METHOD FOR PRODUCING MODIFIED PROTEIN-CONTAINING LIQUID FOOD

The present invention aims to provide a method for producing a protein-containing liquid food in which an unpleasant texture is improved, and the like. A method for producing a modified protein-containing liquid food, including treating a food ingredient containing a protein with phospholipase D. An enzyme preparation for modifying a protein-containing liquid food, which preparation contains phospholipase D. A method for modifying a protein-containing liquid food, including treating a food ingredient containing a protein with phospholipase D.

TECHNICAL FIELD

The present invention relates to a method for producing a modified protein-containing liquid food, an enzyme preparation for modifying a protein-containing liquid food, and a method for modifying a protein-containing liquid food.

BACKGROUND ART

Conventionally, various protein ingredients such as soy protein and milk protein have been widely used in liquid foods such as drinks, soup, and the like that contain protein, in order to reduce costs and impart nutritional value and richness of taste. In particular, due to the recent uncertainty about the supply of raw materials and the soaring prices of raw materials, companies have a strong demand for cost reduction, and studies are underway to maintain quality while reducing costs by adding inexpensive proteins. In addition, the demand for high-protein products is increasing due to the recent health boom, and products containing relatively large amounts of protein are desired. However, the addition of protein ingredients results in unpleasant textures such as “roughness” and “grittiness”, thus causing problems.

Therefore, a technique to improve the unpleasant texture derived from proteins in liquid foods has been desired.

Patent Literature 1 discloses a soy milk-containing liquid food or drink, which is characterized by mixing 0.5 to 30 parts of coconut milk relative to 100 parts of soy milk and a heat treatment at a temperature exceeding 100° C.

Cited document 2 discloses an oil-in-water emulsion to be added to a drink containing (a) caseinate and (b) a milk ingredient having a phospholipid content of 2 mass % or more in a milk-derived solid, in which (a)/(b) is 0.1 to 10 ((a)/(b) is a mass ratio of solids).

None of these documents describe the use of phospholipase D to improve the texture of protein-containing liquid foods.

CITATION LIST

Patent Literature

Patent Literature 1

Patent Literature 2

SUMMARY OF INVENTION

Technical Problem

The object of the present invention is to provide a method for producing a protein-containing liquid food (particularly a food containing a relatively large amount of protein ingredients) improved in an unpleasant texture, and the like.

Solution to Problem

The present inventors have conducted intensive studies in an attempt to solve the above-mentioned problems and found that a smooth texture can be imparted without imparting an unpleasant texture such as “roughness” or “grittiness” derived from protein, by adding a protein ingredient and phospholipase D (sometimes referred to as “PLD” in the present specification) in the production steps of liquid foods such as drinks and the like. Based on the finding, the present inventors conducted further studies and completed the present invention.

That is, the present invention provides the following.

Advantageous Effects of Invention

According to the present invention, a protein-containing liquid food, in which the protein-derived unpleasant texture is improved, can be provided.

According to the present invention, a liquid food in which the protein-derived unpleasant texture is suppressed can be provided, even when a relatively large amount of protein ingredient is added.

The present invention is applicable to a wide range of protein-containing liquid foods, such as plant-based foods.

DESCRIPTION OF EMBODIMENTS

1. Production Method of Modified Protein-Containing Liquid Food

The present invention relates to a method for producing a modified protein-containing liquid food.

The production method of the modified protein-containing liquid food of the present invention (hereinafter also to be simply referred to as the production method of the present invention) includes treating a food ingredient containing a protein with phospholipase D.

In the present invention, the protein-containing liquid foods include processed foods produced from food ingredients containing protein (hereinafter also to be simply referred to as “food ingredients”). Examples of the food ingredient containing protein include meats such as beef, pork, and chicken; fish such as Alaska pollock, hairtail, and threadfin bream; seafood (marine products) such as shellfish, shrimp, crab, octopus, and squid; grains such as rice and wheat; milk, egg, and proteins derived from plants or animals (for example, vegetable proteins such as soy protein, wheat protein, oat protein, pea protein, broad bean protein, mung bean protein, rice protein, chickpea protein, rapeseed protein, corn powder, Navy bean powder, almond protein, peanut powder, spirulina, soy milk, oat milk, and coconut milk; animal proteins such as egg white, egg white (powder), milk protein, skim milk powder, whey powder, casein or a salt thereof (for example, casein Na), cricket powder (Cricket, Big Cricket Protein), and Silkworm Powder); and the like.

In the present invention, the “protein-containing liquid food” refers to a food that contains protein and is in a liquid state (in other words, a state with flowability). In the present invention, the “protein-containing liquid food” only needs to be in a liquid state at the time of eating. For example, foods that are sold in a powdered or solid state and dissolved or dispersed in water or hot water by the purchaser when eaten (e.g., powdered drink, solid drink, powdered soup, solid soup) are also included in the “protein-containing liquid food” of the present invention.

