Use of hyaluronan in bakery products

Description

The present invention covers the use of hyaluronan as additive in foods, specifically in bakery goods and dough products.

The quality of bakery goods is influenced by a number of factors, such as, for example, the raw materials and formulations used, processing and the baking methods used.

Hyaluronan is a naturally occurring, unbranched, linear mucopolysaccharide (glucosaminoglycan) which is composed of alternating molecules of glucuronic acid (GIcA) and N-acetylglucosamine (GIcNAc). The basic building block of hyaluronan consists of the disaccharide glucuronic acid-beta-1 ,3-N-acetylglucosamine. This repeat unit is joined together in hyaluronan by beta-1 ,4 linkages.

In the field of pharmacy, the term hyaluronic acid is often used. Since hyaluronan is present mostly as polyanion and not as free acid, the term hyaluronan is preferably used below, where both molecular forms are deemed to be covered by the respective designation.

In cosmetic products, in particular in skin creams and lotions, hyaluronan is often used as a moisturizer on account of its high water-binding capacity.

Furthermore, preparations comprising hyaluronan are sold as so-called food supplements (nutraceuticals), which are also used for animals (e.g. dogs, horses) for preventing and alleviating arthrosis. Hyaluronan has extraordinary physicochemical properties, such as, for example, properties of polyelectrolytes, viscoelastic properties, a high water-binding capacity, properties of gel formation which, besides further properties of hyaluronan, are described in an overview article by Lapcik et al. (1998, Chemical Reviews 98(8): 2663-2684).

The extraordinary properties of hyaluronan open up a multitude of possibilities for use in highly diverse fields, such as, for example, pharmacy, the cosmetics industry, in the production of foods and animal feeds, for technical applications (e.g. as lubricants) etc.

The most important applications in which hyaluronan is currently used are in the medical and cosmetic sector (see e.g. Lapcik et al., 1998, Chemical Reviews 98(8): 2663-2684; Goa and Benfield, 1994, Drugs 47(3): 536).

A detailed depiction of the chemical structure of hyaluronan and its possible uses is given, for example, in WO 2006/032538.

A use of hyaluronan in foods has hitherto been described for the "dairy products" sector, i.e. for quark foods, yoghurts, cheese and other milk products (WO 2004/089098, WO 2005/058053) or as additive in beverages or for snacks (WO 2004/004686, WO 2004/002423). WO 2005/058068 describes the use of Hyaluronan as texturizer in connection with an other texturizer, e.g. starch, mainly for the production of foods or - products respectively, when the temperature of the food product is 50 to 100 ⁰ C.

An application which brings about both a higher water absorption in doughs and also improved digestibility as well as an improved "anti-staling" of the hence produced bakery products has hitherto not been described.

The present invention covers a composition comprising wheat flour in combination with hyaluronan in an amount which brings about a higher water absorption of the dough compared to compositions which do not contain hyaluronan.

Within the scope of the invention hyaluronan was utilized as an additive to the used flour.

During baking processes, baking losses usually arise, which are understood by the person skilled in the art as meaning the weight loss of the dough and/or of the dough items during baking. In the first instance, this is evaporating dough water and minimally other volatile constituents such as alcohol, organic acids and esters; the person skilled in the art is therefore just as likely to speak of "water loss".

Water losses during the baking process have a disadvantageous effect on the freshness of the bakery goods, which age earlier as a result, i.e. become "stale". This in term adversely affects the taste of the bakery goods and thus the so-called "mouth feel".

There is therefore a great need for methods in which the bakery goods and dough products have a relatively high water content after baking. As a result, it is possible to reduce the baking losses during the production of dough goods and particularly to achieve properties such as improved taste and an improved mouth feel, and also to increase the baking yield.

The present invention provides a composition which comprises wheat flour in combination with hyaluronan in an amount which brings about a higher water absorption of the dough compared to compositions which do not contain hyaluronan.

In a particular embodiment, the water absorption is increased when the composition according to the present invention consists of a combination of wheat flour with low molecular weight hyaluronan (LMW). In the case of this combination, the water absorption is increased by 0.5 to 25%, preferably by 1.0 to 22%, particularly preferably by 1 .5 to 20%, very particularly preferably by 2 to 15%, extraordinarily preferably by 3 to 12% and very extraordinarily preferably by 5 to 10%.

