, steps i), ii), and v) are included, where step v) is a pre-baking step the final product is a pre-baked product).

In aspects of the invention, steps i), ii), and v) are included, where step v) is a pre-baking step followed by step iv) (the final product is a frozen, pre-baked product).

In other aspects of the invention, steps i), ii), iii) and v) are included, where step v) is a baking step (the final product is a baked, shaped product).

In other aspects of the invention, steps i), ii), and iv) are included (the final product is a proofed, frozen, un-baked product).

In other aspects of the invention, steps i), ii), iii) and iv) are included (the final product is a proofed, shaped, frozen, un-baked product).

Described herein are also the products obtained by the processes described above (not part of the claimed invention).

Important features of a process according to the invention are a) no addition of sugar and b) the use of a two-step enzymatic process.

In the present context the terms "no addition of sugar" and "with no-added sugar" mean that none of the ingredients used in the production of a baked product according to the present invention is a sugar in the form of a monosaccharide such as e.g. glucose, fructose, or a disaccharide such as e.g. maltose or saccharose, i.e. there is no external addition of sugar in the baking process. Other ingredients, such as flour may contain oligosaccharides or polysaccharides that enzymatically can be degraded to mono- or disaccharides and they may contain a minor amount of a mono- or disaccharide, such as about <NUM>-<NUM>% by weight of eg glucose, fructose, sucrose and raffinose. However, a person skilled in the art will know that the amounts of mono-, di-, oligo- and polysaccharides in a flour may vary dependent on the particular flour used.

One of the challenges in restraining from adding sugar to the dough is how to obtain sufficient and optimized proofing of the dough. Proofing of the dough is normally obtained by a fermentation process; whereby yeast organisms consume sugar in the dough and produce ethanol and carbon dioxide as waste products. The carbon dioxide forms bubbles in the dough and expands it (proofing).

When no sugar is added to the dough as in a process of the invention, another mechanism must apply. To this end, it is important that the flour contains a certain amount of damaged starch. Damaged starch refers to the portion of kernel starch that has been physically broken or fragmented during milling. Damaged starch is believed to have a strong influence on the dough and baking process. In the present process, damaged starch is a suitable substrate for the alpha-amylase to provide the necessary sugar molecules that are required for the fermentation process by the yeast. It is envisaged that the alpha-amylase (most likely with some contribution from the combination of amyloglucosidase and maltogenic amylase) relatively fast produces the necessary sugars and in a sufficient amount in to order to obtain fast proofing. It is assumed that the sugars formed are consumed approximately at the same time as they are formed. This is supported by the observation done by the inventors that the proofing time is reduced for dough with no-added sugar compared to dough with sugar added.

<FIG> show that the proofing step is controlled either with yeast or with proofing time, or both. The upper graph in both figures relates to a product without any sugar added. The other graphs relate to products having <NUM>% sugar added. In order to obtain e.g. a proofing height of <NUM>, the proofing time can be reduced from the proofing time from <NUM> to <NUM>, thus, in general a <NUM>% reduction in proofing time. Moreover, the choice and content of yeast can control the proofing stage and volume. In the examples Danish standard yeast (Malteser yeast) has been used. In general, a reduction in proof time of from <NUM> to <NUM>% is obtained. Experimental details are given in Example <NUM>.

Another observation made by the inventors is that it seems as if the presence of the combination of amyloglucosidase and maltogenic amylase also have impact on this first enzymatic step, as the release of sugars seems to be faster when all three enzymes are present compared with the presence of alpha-amylase alone. Thus, in this first step, both amyloglucosidase and maltogenic amylase may contribute to the release of sugar, even if they have a lower relative activity at the proofing temperature compared with the relative activity at higher temperatures (eg at about <NUM>). As seen from <FIG> all three enzymes (thermolabile amylase, thermostable amyloglucosidase and maltogenic amylase) contribute to the proofing of the dough. This observation supports the hypothesis by the inventor that even if the amyloglucosidase used is thermostable, it has some activity at proofing temperature, which normally is in a range of from about <NUM> to about <NUM>. The same applies to the maltogenic amylase.

An envisioned scenario could be that the maltogenic amylase contributes to the release of maltose from starch and that some of the maltose released from the starch by action of alpha-amylase and (maybe) by action of maltogenic amylase is further degraded to glucose by action of the amyloglucosidase present in the combination.

Another challenge in restraining from adding sugar to the dough is how to obtain a baked product that is palatable. To this end, taste, smell, aroma, consistency etc. of the baked product influence whether a consumer finds the baked product palatable. As seen from the examples herein, it has been possible to obtain palatable baked products without adding of sugar to the dough. As demonstrated in the examples herein, the process of the invention leads to baked products with a low content of mono- and di-saccharides. Thus, even if the sugar content is low, the baked product is palatable. The total concentration of mono- and di-saccharides (measured as fructose, glucose, lactose, maltose and saccharose) is at the most about <NUM>% w/w of the baked product, and the concentration of the individual sugar is:.

The concentration of individual sugar is based on the total weight of the baked product.

The present invention also relates to baked product with the above-mentioned contents of sugars. Baked products with a low content of sugars are highly relevant for many consumers. According to a recent survey <NUM>% of consumers both in Europe and in the USA reported that they are trying to reduce their sugar consumption.

