METHOD OF PREPARING A CAKE USING PHOSPHOLIPASE

FIELD OF THE INVENTION

[0001] The present invention relates to a method of preparing a cake.

BACKGROUND OF THE INVENTION

[0002] Basic cake ingredients are usually flour, sugar, fats (from animal or vegetal origin), eggs and leaveners . Additional ingredients may be for example milk or milk fractions, flavorings or salt (in Pyler, E.J., 1988, Baking Science and Technology, Sosland Publishing, pp. 979-981). Eggs are commonly used in the preparation of various cakes. A certain amount of eggs is generally required to obtain a good cake quality, but eggs are an expensive ingredient, so it is desirable to reduce the amount of eggs and still achieve a satisfactory cake quality.

[0003] JP 63-258528A is directed to a method for producing a sponge cake by using egg liquid treated with phospholipase . [0004] JP 10-19187 IA is directed to a method for producing baked confectionery by treating a mixture with phospholipase before baking.

[0005] EP 0 426 211 Al is directed to a method of preparing a food product containing dried lysophospholipoprotein or dried lysophospholipoprotein containing material. In this invention egg yolk is treated during 4.5 hours at 54°C, the modified egg yolk is spray dried and the dried lysophospholipoprotein is added to dry cake mix, that after reconstitution with water and baking, results in a cake with an open and moist texture.

[0006] US 2003/0175383 Al is directed to a method of preparing a flour dough, said method comprising adding to the dough components an enzyme that under dough conditions is capable of hydrolysing a glycolipid and a phospholipid, wherein said enzyme is incapable, or substantially incapable, of hydrolyzing a triglyceride and/or a 1-monoglyceride, or a composition comprising said enzyme, and mixing the dough components to obtain a dough and to improve the strength and machinability of doughs and the volume, softness and crumb structure of bread and other baked products.

[0007] US 2003/0124647 Al is directed to a method of modifying whey protein in an aqueous solution by treating it with phospholipase . The modified whey protein shows to have improved foaming overrun and foam stability when whipped, as compared to whey protein preparation that is not treated with a phospholipase.

SUMNLARY OF THE INVENTION

[0008] The inventors confirmed that the volume and properties of a cake tend to deteriorate when the amount of eggs in the cake recipe is reduced.

[0009] They found that this deterioration can be counteracted by adding a phospholipase to the cake batter, as seen by an increased cake volume and improved cake properties, including the properties (of the fresh cake and also after storage), e.g. increased cohesiveness, increased springiness, and increased elasticity.

[0010] They found that the cake quality (as measured by these parameters) can be further improved, even up to the level of the original cake, by adding a non-egg protein together with the phospholipase.

[0011] Accordingly, in a first aspect the invention provides a method of preparing a cake, said method comprising preparing a cake batter by mixing cake batter ingredients,

said ingredients comprising non-phospholipase treated egg lecithin and phospholipase, and baking the cake batter to make the cake.

[0012] Accordingly, the invention provides a method of preparing a cake, comprising: a) preparing a cake batter comprising egg yolk lecithin, b) adding a phospholipase to the cake batter, and c) baking the cake batter to make the cake.

[0013] The method may further comprise adding a non-egg protein to the cake batter.

[0014] In another embodiment of the present invention the phospholipase is added to the mix of dry ingredient that is further mixed with other ingredients such as liquid eggs, oil, and water to prepare the batter.

DETAILED DESCRIPTION OF THE INVENTION

Phospholipase

[0015] The phospholipase is an enzyme that catalyzes the release of fatty acyl groups from a phospholipid. It may be a phospholipase A2 (PLA2, EC 3.1.1.4) or a phospholipase Al (EC 3.1.1.32) . It may or may not have other activities such as triacylglycerol lipase (EC 3.1.1.3) and/or galactolipase (EC 3.1.1.26) . [0016] The phospholipase may be a native enzyme derived from mammalian or microbial sources.