Examples of the protein-containing liquid food include drinks (e.g., protein drinks, cafe au lait, plant-based milk (e.g., oat milk, almond milk, coconut milk, soy milk)), liquid seasonings (e.g., sauce, Tare sauce), liquid processed foods (e.g., soup, liquid diet), and plant-based foods in which the animal protein of these liquid foods is replaced with plant protein (plant-based (PB) drinks).

The embodiment of provision of the protein-containing liquid foods is not particularly limited. That is, the protein-containing liquid foods may be provided in any form, such as raw food, heated product, frozen product, aseptically packaged product, retort product, dried product, canned product, and the like.

In the present invention, even when a vegetable or animal-derived protein (e.g., vegetable protein such as soy protein or wheat protein; animal protein such as egg white, milk protein, casein or a salt thereof (e.g., casein Na), cricket powder, etc.) is contained in an amount of, for example, 0.1 wt % or more of the entire product (in the case of a food to be dissolved or dispersed in water or hot water when eaten, 0.1 wt % or more of the entire food after dissolution or dispersion), a product suppressed in the unpleasant texture derived from protein can be provided.

Phospholipase is an enzyme having the activity of hydrolyzing phospholipids.

In the present specification, the activity unit of phospholipase D is measured and defined as follows.

An enzyme solution (0.1 mL) is mixed with 0.9 mL of a substrate solution containing phosphatidylcholine, and reacted at 37° C. for 30 min. After discontinuation of the reaction, 50 μL of the reaction solution is added to 1 mL of color-developing solution containing choline oxidase, peroxidase, and the like, and reacted for 5 min. After discontinuation of the reaction, the amount of pigment produced from choline is measured. The amount of enzyme that liberates 1 μmol of choline per minute at 37° C. using phosphatidylcholine as a substrate is defined as 1 U (unit).

In the present invention, the amount of phospholipase D to be added is preferably 0.000000065 U or more, more preferably 0.00000065 U or more, further preferably 0.000065 or more, in terms of enzyme activity per 1 g of protein.

In the present invention, the amount of phospholipase D to be added is preferably 30,000 U or less, more preferably 15,000 U or less, and further preferably 6,494 or less, in terms of enzyme activity per 1 g of protein.

In the present invention, the amount of phospholipase D to be added is preferably 0.000000065 to 300000 U, more preferably 0.00000065 to 150000 U, further preferably 0.000065 to 6494 U, in terms of enzyme activity per 1 g of protein.

In the present invention, the amount of phospholipase D to be added is preferably 0.1 U or more, more preferably 1.2 to 10000.0 U, further preferably 12.0 to 5000.0 U, in terms of enzyme activity per 1 g of protein.

The action time (reaction time) of phospholipase D is not particularly limited as long as the enzyme can act on the phospholipid as a substrate substance. For example, it is 0 min or more, 1 min or more, 3 min or more, 5 min or more, 10 min or more, 20 min or more, or 30 min or more. For example, it is 168 hr or less, 72 hr or less, 48 hr or less, 24 hr or less, 12 hr or less, 6 hr or less, 3 hr or less, 2 hr or less, or 1 hr or less. A practical action time is preferably 0 to 148 hr, more preferably 30 min to 148 hr. The action temperature (reaction temperature) is also not particularly limited as long as the enzyme maintains its activity. A action at 0 to 60° C. is practically preferred. The enzyme reaction can be terminated by, for example, heating at 70 to 75° C. for 5 to 10 min.

In the production method of the present invention, it is preferable to further add, in addition to the above-mentioned phospholipase D, an enzyme that contributes to the formation of a cross-linked structure to the food ingredients and allow the enzyme to act.

In the present invention, an enzyme that contributes to the formation of a cross-linked structure is an enzyme that acts directly or indirectly on a protein and has the activity of forming a cross-linked structure in the protein.

In the present invention, examples of the enzyme that contributes to the formation of a cross-linked structure include ascorbic acid oxidase and glucose oxidase.

In the present invention, when ascorbic acid oxidase is used as the enzyme that contributes to the formation of a crosslinked structure, the food material to which the enzyme is added contains L-ASCORBIC ACID to be the substrate. The L-ASCORBIC ACID means ascorbic acid, ascorbate salt, or ascorbic acid with modified skeleton; examples include salts with alkali metal or alkaline earth metal (e.g., sodium ascorbate, calcium ascorbate, etc.), provitamin ascorbic acid 2-glucoside, ascorbic acid esters (e.g., ascorbyl palmitate, ascorbyl stearate, etc.), materials containing a lot of ascorbic acid, and the like. Among these, ascorbic acid and sodium ascorbate are preferred. Examples of the food material containing a lot of ascorbic acid include acerola powder and the like.