As mentioned above, a cocktail of three enzymes is used in a process of the present invention. In the following is given details regarding the enzymes including the activity of the enzymes. However, it is to be borne in mind that the activities stated are measured in in vitro systems under standardized conditions (temperature, pH, humidity etc.). The activity of an enzyme is dependent on the conditions under which the enzyme is present. Thus, the activity in a dough and during baking may differ from the in vitro activities given herein.

The alpha-amylase used (EC <NUM>. <NUM>) is an enzyme, which hydrolyzes the degradation of alpha-<NUM>,<NUM>-glucosidic bonds in oligo- or polysaccharides such as in starch to yield maltose, but it does not act on maltose itself. Intermediate oligosaccharides such as dextrins are formed in the process. Alpha-amylase is an endoglucosidase which cleaves an internal glucosidic bond within an oligo- or polysaccharide.

The alpha-amylase may be an alpha-amylase of fungal or bacterial origin. Preferred is an alpha-amylase of fungal origin. The fungal origin may be from Aspergillus, such as Aspergillus oryzae, Aspergillus niger or Aspergillus kawachii. Examples of commercially available compositions comprising alpha-amylases are FUNGAMYL™, including Fungamyl <NUM> SG and Fungamyl Prime BAN® (all from Novozymes, Denmark), MYCOLASE®, Bakezyme P180, Bakezyme P500 (DSM, Gist Brocades), Grindamyl A <NUM>, Grindamyl A <NUM>, Grindamyl A <NUM>, Grindamyl A <NUM> (from IFF/Dupont) and Veron M4 /from AB Enzymes). Alpha-amylases of bacterial origin suitable for use in the present invention includes Biobake <NUM> (Kerry Ingredients), BAN <NUM> (Novozymes), Bakezymes AN <NUM> (DSM) and Grindamyl Max life (IFF/Dupont.

Especially suitable for use in the present invention is an alpha-amylase, which is a fungal alpha-amylase that is an endo-amylase that hydrolyzes (<NUM>,<NUM>)-alpha-D-glucosidic linkages in starch polysaccharides, and the fungal alpha-amylase is obtained from Aspergillus oryzae.

Alpha-amylases are normally used in bread inter alia to improve brown curst colour, to ensure fine and uniform crumb structure and/or to increase the volume of the bread.

However, in the present invention, an important feature of the alpha-amylase is its ability to degrade starch to provide mono- and disaccharides for use in the yeast fermentation process.

As it appears from the examples herein, a suitable alpha-amylase for use in the present invention is contained in the commercial product Fungamyl® <NUM> SG from Novozymes, Copenhagen. Other alpha-amylases have also been tested and found suitable for use. These include the enzymes Fungamyl from Novozymes, Grindamyl from IFF/Dupont, Bakezyme from DSM and Veron M4 from AB Enzymes.

Fungamyl® <NUM> SG contains alpha-amylase from Aspergillus oryzae. It has an activity of <NUM> FAU-F/g. According to a datasheet from Novozymes, the enzyme is yellow to light brown and appears as a granulate having a particle size of approx. <NUM>-<NUM> microns; it has an approx. density of <NUM>/ml. ; it is readily soluble in water at all concentrations that occur in normal usage. Other alpha-amylases having the same characteristics, or characteristics that deviate at the most <NUM>% from the characteristics mentioned above are contemplated to be suitable for use in the present invention; fx: one characteristic is the activity - this may be within a range of from <NUM> - <NUM> FAU-F/g; the particle size may be from <NUM> - <NUM> micron and the density may be from <NUM>-<NUM>/ml. FAU-F is a measure for enzyme activity. FAU refers to Fungal Alpha-amylase Unit, ie the amount of enzyme which breaks down <NUM> starch per hour at Novozymes' standard method for determination of alpha-amylase. Tests for alpha-amylase activity are well known in the art. See for example test described on Sigma Aldrich, or in <NPL>. Further methods are published in Enzymology, and the selection and application of methods are within the skill of a person skilled in the art.

The alpha-amylase has an activity in a range of from <NUM> to <NUM> (relative activity at least <NUM>% of maximal activity) and about <NUM>% relative activity at a temperature of <NUM> to <NUM>. Likewise, it has an optimum activity at a pH in a range of from <NUM> to <NUM> (more than <NUM>% relative activity) and about <NUM>% relative activity at about pH <NUM> to about <NUM>. At temperatures from approx. <NUM>-<NUM>, the activity decreases and at approx. <NUM>, the enzymes is <NUM>% inactivated. The alpha-amylase begins rapidly to be inactivated at temperatures greater than about <NUM>, i.e. before the starch gelatinizes (at about <NUM>) and is believed to have no or only little initial activity during the baking at about <NUM> to about <NUM>. Therefore, the alpha-amylase is believed to have no (or only little) contribution to the final sugar content in the baked product.

The commercial product, Fungamyl® <NUM> SG, contains approximately <NUM>% w/w alpha-amylase <NPL> (defined as enzyme concentration on dry matter basis), approximately <NUM>% w/w wheat flour <NPL>, approximately <NUM>% w/w wheat starch <NPL>, approx. <NUM>% w/w of water <NPL> and approximately <NUM>% w/w of dextrin <NPL>.

When Fungamyl® <NUM> SG is employed, it is normally used in an amount in a range for from about <NUM> to about <NUM> ppm/kg flour such as in a range of from about <NUM> to about <NUM> ppm/kg flour or in a range of from about <NUM> to about <NUM> ppm/kg flour such as about <NUM> ppm/kg flour. If another alpha-amylase is used a person skilled in the art will know how to calculate a suitable amount based on the activities given for Fungamyl® <NUM> SG and the other alpha-amylase used.