[0017] An example of a mammalian phospholipase is pancreatic PLA2, e.g. bovine or porcine PLA2 such as the commercial product Lecitase ^® 1OL (porcine PLA2, product of Novozymes A/S) . [0018] Microbial phospholipases may be derived from Fusarium, e.g. F. oxysporum phospholipase Al (WO 1998/026057), F. venenatum phospholipase Al (described in WO 2004/097012 as

a phospholipase A2 called FvPLA2), from Tuber, e.g. T. borchii phospholipase A2 (called TbPLA2, WO 2004/097012) .

[0019] The phospholipase may also be a lipolytic enzyme variant with phospholipase activity, e.g. as described in WO 2000/032758 or WO 2003/060112.

[0020] The phospholipase may be added in an amount of 500 - 20,000 units (LEU) per kg of batter, e.g. 1000-10,000 units

(LEU) per kg.

[0021] The phospholipase may also catalyze the release of fatty acyl groups from other lipids present in the batter, particularly wheat lipids. Thus, the phospholipase may have triacylglycerol lipase activity (EC 3.1.1.3) and/or galactolipase activity (EC 3.1.1.26).

Protein

[0022] Compared to a conventional cake recipe the amount of egg protein may be reduced and may be replaced by non-egg- protein. For example, compared to a conventional cake recipe, the amount of egg white protein may be reduced and may be replaced by non-egg protein.

[0023] Thus, the batter used in the invention may contain 0.5-3.0 % by weight of egg protein, and may contain 0.1-6 % (particularly 0.5-2 %) by weight of non-egg protein. For example, the batter used in the invention may contain 0.5-2.5 % by weight of egg white protein, and may contain 0.1-6 % (particularly 0.5-2 %) by weight of non-egg protein. [0024] The non-egg protein may particularly be a water- soluble, globular protein. The non-egg protein may particularly be partially or fully purified or isolated protein, such as, a water-soluble, globular protein. The non- egg protein may be denatured, and it may be one that partially unfolds to a rod-shaped or flexible molecule under the

interaction of lyso-lecithin formed by the action of the phospholipase on the egg yolk lecithin.

[0025] Protein sources with a good waterbinding, emulsifying and gelling properties in presence of lyso- phospholecithin are considered especially suitable

[0026] Examples of non-egg proteins are wheat proteins. Further examples of non-egg proteins are casein, whey protein, wheat gluten, legume protein (e.g. from soy bean, pea or lupine) . [0027] The non-egg protein may be subjected to a limited hydrolysis, e.g. enzymatic hydrolysis to 0-6 % hydrolysis. The enzymatic hydrolysis may be carried out with an amino-acid specific protease, e.g. one which is specific for Arg, Lys, GIu, Asp and/or Pro, such as the protease described in WO 91/13554.

[0028] The modification may include steps of shear treatment and acidic or alkaline pH, e.g. as described in WO2003/13266, increased temperature to denature partially or completely, protein deamidation, and separation steps including centrifugation, decanting and ultracentrifugation .

[0029] The protein (or hydrolyzed) protein may be enzymatically modified, e.g. with a cross-linking enzyme like transglutaminase or another protein modifying enzyme like protein-glutaminase . Furthermore the protein may be modified physically or chemically, e.g. through denaturation and deamidation .

Egg yolk lecithin

[0030] The cake batter comprises egg yolk lecithin, e.g. in the form of whole eggs, egg yolks, or egg powder.

[0031] The invention makes it possible to reduce the amount of egg material, e.g. to about half of a conventional cake. Thus, the batter may contain 0.3-1.5 % by weight of egg

lecithin or 5-25 % (particularly 7-20, or 8-15) by weight of whole eggs.