Low molecular weight hyaluronan (LMW-HA) is understood by a person skilled in the art as meaning a hyaluronan with a weight between 0,1 and 0.7 MDa. The present invention encompasses the use of low molecular weight hyaluronan, whose molecular weight may have any value between 0,1 and 0.7MDa, preferably a molecular weight of 0,2 to 0,65 MDa, particularly preferably of 0,3 to 0,6 MDa and very particularly preferably of 0,4 to 0,55 MDa.

In contrast to this, high molecular weight hyaluronan (HMW-HA) is understood by the person skilled in the art as meaning a hyaluronan with a molecular weight between 0,9 to 3 MDa. The present invention encompasses the use of high molecular weight hyaluronan, whose molecular weight may have any value between 0,9 to 3MDa, preferably a molecular weight of 1 ,0 to 2,5 MDa, particularly preferably of 1 ,5 to 2MDa and very particularly preferably of 1 ,7 MDa.

The hyaluronan used in the present invention can originate from various sources. It may either be a commercial hyaluronan which is of animal or microbial origin or Hyaluronan which of vegetable origin.

In connection with the present invention, the term "bakery goods" is then taken to mean a genenc term for dough items which may be in varying "states", namely unbaked, prebaked or ready-baked

The composition according to the invention is furthermore notable for the fact that it comprises 0 1 to 6% hyaluronan In a further embodiment, it comprises 0 1 to 5.5%, preferably 0.1 to 5%, particularly preferably 0 2 to 4 5%, very particularly preferably 0.3 to 4%, extraordinarily preferably 0.4 to 3.5%, very extraordinarily preferably 0 5 to 3% and very very extraordinarily preferably 1 to 2 % hyaluronan

In a further embodiment the composition according to the invention is notable for the fact that the hyaluronan utilized is added as powder to the flour

In a particular embodiment the composition according to the invention is notable for the fact that the hyaluronan utilized is added in dissolved condition to the flour. Therefore, the hyaluronan is resolved into water and than added to the flour for the production of the dough. The concentration of the hyaluronan in the dough is 0,5 and 1 ,0%, respectively.

The composition according to the invention is also notable for the fact that it brings about a higher dough yield Dough yield is understood by the person skilled in the art as meaning the dough weight based on 100 parts of flour after it is mixed with water, which is given in %. Surprisingly, it has been found that the dough yield is increased by 1 to 15% compared to the use of a composition without the addition of hyaluronan.

In a particular embodiment, the composition is notable for the fact that the hyaluronan present is low molecular weight hyaluronan (low molecular weight = LMW)

In a particular embodiment, the dough yield is increased especially for the combination of wheat flour with low molecular weight hyaluronan (LMW-HA), by 1 to 15%, preferably by 1 to 10%, particularly preferably by 2 to 8% and very particularly preferably by 3 to 5% compared to the use of a composition without hyaluronan

Surprisingly, it has been found that resolving of the hyaluronan in water prior to adding

to the flour effects a clear increase of the dough yield. In a particular embodiment with pre-resolved Hyaluronan the dough yield is increased by 1 to 20%, preferably by 1 to 16%, particularly preferably by 2 to 10% and very particularly preferably by 4 to 8%. The use of 0,5% pre-resolved high molecular weight hyaluronan causes an increase of the dough yield of 2%, the use of 0,5% pre-resolved low molecular weight hyaluronan an increase of even 4%.

Furthermore, it was surprising that when using the composition according to the invention, the bread yield is increased by 0,5 to 12%.

In connection with the present invention, bread yield is to be understood as meaning the weight of the attained bakery goods based on 100 parts of flour (declared in %).

In a further preferred embodiment, when using powder-low molecular weight hyaluronan as additive the bread yield is increased by 1 to 10%, preferably by 2 to 8%, particularly preferably by 3 to 5% and very particularly preferably by 4 to 5,5%. High molecular weight hyaluronan powder as additive does not lead to an increase of the bread yield.

In a further preferred embodiment when using pre-resolved low molecular hyaluronan as additive the bread yield is increased by 0,5 to 10%, preferably by 1 to 7% and particularly preferably by 2 to 5%.

In a further preferred embodiment when using pre-resolved high molecular hyaluronan as additive the bread yield is increased by 0,5 to 10%, preferably by 1 to 7% and particularly preferably by 3 to 5%.

The use of the composition according to the invention for achieving higher bread yields is likewise covered.

The features which are described above for the composition according to the invention are to be applied to the method according to the invention.

Methods to produce the bakery products according to the invention are a further matter of the invention.

The features which are described above for the composition according to the invention are to be applied to the bakery products according to the invention.