As used herein for alpha-amylase such as Fungamyl® the following applies:.

A thermolabile alpha-amylase is typically used in an amount corresponding to a range of from about <NUM> to about <NUM> Fau/kg flour such as from about <NUM> to about <NUM> Fau/kg flour.

Other commercial products may contain the same alpha-amylase or another alpha-amylase suitable for use in the present invention. Such products are also contemplated to be suitable for use in the present invention.

Compared with the Fungamyl® 4000SG product, a suitable alpha-amylase composition may be a composition, wherein there may be a variation in the content of flour (another flour than wheat may be used), there may be a variation in the concentration of flour (another concentration than <NUM>% by weight may be used) etc. Thus, a composition comprising an alpha-amylase suitable for use in the present invention may comprise:.

The above-given compositions are only examples of suitable compositions. Other compositions may also be suitable provided that they contain an alpha-amylase suitable for the present use. In general, such compositions contain one or more ingredients that make the enzyme stable for storages or that enable easy handling of the enzymes. A suitable composition may be in solid form or it may be in the form of a liquid.

Minor amounts of trace elements from the production process may be present in a composition. Normally, not more than a few percent at maximum is present.

Glucoamylase (<NUM>,<NUM>-alpha-D-glucan glucohydrolase, EC <NUM>. <NUM>), also denoted amyloglucosidase, is an enzyme, which catalyses the release of beta-D-glucose from the non-reducing ends of starch or related oligo- and polysaccharides.

The glucose sweetness intensity obtained in the final baked product is believed to come mainly from the action of the amyloglucosidase and its ability to release glucose. Moreover, it is believed to participate in the Maillard reaction yielding a richer, golden crust of the baked product.

An amyloglucosidase for use in the enzymatic combination for use in the present invention is an enzyme, which has optimum activity at about <NUM>-<NUM> and which has almost no activity at temperatures exceeding <NUM>. It has about <NUM>% relative activity at a temperature of from about <NUM> to about <NUM>, the activity being measured at pH <NUM> after <NUM> incubation time at the relevant temperature.

Amyloglucosidases are normally used in the baking industry to obtain more colour.

As it appears from the examples herein, a suitable amyloglucosidase is GoldCrust 3300BG from Novozymes, Denmark. Other suitable amyloglucosidases are Grindamyl AG 1500C, FD48, plussweet G (all from IFF/Dupont), Bakezyme AG <NUM> and Bakezyme AG <NUM> (both from DSM) and AMG <NUM> BG (from Novozymes).

Gold Crust 3300BG is derived from Aspergillus niger. The activity of the enzyme is expressed in Amyloglucosidase Units/g (AGU/g measured under reaction conditions pH = <NUM>, temperature <NUM> and an incubation time of <NUM>. The enzyme activity is determined based on the release of glucose and calculated relative to an enzyme standard (<NPL>.

When Gold Crustl® 3300BG is employed, it is normally used in an amount in a range for from about <NUM> to about <NUM> ppm/kg flour such as in a range of from about <NUM> to about <NUM> ppm/kg flour or in a range of from about <NUM> to about <NUM> ppm/kg flour such as about <NUM> or <NUM> ppm/kg flour. If another amyloglucosidase is used a person skilled in the art will know how to calculate a suitable amount based on the activities given for Gold Crust® 3300BG and the other amyloglucosidase used.

As used herein for a thermo-stable amyloglucosidase such as Gold Crustl® the following applies:.

A thermostable amyloglucosidade is typically used in an amount corresponding to a range of from about <NUM> to about1650 AGU/kg flour such as from about <NUM> to about <NUM> AGU/kg flour.

The above-given composition is only an example of a suitable composition. Other compositions may also be suitable provided that they contain an alpha-amylase suitable for the present use. In general, such compositions contain one or more ingredients that make the enzyme stable for storages or that enable easy handling of the enzymes. A suitable composition may be in solid or liquid form.

A maltogenic amylase (EC <NUM>. <NUM>) is able to hydrolyze starch, amylose and amylopectin to maltose. A maltogenic amylase may be produced from bacteria such as Bacillus subtilis (Novamyl® <NUM> BG) or Bacillus stearothermophilus. The enzyme used in the Examples herein is from Bacillus subtilis.

Maltogenic amylases are normally used in the baking industry for improving softness.

The crust of bread forms through the Maillard reaction, which is a chemical reaction between sugars and amino acids that occur at high heat. Amino acids are abundant in the flour and are not a limiting factor, whereas the content of sugar is believed to be a limiting factor.

As demonstrated in the examples herein, the maltogenic amylase contained in the commercial product Novamyl® <NUM> BG has proved to be suitable in the present context. Other suitable maltogenic amylases are Grindamyl Max life P100, U4, E50, Powerfresh <NUM>, Powerfresh <NUM>, Powerfresh <NUM>, Powerfresh <NUM>, Powerfresh <NUM>, Powerfresh <NUM>, Powerfresh <NUM> (all from IFF/Dupont), Novamyl 3D, Sensea BG, Novamyl Rye, Novamyl Pro <NUM> BG and Novamyl Ro <NUM> BG (all from Novozymes), Bakemaster Master, Bakemaster Fresh XL, Bakemaster Man <NUM>, Bakemaster Alpha (all from DSM), Veron <NUM>, Veron AC, Veron BA, Veron Sort+, Veron ELS and Amylofresh (all from AB Enzymes).