[0032] Advantageously, the batter may contain 0.1-1.5%, such as 0.1-1.2%, or 0.1-0.9%, or 0.2-1.5%, or 0.2-1.2%, or 0.2-0.9%, or 0.3-1.5 %, or 0.3-1.2%, or 0.3-0.9% by weight of egg lecithin or 5-25 % (particularly 7-20, or 8-15) by weight of whole eggs.

Other ingredients [0033] The cake batter may comprise other conventional ingredients, typically in the following amounts (in % by weight of the batter) :

- Flour (untreated, heat treated, chlorinated) : 15-30 %

- Starch (modified, native) : 0-10 % - Sugar: 15-25 %

- Emulsifier (mono and diglycerides of fatty acids, propylene glycol esters of fatty acids, lactic acid esters of mono and diglycerides of fatty acids, sodium stearoyl-2-lactylate) : 0.1-1 % - Baking powder (containing soda and acid or acidic salts) : 0.5-1 %

- Hydrocolloids (Locust bean gum, guar gum, tara gum, xanthan gum, carrageenan, acacia gum, cellulose, modified cellulose, pectin) : 0-1 % - Vegetable fat (ex. oil, margarine, shortening, fat paste, powdered fat) : 5-30 %

- Water: up to 100 %

[0034] Butter may advantageously replace part or all of the fat. [0035] An example of cake is a cake prepared with eggs - sugar - wheat flour - vegetable oil - starch - baking powder : sodium bicarbonate (E500ii) , sodium acid pyrophosphate (E450i) - emulsifier : mono and diglycerides of fatty acids (E471),

lactic acid esters of mono and diglycerides of fatty acids (E472b) , sodium stearoyl-2-lactylate (E481) - hydrocolloid : xanthan gum.

[0036] Another example of cake is a cake prepared with eggs - sugar - wheat flour -starch - margarine - baking powder : sodium bicarbonate (E500ii) , sodium acid pyrophosphate (E450i) - emulsifier : mono and diglycerides of fatty acids (E471) - propylene glycol esters of fatty acids (E477) lactic acid esters of mono and diglycerides of fatty acids (E472b) , sodium stearoyl-2-lactylate (E481) - hydrocolloid : carrageenan

[0037] A furher example of cake is a cake prepared with eggs - sugar - wheat flour - starch - margarine - baking powder : sodium bicarbonate (E500ii) , sodium acid pyrophosphate (E450i) - emulsifier : mono and diglycerides of fatty acids (E471) - propylene glycol esters of fatty acids (E477) - lactic acid esters of mono and diglycerides of fatty acids (E472b) - hydrocolloid : carrageenan

Assay methods

Phospholipase activity (LEU)

[0038] Lecithin is hydrolyzed under constant pH and temperature, and the phospholipase activity is determined as the rate of titrant (0.1N NaOH) consumption during neutralization of the liberated fatty acid. The substrate is soy lecithin (L-α-Phosphotidyl-Choline) , and the conditions are pH 8.00, 40.0 ⁰ C, reaction time 2 min. The method is further described in DK 99/00664 (Novo Nordisk A/S, Denmark) . The phospholipase from porcine pancreas has an activity of 510 LEU/mg and is taken as standard.

Texture Profile Analysis (TPA) for determination of cohesiveness and springiness

[0039] Two consecutive deformations of a cylindrical crumb sample (φ = 45 mm) performed with a cylindrical probe (φ = 100 mm) with a maximum deformation of 50% of the initial height of the product are performed at a deformation speed of 2mm/s and waiting time between consecutive deformations of 3s. Force is recorded as a function of time.

[0040] Cohesiveness is calculated as the ratio (expressed in percent) between the surface under the second deformation curve (downwards+upwards) and the surface under the first deformation curve (downwards+upwards) .

[0041] Springiness is calculated as the ratio between the height of the sample after the first deformation and 3 seconds waiting time and the initial height of the product.