The composition of the invention covers groceries (foodstuffs and pleasing products), which contain the flour and other ingredients and substances the baker will use for the production of bakery products which will be used for the nutrition of the humans. Substances which are absorbed by the human for the use of feeding are e.g. dietary fibers, water, carbohydrates, proteins, fats, vitamins, secondary plant ingredients, mineral nutrients, flavours, flavours additives and/or food additives.

Furthermore, the present invention covers a method for achieving higher dough and bread yields wherein wheat flour in combination with hyaluronan as additive is used for the baking.

In a preferred embodiment, the method according to the invention is one in which the wheat flour used for the baking is used in combination with high molecular weight hyaluronan.

In a particularly preferred embodiment, the method according to the invention is one in which the wheat flour used for the baking is used with low molecular weight hyaluronan as additive.

Surprisingly, it has been found that when using low molecular weight hyaluronan (LMW-HA) in the method according to the invention, the bread yield was increased markedly. Here, the increase in the bread yield was 1 to 15%, preferably 1-10%, particularly preferably 2-8% and very particularly preferably 3-5%, compared to the use of a composition which does not comprise hyaluronan.

The method according to the invention is furthermore notable for the fact that a higher dough yield results therefrom. Here, the dough yield is increased by 0,5 to 15% compared to the use of a composition without the addition of hyaluronan. The dough yield is increased by 1 to 10%, preferably by 2 to 8% and particularly preferably by 3 to 5%, compared to the use of a composition without hyaluronan.

Surprisingly, it has been found that when using low molecular weight hyaluronan (LMW-HA) in the composition according to the invention, the bread volume was increased markedly.

"Volume yield" is another word for Bread volume and is measured in ml/10Og flour. The term "bread volume" is not limited to bread but encompassed the volume of all kind of bakery products.

By adding more than 0,5% low molecular weight hyaluronan as powder, the volume yield is increased by 10 to 20% compared to the use of a composition without the addition of hyaluronan. The dough yield is increased preferably by 10,5 to 18%, particularly preferably by 13 to 17% and very particularly preferably by 15 to 16,5%. Surprisingly, the use of only 0,5% pre-resolved low molecular weight hyaluronan resultes in an considerable increase of the bread volume. In this particular embodiment, the bread volume is increased by 1 to 15%, preferably by 2 to 10%, particularly preferably by 3 to 7,5% and very particularly preferably by 5 to 6,5%.

In a further embodiment, by adding 2% high molecular weight hyaluronan as powder the bread volume was increased by 1 ,2%.

In a further embodiment high molecular weight hyaluronan was pre-resolved and added to the flour. Surprisingly, already by the use of at least 0,5% pre-resolved high molecular hyaluronan the volume of the bakery products is increased by 0,5 to 10%, preferably by 0,6 to 8% and particularly preferable by 1 to 6% and very particularly preferable by 2 to 3,7%.

In a further embodiment, the present invention covers a bakery good which has been prepared from wheat flour, comprising hyaluronan as additive.

Surprisingly, it has also been found that the bakery goods which have been produced from wheat flour in combination with hyaluronan as additive have a reduced crumb strength.

When using low molecular weight hyaluronan, the crumb strength was reduced by 6 to

35%. This effect was evident particularly when more than 0.2% of LMW-HA were used. Preferably, the crumb strength was reduced by 8 to 30%, particularly preferably by 12 to

25% and very particularly preferably by 15 to 20%.

Surprisingly, by using pre-resolved low molecular weight hyaluronan the crumb strength was reduced by 5 to 40% compared to the control. In a preferred embodiment, one day after baking the crumb strength was reduced by 9 to 35%, particularly preferably by 9 to 30%. In a further embodiment 3 days after baking the crumb strength was reduced by 10 to 35%, preferably by 15 to 30%, particularly preferably by 15 to 35%.

The features which are described above for the bajery products according to the invention are to be applied to the composition and the method according to the invention.

Further surprisingly, the bakery products according to the invention show characteristics of "anti-Staling", which means that retrogradation is remarkably delayed.

For the expert, retrogradation is the process when starchgels and agglutinated starch become insoluble, which means that the starch passes into a soluble, (very) swollen status in to an insoluble. This process is irreversible and leads to a microcrystalline structure. The retrogradation is the reason for the so-called "frumpy" status (staling) of bread and bakery products. In this process the starch of the flour partly dispenses the physically bound liquid and passes into an crystalline status. The "frumpy" bakery product may have either a soft, often like sponge rubber or hard texture.

To slow-down the process of Staling, additives may be used, which e.g. as emulgators avoid the crystallization of the starch and therefore act as "anti-staling" components.