Novamyl® <NUM> BG contains a maltogenic amylase obtained from Bacillus subtilis. It has an activity of <NUM> MANU/g. It appears as a light brown powder in the form of a free-flowing, low-dusting granulate having a mean particle size of approximately <NUM>-<NUM> microns. It has an approximate density of <NUM>% g/ml. It is readily soluble in water at all concentrations that occur in normal usage. Other maltogenic amylases having the same characteristics, or characteristics that deviate at the most <NUM>% from the characteristics mentioned above, are contemplated to be suitable for use in the present invention; e.g.: one characteristic is the activity - this may be within a range of from <NUM> - <NUM> MANU/g; the particle size may be from <NUM> -<NUM> micron and the density may be from <NUM>-<NUM>/ml.

MANU is Maltogenic Amylase Novo Units. One MANU is defined as the amount of enzymes that produces <NUM>µmol glucose per minute using maltotriose as substrate under reaction conditions: pH = <NUM>, temperature = <NUM>, incubation time = <NUM>. The enzymatic hydrolysis of maltotriose results in the release of glucose, which can be determined quantitatively using a hexokinase assay (<NPL>).

The maltogenic amylase has an optimal activity in a range of from <NUM> to <NUM> (relative activity at least <NUM>%) and about <NUM>% relative activity at a temperature of <NUM> to <NUM> and a pH of <NUM>. Likewise, it has an optimum activity at a pH in a range of from <NUM> to <NUM> (more than <NUM>% relative activity) and about <NUM>% relative activity at about pH <NUM> to about <NUM>. The effect of temperature on maltogenic amylase activity. Maltogenic amylase was incubated at different temperatures for <NUM> minutes at pH <NUM> using maltotriose as substrate.

The commercial product, Novamyl® <NUM> BG, contains approximately <NUM>% by weight of wheat flour, <NPL> <NUM>% by weight of sodium chloride <NPL>, <NUM>% by weight of maltogenic amylase (defined as enzyme concentration on dry matter basis) <NPL> and <NUM>% by weight of water <NPL>. For more details, reference is given to the data sheet issued by Novozymes and valid from <NUM>-<NUM>-<NUM>. Other commercial products may contain the same maltogenic amylase or another maltogenic amylase suitable for use in the present invention. Such products are also contemplated to be suitable for use in the present invention. Compared with Novamyl® <NUM> BG, a suitable maltogenic amylase composition may be a composition, wherein there is a variation in the content of flour (another flour than wheat may be used), there may also be a variation in the concentration of flour (another concentration than <NUM>% by weight may be used) etc. Thus, a composition comprising a maltogenic amylase suitable for use in the present invention may comprise:.

Minor amounts of trace elements from the production process may be present in a composition.

When Novamyl® <NUM> BG is employed, it is normally used in an amount in a range for from about <NUM> to about <NUM> ppm/kg flour such as in a range of from about <NUM> to about <NUM> ppm/kg flour or in a range of from about <NUM> to about <NUM> ppm/kg flour such as about <NUM> ppm/kg flour. If another maltogenic amylase is used a person skilled in the art will know how to calculate a suitable amount based on the activities given for Novamyl® <NUM> BG and the other maltogenic amylase used.

As used herein for a maltogenic amylase such as Novamyl -stable amyloglucosidase such as Gold Crustl® the following applies:.

A maltogenic amylase is typically used in an amount corresponding to a range of from about <NUM> to about <NUM> Manu/kg flour such as from about <NUM> to about <NUM> Manu/kg flour.

The enzymes also impart other beneficial properties to the final product. Thus, as seen from the examples herein, the final product has excellent properties with respect to crust colour, shape of the product, uniformity, cell size, cell wall, cell form and crumb colour.

The dough may also contain other enzymes such as thermo-stable alpha-amylases, lipases, xylanases etc. However, these enzymes do not contribute to the content of sugar in the final product nor to the proofing time observed.

The flour used in the present process can be of any origin, provided it contains the necessary amount of damaged starch. The flour may be wheat flour, whole wheat flour, heat-treated flour, cake flour, rye flour, sifted rye, oat flour, barley flour, triticale (bread) flour, rice flour, corn flour, potato flour, heat-treated flour, bleached flour, or mixtures thereof, and/or it may include tapioca starch, corn starch, potato starch etc..

As demonstrated in the examples herein, a flour suitable for use in the present invention is wheat flour. The wheat flour may be any suitable wheat flour for example, one or more selected from the group consisting of all-purpose flour, bread flour, German type <NUM> flour, Reform flour, Manitoba flour, durum wheat flour, wheat flour based on soft or hard wheat types, Emmer, spelt & cake flours, other commercially available wheat flour, and combinations thereof.

There are generally speaking four types for wheat flour. White flour is made from endosperm only of the wheat grain. Brown flour includes some of the grains's germ and bran, while whole grain or wholemade flour is made from the entire grain, including the bran, endosperm and germ. Germ flour is made from the endosperm and germ, excluding the bran. All four types are suitable for use in the present invention.

As mentioned above, the flour used must have a certain content of damaged starch. In the present invention a content of at least <NUM>% w/w (based on the total weight of the flour). Generally, the content of damaged starch is at the most <NUM>% w/w. Thus, a suitable content of damaged starch is in a range of from <NUM> to <NUM>% w/w or <NUM>% w/w or more, <NUM>% w/w or more, <NUM>% w/w or more, <NUM>% w/w or more, <NUM>% w/w or more, <NUM>% w/w or more or <NUM>% w/w or more. In the examples herein, flour is used, wherein the content of damaged starch is from <NUM> to <NUM>%, notably from <NUM> to <NUM>% w/w (based on the total weight of flour) is suitable. If flour is used in combination with a starch, the total content of damaged starch is as described above.