Penetration test for determination of elasticity

[0042] Penetration of cake crumb with a cylindrical probe

(φ=25mm) until a total deformation of 25% of the initial height of the sample, at a deformation speed of 2 mm/s and keeping the target deformation constant during 20s. Force is registered as a function of time. Elasticity is the ratio (expressed in percent) between the force measured after 20s at constant deformation to the force applied to obtain the target deformation .

EXAMPLES

Example 1 : Effect of phospholipase

[0043] Cakes were made using a typical batter cake recipe. 15-25 % eggs, 20-30% flour, 0-10% starch, vegetable fat 15- 20%, 20-25% sugar, 0,1-1% emulsifier (mono and diglycerides of fatty acids, propylene glycol esters of fatty acids, lactic acid esters of mono and diglycerides of fatty acids, Sodium

Stearoyl-2-lactylate) , baking powder 0.8% (soda and SAPP (Sodium acid Pyrophosphate) ) , 0-1% hydrocolloids, 0-1% protein and water to 100% were mixed for 2 minutes at speed 2 (low) and 2 minutes at speed 5 (medium) in a Hobart mixer. [0044] Phospholipase was added directly to the dry mix, and finally eggs and oil and water were added to form the batter.

A total of 1.875 kg cake batter was prepared per trial. 30Og cake batter was weighed into aluminium pans.

[0045] The cakes were baked at a temperature of 180 ⁰ C for 45 minutes. 6 cakes with a total weight of 1.66 kg were made from each batter. Afterwards the cakes were cooled and packed in a plastic bag.

[0046] Textural properties were measured on day 1 and day

14 after baking using the method described above. Cohesiveness, springiness and elasticity as well as volume of the cakes were evaluated.

[0047] In the first example 1500 LEU/kg or 3750 LEU/kg was added to the batter where 50 % of the eggs (corresponding to

7.5-12.5 % by weight of the batter) were replaced by flour and water. A control was made with 100 % egg (corresponding to 15-

25 % by weight of the batter) ; the volume and textural properties were taken as 100%.

[0048] The following results show the effect of phospholipase on volume and texture of cake with 50% egg reduction and a comparison between microbial phospholipases and pancreatic phospholipase (Table 1).

Table 1

[0049] The results show that for 50% egg replaced by flour, the volume of the cake was only 90%, the cohesiveness on day 14 was only 70 %, and the elasticity on day 14 was 90% compared to the Control.

[0050] By the addition of TbPLA2, Lecitase 1OL, and FvPLA2 the volume of the 50% egg cakes was improved by 4-7%. 7% volume increase was achieved for 3750 LEU/kg batter Lecitase 1OL. [0051] The cohesiveness on day 14 was improved by 12-28%. Highest increase in cohesiveness was achieved by 3750LEU/kg batter Lecitase 1OL.

[0052] The elasticities of the resulting cakes were increased by 2-10% on day 14. Highest increase was measured for Lecitase 1OL (1500LEU/kg batter and 3750LEU/kg batter), FvPLA2 (3750LEU/kg batter) and TbPLA2 (1500LEU/kg batter) . [0053] Cake texture and cake volume were improved by all 3 phospholipases . Lecitase 1OL gave, with only 3% difference in volume, an elasticity and a cohesiveness on day 14 comparable to a Control cake with 100% egg.

Example 2 : Effect of combination of phospholipase and non-egg protein

[0054] Cakes were prepared as in Example 1, but with phospholipase (Lecitase 10L) and various non-egg proteins.

[0055] The following results show the effect of a combination of phospholipase and non-egg protein on volume and texture of cake prepared with 50% egg reduction (Table 2) . Provabis is a soy protein; the other proteins tested are all whey proteins.

[0056] The amount of protein (dry material) added in % by weight of the batter was 1.87-2.35 % for soy protein

(corresponding to all dry material of the replaced egg) and 0.935-1.175 % for the other proteins (corresponding to 50% of the dry material of the replaced egg) .