The characteristics of water in doughs or bakery products, respectively is not that well known, yet. Up to know it was guessed that hyaluronan binds the water of the dough in a way that the starch will be dehumidified and by that the "staling" is advanced.

Surprisingly by using hyaluronan as additive the water balance in the dough seems to be improved regarding the "anti-staling" characteristics of the bakery product.

In a further embodiment of the invention by using 0,5% pre-resolved low molecular weight hyaluronan the water uptake clearly increases. This is independent of the use of

low or high molecular weight hyaluronan. In this embodiment the water uptake is increased by 1 to 18%, preferably by 3,5 to 15,9 %, particularly preferably by 5,9 to 15,3% and very particularly preferably by 7,5 to 12%.

In a further embodiment by using pre-resolved high molecular weight hyaluronan the water uptake is increased by 1 to 12%, preferably by 2,1 to 11 % and particularly preferably by 2,1 to 9,4%.

By using this embodiment, the pre-resolved hyaluronan, only half of the used hyaluronan is needed compared to using hyaluronan as powder. Either low or high molecular weight hyaluronan may be used. The improvements of the bread volume and the "anti-staling" are maintained. When using hyaluronan as powder, only low molecular weight hayluronan displayes a comparable high water uptake.

In another aspect of the present invention, it was surprisingly found that consumption of these bakery goods brings about improved compatibility. This can also be referred to as improved digestibility and can be ascertained through the amount of released glucose. The release of glucose was 5-20% lower for bakery goods to which hyaluronan has been added than for bakery goods to which no hyaluronan has been added. In a preferred embodiment, the glucose release was 7-18% reduced, particularly preferably 9-17% reduced and very particularly preferably 10-15% reduced, compared to the use of a composition without hyaluronan.

A particular advantage of the composition and method according to the invention is the reduction in the fraction of rapidly digestible flour and/or starch since a rapid release of relatively large amounts of glucose and its absorption via the small intestine epithelium leads to an abrupt increase in the blood sugar level. This results in a release of insulin (insulin response). The continual consumption of foods with a high glycemic load and the insulin release associated therewith is suspected of being a risk factor in the development of diseases such as high blood pressure, obesity, heart diseases and type Il diabetes. An improved supply of bread and dough goods which, on account of their composition, bring about slower release of glucose would therefore be of great benefit in terms of health.

Material and methods

The following methods were used in the examples. These methods can be used for carrying out the processes according to the invention; they constitute specific embodiments of the present invention, but do not limit the present invention. The person skilled in the art is aware that he is able to carry out the invention in the same manner by modifying the described methods and/or by replacing individual parts of the method by alternatives.

1. Flour/flours used

A commercial baking wheat flour (wheat flour type 550 Meister / MM = "Meistermehl") from Kampffmeyer Mϋhlen (Werk Schϋttmϋhle Berlin, DE) was used. Depending on the charge the analytical and rheological data of the commercial flour may differ. Therefore the results of the experiments were compared with the flour as control accordingly.

Analytical data:

2. Hyaluronan

The commercial hyaluronic acids used were acquired via GfN from the manufacturer (CPN spol. s.r.o.., Dolπi Dobrouc, CZ).

Hyaluronic acid sodium salt 1.7: HMW (high molecular weight)

Powder, batch number: 3019510 Molecular weight: 1.7 MDa

Hyaluronic acid sodium salt 0.4-0.6 MDa: LMW (low molecular weight)

Powder, batch number: 30120602 Molecular weight: 0.55 MDa

Noncommercial hyaluronan: In addition, vegetable hyaluronan was used which, as described in WO 2006/032538, was isolated from tomatoes and characterized.

3. Water absorption and dough rheological investigations The analysis of the commercial flour with a low molecular weight and high molecular weight hyaluronan (LMW-HA and HMW-HA) for the water absorption and also the dough-rheological properties was carried out in accordance with the standard method of the International Association for Cereal Science and Technology (ICC/www.icc.or.at). This method can be found under Farinograph (ICC Standard 1 15/1 ).