Various starches may also be used as supplement to the flour.

A yeast suitable for use in the present invention is any yeast conventionally used in yeast-raised bakery. Suitable yeasts include dry yeast, sugar stable yeast and normal yeast. An especially suitable yeast is Saccharomyces cervisae. In the examples herein Danish standard yeast (Malteser yeast) has been used.

If necessary, pH of the dough is adjusted to a pH in a range of from about <NUM> to about <NUM> such as from about <NUM> to <NUM> by addition of one or more pH-adjusting agents. The adjustment of pH may be made to ensure the desired activity of the enzymes.

The dough may also comprise other commonly used ingredients in a dough. Such ingredients are typically mixed together with the other ingredients during preparation of the dough. Suitable additives include one of more of:
emulsifiers, fibers (such as maltodextrins, polydextrose, inulins, etc.), triglyceride, fat, pH adjusting additives, proofing adjusting additives, shortening agents, dough strengtheners, flour improvers, other enzymes including such enzymes that strengthen the dough; oxidizing enzymes, hemicellulose, lipase, protease, combinations thereof, ascorbic acid, sodium chloride, preservation agent, chemical leaving agent and other commonly used ingredients in baked products.

Mixing can be done with any suitable methods including a continuous mixer system, a spiral mixer of a fork mixer.

After mixing all the ingredients, the dough obtained is left to proofing at suitable conditions. As mentioned above, the proofing time is markedly reduced compared with doughs having added sugar. The proofing time may be reduced by <NUM>% such as <NUM>% or even <NUM>%. The proofing is normally carried out at slightly elevated temperatures compared to room temperature. The temperature is normally in a range of from about <NUM> to about <NUM>, notably from about <NUM> to about <NUM> or from about <NUM> to about <NUM> and at a relative humidity in a range of from <NUM>-<NUM>% RH.

The dough may also be obtained using other methods such as sponge dough, straight dough, poolished dough, liquid sponge, CBP (Chorleywood bread process), long fermentation or freezing technology. A sponge dough is a two-step bread making process. In the first step a sponge is made and allowed to ferment for a period of time, and in a second step the sponge is added to the final dough's ingredients. A straight dough is a single-mix process of making bread. The dough is made from all ingredients, and they are placed together and combined in one kneading or mixing session followed by fermentation. The CBP (Chorleywood bread process) process allows the use of lower-protein wheats and reduces processing time. The dough may also be prepared in a stepwise manner. Such a stepwise manner could include a pre-step to soften the dough followed by addition of the enzymes and the proofing the dough. In general enzymes are not added in the pre-step, but there could be situations, where inclusion of one or more of the enzymes is beneficial for the end result of the final product.

The dough obtained after mixing and the dough obtained after proofing are also described herein.

Thus, the dough for use in the process of the invention can be.

The following notations are also used: i) thaw and serve (ready to eat after thawing), ii) thaw and bake (par-baked), iii) pre-proofed dough (freezer to oven), and iv) raw dough (should be proofed and baked).

Thus, bake-off products are also objects of the present invention, ie proofed dough that has been baked to a certain extent, but the product needs further baking before intake thereof (pre-baked products).

The dough of the present invention contains flour with a content of damaged starch in a range of from <NUM>% w/w or more (based on the total content of flour) such as from <NUM> to <NUM>% w/w, from <NUM> to <NUM>% w/w or from <NUM> to <NUM>% w/w, a thermo-labile alpha-amylase and a composition containing a thermo-stable amyloglucosidase and a maltogenic amylase; a yeast; water, and optionally other ingredients common for preparing a dough. The dough contains no added sugar. The content of damaged flour is described above. In some cases, the dough may also contain one or more fibers such as maltodextrin or inulin. The content of the enzymes has been described herein before.

Before or after proofing, the dough may be shaped into the desired form. It may be subject to freezing, which normally involves a pre-freezing step at a temperature of from about -<NUM> to - <NUM> such as about -<NUM> for a time period of from about <NUM> to about <NUM> such as from about <NUM> to about <NUM> followed by stable freezing at about -<NUM>. The frozen product may be thawed before baking or directly placed in an oven for baking.

The freezing step is normally carried out when the product is sold as a pre-proofed or non-proofed product, i.e. the consumer smust baked the product themselves. For the non-proofed product, the consumers must both proof and bake the product themselves.

The dough may be frozen so that the consumer only needs to bake the dough in order to obtain the baked product. Thus, the present invention also relates to the dough obtained after proofing and wherein the dough has been frozen. When the frozen dough is baked it results in a baked product as described in the following. Freezing of the dough is typically used for laminated doughs.

The proofed product is baked or otherwise handled to obtain the final product. The baking is typically carried out at a temperature in a range of from about <NUM> to <NUM> or from about <NUM> to <NUM>, and the core temperature of the product is from <NUM> to <NUM>, from <NUM> to <NUM>, from <NUM> to <NUM> or from <NUM> to <NUM>. Instead of baking, the dough may be subjected to steam and hence, the product is obtained as a steambread. The steaming is normally carried out at a temperature of about <NUM>, but the core temperature of the product is the same as if baking has been employed. The dough may also be subject to proofing followed by cooking and baking. Thus, the step of baking may be replaced by a step of steaming.