[0057] The non-egg proteins were commercial products from the following suppliers:

- Foamalac, Probake M, Carbelac 80 UHG: Carberry Group, Cork, Ireland - Hiprotal 45: Borculo Domo Ingredients, The Netherlands

- Lacprodan, Nutrilac BE-7602, Nutrilac BK-8310: Aria Foods Ingredients, Denmark

- Provabis: Cargill NV, Belgium

Hygel 8293, Hyfoama DSN: Kerry Bio-Science, The Netherlands

Table 2

[0058] The results (Table 2) show that by replacing 50 % of the eggs and adding a non-egg protein together and a phospholipase it was possible to reach the same cake volume and/or the same level of cohesiveness and/or elasticity after 14 days as the control.

[0059] 50% egg replaced by flour resulted in a volume loss of 15% compared to the Control. [0060] By the addition of Lecitase 1OL the volume was increased again by 9 %. With some of the non-egg proteins the volume of the 50% egg cake was improved to above the volume of the Control cake, while other proteins also showed an increase of the volume, but not up to the level of the control. [0061] Cohesiveness and elasticity were generally comparable or even above the values measured for the Control.

[0062] Springiness was improved by the non-egg proteins, but remained below the values measured for the Control on day 14.

[0063] Thus, the addition of non-egg protein together with a phospholipase can improve the volume, elasticity and cohesiveness of a 50% egg cake and make it comparable to the Control .

Example 3 : Effect of combination of phospholipase and non-egg protein

[0064] Cakes were prepared as in Example 1, but with addition of phospholipase and non-egg protein, alone or in combination (Table 3) .

Table 3

[0065] The effect of the non-egg protein is illustrated in the data where the addition of non-egg protein alone and in

combination with phospholipase is compared to 50% egg where egg has been replaced by flour and to 100% egg cakes

(=Control) .

[0066] It can be clearly seen that the addition of non-egg protein alone only gives slight improvement on the volume, while when combined with the Lecitase 1OL the volume is superior to the Control.

[0067] Here also the cohesiveness and the elasticity were comparable or above the values measured for the Control on day 14.

[0068] Springiness remains below the control on day 14.

Example 4 : Effect of combination of phospholipase and non-egg protein : wheat protein [0069] Cakes were prepared as in Example 1, but with phospholipase (Lecitase 10L) and various non-egg proteins, i.e. wheat proteins (Tables 4a to 4d) . A substantial amount of wheat proteins have been added such that the quantity of wheat protein is increased by at least 30% compared to the quantity originally present in the flour.

[0070] The following results show the effect of a combination of phospholipase and non-egg protein on volume and texture of cake prepared with 50% egg reduction. The amount of protein (dry material) added in % by weight of the batter was 0.9067 % and 1.813% (corresponding to respectively 50% and 100% of the dry material of the replaced egg) .

[0071] The non-egg proteins were commercial products from the following suppliers: - Prolite 100 and Prolite 200, ADM Speciality Food Ingredients, Keokuk, USA

- Meripro 420, Tate & LyIe Europe N. V., Belgium

- Gemtec 2170, Manildra Group, Auburn, Australia

HWG 2009, Loryma, Zwingenberg, Germany

Arise 5000, Midwest Grain Proteins, Atchison, Kansas, USA

Amygluten 110, Tate & LyIe Europe N. V., Belgium

Super Gluten 75 and Super Gluten 80, ADM

Glutastar EC75 and Glutastar EC80, Fiske Food Ingredients

Table 4a

Table 4b

Table 4c

5000

Arise

95 18 92 82 79 81 85 92 92 5000

Arise

95 18 3750 100 95 107 87 91 97 98 5000

Table 4d

[0072] The effect of the non-egg protein (s) is illustrated in the data presented on the tables 4a to 4d where the addition of non-egg protein (s) alone or in combination with phospholipase is compared to 50% egg formulations where egg has been replaced by flour and to 100% egg cakes (=Control) . [0073] Table 4a. When combining wheat protein and phospholipase (Lecitase 10L) in cake prepared with 50% less egg, volume of cake prepared with 100% egg can be completely recovered. When adding Prolite 100 or Prolite 200 in