In the present invention, when ascorbic acid oxidase is used as the enzyme that contributes to the formation of a crosslinked structure, the amount of the L-ASCORBIC ACID in the food material to which the enzyme is added is, for example, 0.000000000001 to 50.0 weight, preferably 0.00000000001 to 30.0 wt %, more preferably 0.0000000001 to 10.0 wt %, further preferably 0.000000001 to 6.0 wt %, per gram of protein to which the enzyme is added.

In the present invention, when ascorbic acid oxidase is used as the enzyme that contributes to the formation of a crosslinked structure, the amount of the L-ASCORBIC ACID in the food material to which the enzyme is added is, for example, 0.1 to 99 wt %, preferably 1 to 95 wt %, more preferably 5 to 90 wt %, further preferably 10 to 80 wt %, calculated as ascorbic acid, relative to the agent of the present invention.

In the present invention, when glucose oxidase is used as the enzyme that contributes to the formation of a crosslinked structure, the food material to which the enzyme is added contains glucose to be the substrate.

In the present invention, when glucose oxidase is used as the enzyme that contributes to the formation of a crosslinked structure, the amount of the glucose in the food material to which the enzyme is added is 0.0000000001 to 10.0 weight, preferably 0.000000001 to 5.0 wt %, more preferably 0.00000001 to 1.0 wt %, further preferably 0.0000001 to 0.1 wt %, per gram of protein to which the enzyme is added.

In the present invention, when ascorbic acid oxidase is used as the enzyme that contributes to the formation of a crosslinked structure, the amount of the glucose in the food material to which the enzyme is added is, for example, 0.1 to 99 wt %, preferably 0.2 to 95 wt %, more preferably 0.5 to 90 wt %, further preferably 1 to 80 wt %, relative to the agent of the present invention.

When multiple enzymes are added, the order of addition may be any, and they may be added all at once or in sequence with a time lag. From the aspect of convenience, they are desirably added all at once.

When an enzyme that contributes to the formation of a cross-linked structure is further allowed to act on the food ingredients, the action time, action temperature, and method of terminating the enzyme reaction are the same as the action time, action temperature, and method of terminating the enzyme reaction for the above-mentioned phospholipase D.

In the production method of the present invention, the enzymes that act on the food ingredients include the following

The ascorbic acid oxidase (enzyme number EC1.10.3.3) used in the present invention is one of the ascorbic acid and aldaric acid metabolic enzymes, and is an oxidoreductase that catalyzes a chemical reaction that produces dehydroascorbic acid and water, using ascorbic acid and oxygen as substrates. Conventionally, ascorbic acid oxidase derived from Cucurbitaceae plants such as pumpkin, cucumber, and zucchini has been frequently used industrially. The origin of the ascorbic acid oxidase used in the present invention is not particularly limited as long as it has the above-mentioned activity, and may be derived from, for example, plant, microorganism, animal, or the like. In addition, the ascorbic acid oxidase to be used in the present invention may be a recombinant enzyme.

The method for producing the ascorbic acid oxidase to be used in the present invention is not particularly limited, and ascorbic acid oxidase produced by a method known per se or a method analogous thereto may be used. Commercially available ascorbic acid oxidase may also be used.

In the present invention, one type of ascorbic acid oxidase may be used alone, or two or more types may be used in combination.

In the present invention, as the activity unit of ascorbic acid oxidase, the amount of enzyme that oxidizes 1 μmol of ascorbic acid per minute under conditions of 30° C., pH 5.6 is defined as 1 U (unit).

Specifically, in the present invention, the activity of ascorbic acid oxidase is measured by the following procedures (1) to (3).

In the production method of the present invention, when ascorbic acid oxidase is used, the amount of the ascorbic acid oxidase to be added is, for example, 0.00000012 to 12000000000000 U, preferably 0.0000012 to 1200000000000 U, more preferably 0.000012 to 120000000000 U, further preferably 0.00012 to 12000000000 U, in terms of enzyme activity per 1 g of the content of the substrate of the enzyme (calculated as L-ascorbic acid).

In addition, in the production method of the present invention, when ascorbic acid oxidase is used, the amount of the ascorbic acid oxidase to be added is, for example, 0.5 to 500 U, preferably 1 to 350 U, more preferably 3 to 200 U, further preferably 5 to 100 U, in terms of enzyme activity per 1 g of the content of the substrate of the enzyme (calculated as L-ascorbic acid).