At the beginning of the baking process, the temperature of the dough is close to room temperature and as the temperature of the dough rises when placed in the oven, the alpha-amylase will become inactive (or much less active) and the yeast will be inactivated. During this process, the combination of the amyloglucosidase and the maltogenic amylase is active. Whereas the first enzymatic step provided sugar as feed for the yeast, this second step provides sugar to the baked product so that the consumer finds it palatable, tasty, having a distinct pleasant flavour and texture. Therefore, all enzymes are not active (or equally active) at the temperatures, where mixing and proofing take place. Not all sugars that can be produced under optimal conditions for all enzymes present are produced during mixing and proofing as - if this was the case - all sugar content or most of it could be used by the yeast, leaving no particular sugar content in the final baked product. Accordingly, the alpha-amylase and the two enzymes in the combination of amyloglucosidase and maltogenic amylase have different activity patterns at different temperatures.

The baking results in a baked product. Such a product has a total content of mono- and di-saccharides that is higher than that in the ingredients making up the dough. Moreover, as seen from the Examples herein, a softer bun and more crispy croissants are obtained.

The invention also relates to a baked product obtained by the process set out in the appended claims comprising.

Notably, the total concentration of mono- and di-saccharides (measures as fructose, glucose, lactose, maltose and saccharose) is at the most about <NUM>% w/w such as in a range of from <NUM> to <NUM>% w/w based on the total weight of the baked product.

Typically, the concentrations of the individual sugars are:.

Any combination of the concentrations mentioned for the content of the individual sugars in the above paragraphs apply and is within the scope of the present application. Thus, the baked product may obtain:.

As seen from <FIG>, <FIG> and <FIG>, the concentration of the individual sugars can also be based on the total amount of sugar in the baked product. Based on the total content of sugars in the product (notably the content of fructose, glucose, lactose, maltose and saccharose), the content of the individual sugars may be:.

As mentioned herein before, any combination of the content for the individual sugars in the above paragraphs apply and is within the scope of the present application.

A baked product obtained by the processes of the invention may be in the form of burger buns, sandwich bread, whole bread, bread, muffins, pretzels, rolls, tortillas, pizza, bagels, pitas, ciabattas, gluten-free, foccacias, baguettes, loaves, sandwiches, waffles, pan cakes, laminated dough, croissants, pastry puff, cookies and biscuits etc..

When the baking step is steaming, a baked product may be a steambread.

A baked product obtained by a process according the present invention may be consumed by humans suffering from fructose intolerance. Fructose intolerance may be hereditary fructose intolerance (HFI), which is an inborn error of fructose metabolism caused by a deficiency of the enzyme aldolase B. If fructose is ingested, the enzymatic block at aldolase B causes an accumulation of fructose-<NUM>-phosphate, which, over time, results in the death of liver cells. Symptoms of HFI include vomiting, convulsions, irritability, hypoglycemia, hemorrhage, and potential kidney failure.

The invention also relates to a synergistic combination of a thermolabile alpha-amylase, a thermostable amyloglucosidase and a maltogenic amylase, which combination - when used in a process for producing a baked product without any addition of sugar - results in a baked product that contains <NUM>% w/w or less fructose, notably <NUM>% w/w or less, the weight being based on the weight of the baked product. As described herein, it is contemplated that a synergistic effect of the enzymes present in the dough is obtained in the first enzymatic step. Thus, it is envisaged that not only alpha-amylase is responsible for providing sugar to the yeast to consume during fermentation, but there may also be contribution from the combination of amyloglucosidase and maltogenic amylase in such a manner that the maltogenic amylase, although not very active at room and proofing temperature, contributes to the release of maltose, and that the amyloglucosidase contributes to the release of glucose, e.g. from maltose.

Moreover, in those case where it is desired to add an amount of sugar, but in a reduced amount compared to what is normally used, it is possible to replace some of the sugar with the cocktail of enzymes according to the invention. Moreover, replacing an amount of sugar added to a dough by a cocktail of enzymes of the invention results in a dough and a baked product just as good as the product without any reduction in the sugar content. Thus, the cocktail of enzymes of the present invention may also be used in situations where the "no-added sugar" is not the aim, but where a reduction in added sugar is the aim.

The synergistic combination is used in a process according to the invention to obtain a baked product as described above.

The synergistic combination typically comprises the thermo-labile alpha-amylase, the termo-stable amyloglucosidase and the maltogenic amylase in ratios corresponding to.

Thus, as an example, if the intention is to use <NUM> of the synergistic combination/kg flour, then <NUM> of the combination should contain from <NUM> to <NUM> Fau or from <NUM> to <NUM> Fau of the thermo-labile alpha-amylase, from <NUM> to <NUM> Agu or from <NUM> to <NUM> Agu of the thermo-stable amyloglucosidase, and from <NUM> to <NUM> Manu or from <NUM>-<NUM> Manu of the maltogenic amylase.

Likewise, if the intention is to use <NUM> of the synergistic combination/kg flour, then <NUM> of the combination should contain from <NUM> to <NUM> Fau or from <NUM> to <NUM> Fau of the thermo-labile alpha-amylase, from <NUM> to <NUM> Agu or from <NUM> to <NUM> Agu of the thermo-stable amyloglucosidase, and from <NUM> to <NUM> Manu or from <NUM>-<NUM> Manu of the maltogenic amylase.