combination with phospholipase to 50% egg formulations cohesiveness and elasticity of crumb 14 days after baking can be recovered or even increased and springiness is improved. [0074] Table 4b. Meripro 420, when added alone, has only a slight positive effect on volume of cake prepared with 50% less egg. Volume of cake prepared with 100% egg can be completely recovered by adding a combination of phospholipase and Meripro 420 to a 50% egg formulation. Cohesiveness is highly improved when adding both phospholipase and Meripro 420.

[0075] Table 4c. Wheat proteins tested, when added alone, have only a slight positive effect on volume. HWG2009 has a positive effect on volume but not on texture properties of cake prepared with 50% egg and phospholipase. Gemtec 2170 has no effect on volume of cake prepared with 50% egg and phospholipase but a positive effect on cohesiveness 14 days after baking and springiness 1 and 14 days after baking. Arise 5000 in combination with phospholipase has a positive effect on volume and cohesiveness of 50% egg cake. [0076] Table 4d. When adding wheat proteins : Meripro 420, Super Gluten 75, Super Gluten 80, Glutastar EC75 or Glutastar EC80 to a 50% egg cake recipe containing Lecitase 1OL, cohesiveness and resiliency measured 7 days after baking are significantly increased (between 7 and 9% ) when comparing these parameters measured 7 days after baking on 50% egg cake only containing Lecitase 1OL.

Example 5 : Effect of combination of phospholipase and non-egg proteins : sensorial analysis . [0077] Cakes were prepared as in Example 1, but without hydrocolloids and with phospholipase (Lecitase 10L) and two different non-egg proteins : Prolite 100 (ADM Speciality Food

Ingredients, Keokuk, USA) and Meripro 420 (Tate & LyIe Europe N. V., Aalst, Belgium).

[0078] Five different cake samples have been subjected to sensorial analysis by 29 subjects.

1. reference cake with 100% egg.

2. reference cake with 50% egg and replacement of egg by flour and water.

3. reference cake with 50% egg and replacement of egg by 3750 LEU lecitase 1OL / kg batter + 9g/kg batter Meripro 420 + 9g/kg batter of flour + water.

4. reference cake with 50% egg and replacement of egg by 3750 LEU lecitase 10 L / kg batter + 9g/kg batter Prolite 100 + 9g/kg batter of flour + water.

5. reference cake with 50% egg and replacement of egg by 3750 LEU lecitase 1OL /kg batter + 18g/kg batter Meripro 420.

[0079] Subjects have been asked to rank the different cakes according to their preference with the highest value for the highest appreciated cake and the lowest value for the less appreciated cake (Table 5) .

Table 5

[0080] The sum of rankings indicates that cake prepared with only 50% egg is less appreciated and that the four other types of cake are equally appreciated.

Example 6 : Effect of combination of phospholipase and non-egg protein: different types of cakes

[0081] Cakes were prepared with two different types of commercial dry mixes from Puratos (Brussels, Belgium) : Tegral Satin Cream Cake and Tegral Allegro Cake and with phospholipase (Lecitase 10L) and Meripro 420 (Tate & LyIe Europe N. V., Aalst, Belgium). The margarine added is Aristo Cake (Puratos, Brussels, Belgium) . The oil added is rapeseed oil. Batter is prepared and cakes are baked as described in example 1. The regular composition of the batters is given in table 6.

Table 6: Regular Batter composition

[0082] The following results show the effect of a combination of phospholipase and non-egg protein (s) on volume and texture of cake prepared with 50% egg reduction (Table 7) .

Table 7

[0083] Cake volume and texture were improved in recipe with 50% egg reduction by the use of a combination of non-egg protein (s) and phospholipase .