The rheological characteristics of the flours were measured with an farinograph. By that the water uptake capacity (amount of water which was uptaken and needed to produce a dough of certain texture) and the texture of the dough (by the resistance of the dough subtended to the kneader) is measured. This resistance is transferd to a scale and will be noted in a farinogram (power-time-diagram). The farinogram shows:

- the time which is needed to get a certain texture of the dough

- the time in which the texture of the dough will stay stable

- the decrease of the texture in a certain range of time

Use of pre-resolved hyaluronan

In a further embodiment hyaluronan was dissolved in water and than added to the flour for making the dough. Either low molecular weight hyaluronan as well as high molecular weight hyaluronan were used. The hyaluronan powder was dissolved in the water ("Schuttwasser") which will be added to the flour to produce the dough. Finally the concentration of the hyaluronan in the dough is 0,5% or 1 ,0%.

4. Baking experiments:

The baking experiments were carried out both in the baking oven and also in a DSC (differential scanning calorimetry, PerkinElmer, USA) at Bayer BioScience GmbH (Potsdam, DE) in accordance with standard methods. For this, commercial baking wheat flour type 550 in each case with and without the addition of hyaluronan of differing molecular weights was used.

Figure 1 shows an overview of the baking processes carried out.

Compositions and methods of the baking experiments: Baking oven: baking-tin white bread (TWB):

Ingredients Contr.* +HA +HA +HA +HA + HA + HA

Flour 100 100 100 100 100 100 100

Hyaluronan HMW - - - 1 2

Hyaluronan LMW - 0,1 0,5 1 2

Salt 2.0 2.0 2.0 2.0 2.0 2.0 2.0

Yeast (fresh) 4.0 4.0 4.0 4.0 4.0 4.0 4.0

Cooking fat 5.0 5.0 5.0 5.0 5.0 5.0 5.0

Sugar 1.0 1.0 1.0 1.0 1.0 1.0 1.0

Water ³ var. var. var. var. var. var. var.

a Amount of added water is applied according to Farinogram data * Flour is set as 100 parts, the other components then added thereto.

Mixer: Farinograph kneading chamber (300 g of flour) from Brabender,

Duisburg/DE

Mix: Two minutes at speed (63 rpm) Two minutes (or more; according to Farinogram data) further at speed

63 rpm

Desired dough temperature is 26 ⁰ C. Resting: 10 minutes

Division: Divide dough into 250 g dough pieces, round using the Brabender rounding machine and shape lengthwise by hand.

Item refining: The shaped loaves are placed in baking tin molds and refined in the

fermenting cabinet for 60 minutes at 32 ⁰ C and 87% relative atmospheric humidity.

Baking: 30 seconds at 240 ⁰ C (with 80 ml vapor cloud)

2.00 min at 210 ⁰ C 27.30 min at 200 ⁰ C

DSC baking simulation (bread crumbs):

Ingredients Control ^* +HA

Flour 100 100

Hyaluronan LMW - 0.5

Salt 2.0 2.0

Yeast (fresh) 4.0 4.0

Cooking fat 5.0 4.0

Sugar 1.0 1.0

Water ³ variable variable a Amount of added water is applied according to Farinogram data * Flour is set as 100 parts, the other components then added thereto.

Mixer: 1 g of flour mini-spiral mixer (Pfeiffer)

Mix: 1 minute premixing at speed (? rpm)

5 minutes kneading at speed (? rpm) Resting: 5 minutes

DSC pan: 30-35 mg of dough are transferred to the DSC pan

DSC baking:

Baking profile: Heating from 20-110 ⁰ C (10°C/min) Cooling from 110-20 ⁰ C (10 ^β C/min)

The DSC pans consist of stainless steel with a diameter of 0 = 6 mm and height = 2 mm. The pans are closed, i.e. no crusts are formed.

5. Measuring the moisture The moisture (water content) of the bread sample is ascertained by means of the Karl- Fischer titration method (Karl-Fischer titrator AQUA 40 000, Analytik Jena Group, Jena, DE).

6. Crumb strength measurement

The crumb strength during storage (after 1 day and 3 days) was determined using a

Texture Analyzer (TA-XTplus, Stable Micro Systems, England). TA setting:

Test type: Pressure (compression)

Forwards rate: 2.00 mm/sec

Test rate: 0.50 mm/sec

Backwards rate: 0.50 mm/sec Target parameter: Path

Path: 7 mm

Trigger value: Auto (force)

Trigger force: 2.O g

Stop recording at: Start position

7. Determination of the fraction of resistant starch (digestibility)

The fraction of resistant starch is determined in accordance with the method described in Englyst et al. (Europ. J. of Clinical Nutrition 46 (Suppl. 2), (1992), pp. 33-50) (see in particular the following sections from Englyst et al., pages 35-36: "Reagents, Apparatus, Spectrophotometer"; pages 36-37: paragraph "Measurement of free glucose (FG)"; page 38: paragraph "Measurement of RDS and SDS").