The composition of the synergistic combination may be adapted to specific uses. Thus, e.g. for buns or toast, such a synergistic combination may contain the thermo-labile alpha-amylase, the termo-stable amyloglucosidase and the maltogenic amylase in ratios corresponding to.

A bread premix or a premix can be used to obtain a baked product. The premix typically contains flour, the enzyme combinationas described herein, and optionally other ingredients such as emulgators,sodium chloride, yeast, fibers, ascorbic acid, nuts, grains etc.). No added sugar is contained in the premix. The flour may be flour from grains such as wheat flour, corn flour rye flour, barley flour, oat flour, rice flour, sorghum, soy flour, and combinations thereof. The premix may be suitable for obtaining bread, buns etc. or for obtaining products based on laminated dough (such as e.g. croissants). When a product is made from the premix, water and yeast are added to the premix and the dough obtained is ready for proofing and baking.

All details and particulars described herein for one aspect of the invention apply mutatis mutandis to all other aspects of the invention and vice versa.

The following figures and examples are provided below to illustrate the present invention.

They are intended to be illustrative and are not to be construed as limiting in any way.

The analyses of the final baked products, relating to content of individual sugars and calories content were performed by Synlab, Malmo, Sweden. It is an accredited laboratory with No. <NUM> and ISO/IEC <NUM>.

The flour used in the examples all contain damaged starch in a concentration of <NUM>-<NUM>% w/w based on the total weight of the flour.

The enzymes: thermolabile alpha-amylase, thermo-stable amyloglucosidase and maltogenic amylase are used in the examples in amounts/kg flour as follows: <NUM> ppm/kg flour of thermo-labile alpha-amylase, <NUM> ppm/ka flour or <NUM> ppm/kg flour of thermo-stable amyloglucosidase and <NUM> ppm/kg flour of maltogenic amylase. The activity of the enzymes used can be calculated based on the text herein regarding the individual enzymes.

Two doughs (one without addition of sugar and the other with addition of sugar) were prepared from the following ingredients:.

All ingredients were mixed together in a mixer. Mixing a slow speed for <NUM> sec and at a high speed for <NUM> sec. A spiral mixer was used. The dough was then proofed for <NUM> to <NUM> at a temperature of <NUM> and a relative humidity of <NUM>%. After proofing the yeast was inactivated at <NUM> to <NUM> and baked at <NUM> to <NUM> for <NUM>.

The visual results are shown in <FIG>, where <NUM> denotes baked product from dough <NUM> and <NUM> denotes baked product from dough <NUM>.

Before baking, the two doughs were evaluated regarding stickiness, softness, extensibility, elasticity and dough temperature. No marked difference was found.

After baking, the bread was evaluated with respect to crust colour, shape of products, uniformity, cell size, cell wall, cell form and crumb colour. No difference was found.

Moreover, the content of individual sugars was evaluated with the following results:.

<FIG> shows the content of individual sugars given as a percentage of total amount of sugar.

As seen from the results above, there is a marked change in the content of the individual sugars, especially in the product with no-added sugar compared to the product with added sugar is the reduction low content of fructose, the increase in content of glucose and of maltose are noted. Moreover, in this example a minor reduction in total calories was seen.

Three doughs were prepared, one with a content of <NUM>% added sugar (B1), one with a content of <NUM>% added sugar(B2), and one without any added sugar (B3). The sugar content is based on the total amount of flour in the dough. The ingredients were as follows:.

All ingredients were mixed for <NUM>/<NUM> in a mixer, the dough is proofed at <NUM> and <NUM>% relative humidity for <NUM>, then the yeast was inactivated at <NUM>-<NUM> followed by baking of the dough at <NUM> to <NUM> temperature for <NUM>.

After mixing of the ingredients, the doughs were evaluated. All doughs were acceptable. The results were as follows:
<IMG>.

In general, adding of sugar gives a softer dough consistency. Without addition of sugar, the dough becomes less soft and stretchable. However, after baking all products had fine properties with respect to the following parameters.

The evaluation parameters are given below, and they also apply to the other Examples herein.

Regarding taste a lower sugar intensity was noted in the baked product without any added sugar compared to the product, where <NUM>% sugar had been added.

After baking the content of the individual sugars was determined. The following results were obtained - the results for the individual sugars are given as g/<NUM>:.

Two types of buns were prepared. One with the addition of <NUM>% sugar (A1) and the other without any addition of sugar. The doughs were prepared with the following ingredients:.

All ingredients were mixed for <NUM> sec/<NUM> sec in a mixer, the dough is proofed at <NUM> and <NUM>% relative humidity for <NUM>, then the yeast was inactivated at <NUM> C followed by baking of the dough at a temperature of <NUM>/<NUM> for <NUM>.

The appearance of the baked buns is illustrated in <FIG>.

It seems that the dough with added sugar is softer and more stretchable compared to the dough without added sugar. However, these properties do not adversely affect the processing of the dough.

After baking, the evaluation of the buns gave the following results:
<IMG>.

As is seen from the table above, only minor differences were observed. Without addition of sugar, there was a tendency to a more open crumb and a lighter colour. The taste without added sugar is less intense compared to the buns with <NUM>% added sugar.

The content of the individual sugars was measured with the following results:.

As seen from the results above, there is a marked change in the content of the individual sugars, especially in the product with no-added sugar compared to the product with added sugar a reduction low content of fructose, a decrease in the content of glucose and an increase in the content of maltose are noted. Moreover, in this example an approx. <NUM>% reduction in total calories was seen.

Products based on the following ingredients were prepared.

The dough and the baked product were made as described in Example <NUM>.

The dough was made from the following ingredient. The thermolabile alpha-amylase, amyloglucosidase and maltogenic amylase tested were added in amounts corresponding to those used in Example <NUM> or <NUM>.

The enzymes tested were: thermolabile alpha-amylase (Fau), thermostable amyloglucosidase (Gluco), maltogenic amylase (Manu).

The results are shown in <FIG>. The left-hand figure shows a synergistic effect when alpha-amylase and amyloglucosidase is combined and the volume after proofing is increased from <NUM> (no sugar added) to <NUM> (i.e. <NUM>%) or from <NUM> (when <NUM>% sugar was added to the recipe) to <NUM> (i.e. <NUM>%). The volume index, when dough with no enzymes and no sugar is <NUM>, is as follows:.

The right hand figure shows that addition of alpha-amylase or amyloglucosidase as single enzymes gives increased volume after proofing compared to dough with no enzymes added and either having no sugar added or <NUM>% sugar added. When all three enzymes are added, the best result regarding volume is achieved. The volume index, when dough with no enzymes and no sugar is <NUM>, is as follows:.

As seen from the table above, the doughs containing enzymes are better than the dough without enzymes and sugar and better or alike the dough containing <NUM>% sugar and no enzymes.

After baking the baked products obtained from doughs containing enzymes are better than the baked product obtained from dough without enzymes and sugar and better or alike the baked product obtained from dough containing <NUM>% sugar and no enzymes. The columns in the table above are the same as in the previous table.

Crossaints were made based on the following recipe:.

Croissants were made with no-added sugar, but with content of thermolabile alpha-amylase, thermostable amyloglucosidase and maltogenic amylase. The results of the baked croissants are shown in <FIG> (reference) and 8B (no-added sugar).

Four doughs with no added sugars were made and compared with standard. All baked products resulting from doughs with no sugar added had good as good as or better volume, structure, taste, sugar flavor, color compared with the croissants with <NUM>% sugar content.

In an internal triangle test with <NUM> participant only one respondent noticed the difference.

This example illustrate that using the combination of enzymes as claimed herein also can replace <NUM>-<NUM>% of added sugar without any lack of quality.

The results show that the combination of enzymes of the invention can replace <NUM>-<NUM>% of sugar without any lack of quality, use of the combination of enzymes gives better softness and freshness compared to benchmark and the addition of fiber gives shorter (dry) bide compares to benchmark. <FIG> shows the result - from left to right: Test <NUM>-Test <NUM>.

Toast and bun doughs with different content of sugar and with or without the enzyme cocktail of the invention were tested.

The results are shown in <FIG> and <FIG>. As seen from <FIG> an increase in height after <NUM> is about <NUM>% for the dough with the three enzymes compared with about <NUM>% for the doughs containing <NUM>% or <NUM>% sugar. After <NUM> the increase is about <NUM>% for the dough with the three enzymes compared with about <NUM>% for the doughs containing <NUM>% or <NUM>% sugar. After <NUM> the increase is about <NUM>% for the dough with the three enzymes compared with about <NUM>-<NUM>% for the doughs containing <NUM>% or <NUM>% sugar. In order to gain twice the height compared with the start value, a toast dough containing the combination of the three enzymes accoding to the invention will reach this at least <NUM> faster than that obtained for dough with <NUM>%, <NUM>%, <NUM>% sugar without the combination of enzymes.

The results are shown in <FIG> and <FIG>. As seen from <FIG> an increase in height after <NUM> is about <NUM>% for the dough with the three enzymes compared with about <NUM>%-<NUM>% for the doughs containing <NUM>% or <NUM>% sugar. After <NUM> the increase is about <NUM>% for the dough with the three enzymes compared with about <NUM>%-<NUM>% for the doughs containing <NUM>% or <NUM>% sugar. After <NUM> the increase is about <NUM>% for the dough with the three enzymes compared with about <NUM>%-<NUM>% for the doughs containing <NUM>% or <NUM>% sugar. In order to gain twice the height compared with the start value, a bun dough containing the combination of the three enzymes accoding to the invention will reach this at least <NUM> faster than that obtained for dough with <NUM>% or <NUM>% sugar without the combination of enzymes.

This example illustrates that it is possible to replace some of the sugar with the enzyme combination according to the invention and obtain products with lower content of fructose and higher content of glucose and maltose. Addition of <NUM>% maltodextrin does not markedly change the content of sugars compared to the buns with the enzyme combination.

Claim 1:
A combination consisting of an alpha-amylase (EC <NUM>.<NUM>), which is thermo-labile and has activity in a range of from <NUM> to <NUM>, an amyloglucosidase (EC <NUM>.<NUM>), which is thermo-stable and has optimum activity at about <NUM>-<NUM>, and a maltogenic amylase (EC <NUM>.<NUM>), which has optimal activity in a range of from <NUM> to <NUM>, and wherein the thermo-labile alpha-amylase, the thermo-stable amyloglucosidase and the maltogenic amylase are present in ratios corresponding to
from <NUM> to <NUM> Fau or from <NUM> to <NUM> Fau of the thermo-labile alpha-amylase,
from <NUM> to <NUM> Agu or from <NUM> to <NUM> Agu of the thermo-stable amyloglucosidase, and
from <NUM> to <NUM> Manu or from <NUM> to <NUM> Manu of the maltogenic amylase.