The method from Englyst et al. can alternatively be carried out as described by Zhang et al. (Biomacromolecules 7, (2006): 3252-3258, in particular page 3253: Methods, Enzymatic Starch Hydrolysis).

On a laboratory scale, the method from Englyst et al. can be carried out in the following way with wheat flour.

To prepare the enzyme solution, 1.2 g of pancreatin (Merck) are extracted in 8 ml of water for 10 minutes at 37 ⁰ C. After centrifugation (10 min, 3000 rpm; RT), 5.4 ml of the supernatant are mixed with 84U of amyloglucosidase (Sigma-Aldrich, Taufkirchen) and topped up to a final volume of 7 ml with water. In parallel, 10 mg (dry weight) of wheat flour or wheat starch per sample are admixed in

a 2 ml reaction vessel with 0.75 ml of sodium acetate buffer (0.1 M sodium acetate pH 5.2; 4 mM CaCb) and incubated for 5 minutes at 37 ⁰ C to warm the mixture. Digestion of the starch is started by adding in each case 0.25 ml of enzyme solution per mixture. The control is a mixture to which water is added instead of enzyme solution. After 20, 60 and 120 minutes, aliquots of 100 μl are removed and added directly to four times the volume of ethanol, whereupon the enzymes are inactivated. This dilution is used for measuring the content of glucose.

For this, 2 μl of dilute sample are mixed with 200 μl of measurement buffer (100 mM imidazole/HCI pH 6.9, 5 mM MgCI ₂ , 1 mM ATP, 2 mM NADP) and the absorption of the sample is ascertained at 340 nm. The conversion of the glucose is started by adding 2 μl of enzyme mix (10 μl of hexokinase, 10 μl of glucose-6 phosphate dehydrogenase, 80 μl of measurement buffer) and the equimolar conversion of NADP to NADPH at 340 nm is monitored until a plateau is reached. The ascertained glucose amounts are compared with the initial weight and give the fraction of the sample which has been released as glucose after the corresponding period.

Description of the figures: Fig 1 : Overview of the Baking process Fig 2: Dough and bread characteristics using low molecular hyaluronan as powder and pre-resolved

Fig 3: Dough and bread characteristics using high molecular hyaluronan as powder and pre-resolved

Fig 4: Release of glucose upon digestion of the bread crumbs, comparison of the storage times with and without hyaluronan

Examples

1. Water absorption and dough-rheological properties

Following the addition of LMW-HA, a significant increase in the water absorption of the flour up to 10% is recorded.

Table 1: Farinogram data using different concentrations of low molecular weight (LMW) hyaluronan as powder

Upon using LMW-HA there is an increase in the water absorption of the dough.

Table 2: Farinogram data using different concentrations of high molecular weight hyaluronan (HMW-HA) as powder

When using HMW-HA, by contrast, a reduction in the water absorption of the dough is found.

Table 3: Farinogram data using different concentrations of pre-resolved low molecular weight hyaluronan (LMW-HA)

When using pre-resolved LMW-HA, an increase in the water uptake of the dough is found.

Table 4: Farinogram data using different concentrations of pre-resolved high molecular weight hyaluronan (HMW-HA)

When using pre-resolved HMW-HA, an increase in the water uptake of the dough is found.

2. Results of the baking experiments:

The results of the baking experiments are given below:

Table 5: Baking experiments using different concentrations of low molecular weight (LMW) hyaluronan as powder:

^* The water addition during the dough preparation corresponds to the water absorption in the Farinograph (-2.5%)

The use of LMW-HA leads to the increase in the bread volume and to the reduction in crumb strength (improved fresh-keeping).

The use of HMW-HA did not lead to a significant change in the bread properties, compared with the control.

Table 6: Baking experiments using different concentrations of high molecular weight (HMW) hyaluronan:

Table 7: Baking experiments using different concentrations of high and low molecular weight (HMW / LMW) hyaluronan

^* The water addition during the dough preparation corresponds to the water absorption in the Farinograph (-2.5%)

3. Digestibility

The glucose release upon digestion of DSC crumbs is shown in Figure 2. For this, bread was made from wheat flour with and without the addition of hyaluronan as powder.

After 1 and 3 days, the glucose release was measured.

Following the addition of 0.5% LMW-HA, a significantly lower release of glucose compared to the control was found, in particular after a storage time of 3 days. This includes not only the rapidly-digestible starch (given as "20 min" value), but also the slowly-digestible starch, denoted as "120 min" value.