The glucose oxidase (EC1.1.3.4) to be used in the present invention is an enzyme that catalyzes a reaction in which glucose and oxygen are used as substrates to produce gluconolactone (gluconolactone is non-enzymatically hydrolyzed to gluconic acid) and hydrogen peroxide. The hydrogen peroxide produced by this reaction oxidizes the SH groups in proteins to promote the production of SS bond (disulfide bond) and form a cross-linked structure in protein. Glucose oxidases of various origins are known, including those derived from microorganisms such as Aspergillus oryzae and those derived from plants. Any of those glucose oxidases may be used, and the origin thereof is not limited. It may also be a recombinant enzyme. A specific example of glucose oxidase is the glucose oxidase derived from microorganism which is commercially available under the product name of “Sumizyme PGO” from Shin Nihon Chemical Co., Ltd.

As the activity unit of glucose oxidase in the present invention, the amount of enzyme that oxidizes 1 μmol of glucose per minute at 37° C. and pH=7.0 is defined as 1 U (unit).

For the activity of glucose oxidase in the present invention, the following method can be exemplified. Using glucose as a substrate, hydrogen peroxide is produced by the action of glucose oxidase in the presence of oxygen. The produced hydrogen peroxide is reacted with peroxidase in the presence of aminoantipyrine and phenol to produce quinoneimine dye. The produced quinoneimine dye is measured at a wavelength of 500 nm. Specifically, it is as follows. Glucose oxidase is stirred and dissolved in 0.1 mol/L phosphate buffer (adjusted to pH 7.0 with potassium dihydrogen phosphate and sodium hydroxide aqueous solution), and then diluted 50-fold with 0.1 mol/L phosphate buffer to obtain a GO solution. A phenol-containing buffer solution (2.0 mL) (obtained by mixing Milli-Q, 1.36 g of potassium dihydrogen phosphate, 3 mL of 5% phenol test solution, and 3 mL of 5% Triton X-100 solution and adjusted to pH 7.0, 100 mL with sodium hydroxide aqueous solution), 500 μL of 10% glucose solution, 500 μL of 0.01% peroxidase solution (using PO “amano” 3 (12500±250U)), and 100 μL of 0.4% 4-aminoantipyrine solution are added in this order to an analysis cell, mixed by inversion, and retained at 37±0.1° C. for 10 min. The GO solution (100 μL) is placed in the above-mentioned analysis cell, 11 points are automatically measured every 30 seconds for 5 min, and the GO activity value is measured from the increment (slope) between 120 seconds and 300 seconds. For the blank plot, the value measured by adding 0.1 mol/L phosphate buffer instead of the GO solution was used and subtracted from the value measured for the GO test plot. For oxidoreductases other than glucose oxidase, the amount of enzyme required to oxidize or reduce 1 μmol of substrate per minute is defined as 1 U (unit).

In the production method of the present invention, when glucose oxidase is used, the amount of the glucose oxidase to be added is, for example, 0.0000000022 to 215000000000 U, preferably 0.000000022 to 21500000000 U, more preferably 0.00000022 to 2150000000 U, further preferably 0.0000022 to 215000000 U, in terms of enzyme activity per 1 g of the substrate of the enzyme (glucose).

In the production method of the present invention, when glucose oxidase is used, the amount of the glucose oxidase to be added is, for example, 0.01 to 10000 U, preferably 0.1 to 5000 U, more preferably 0.5 to 3000 U, further preferably 1.0 to 2000 U, in terms of enzyme activity per 1 g of the substrate of the enzyme (glucose).

In the production method of the present invention, it is preferable to contain an auxiliary material selected from the following (A) to (N) in the food ingredients to which the enzyme is added. These auxiliary materials may be contained alone or in combination of two or more. In the production method of the present invention, by containing these auxiliary materials in the food ingredients to which the enzyme is added, a further improvement in smoothness (improvement of discomfort) can be expected compared to when they are not contained.

In the production method of the present invention, when an alkali salt is used as an auxiliary material, the amount of the alkali salt in the food material to which the enzyme is added is, for example, 0.0000000001 to 1.0 wt %, preferably 0.000000001 to 0.1 wt %, more preferably 0.00000001 to 0.06 wt %, further preferably 0.0000001 to 0.01 wt %, per gram of protein.

In the production method of the present invention, when a calcium salt or calcium oxide is used as an auxiliary material, the amount of the calcium salt or calcium oxide in the food material to which the enzyme is added is, for example, 0.0000000001 to 1.0 wt %, preferably 0.000000001 to 0.1 wt %, more preferably 0.00000001 to 0.06 wt %, further preferably 0.0000001 to 0.01 wt %, per gram of protein.

In the production method of the present invention, when a magnesium salt or magnesium oxide is used as an auxiliary material, the amount of the magnesium salt or magnesium oxide in the food material to which the enzyme is added is, for example, 0.0000000001 to 0.1 wt %, preferably 0.000000001 to 0.05 wt %, more preferably 0.00000001 to 0.01 wt %, further preferably 0.0000001 to 0.001 wt %, per gram of protein.

In the production method of the present invention, when a reducing agent is used as an auxiliary material, the amount of the reducing agent in the food material to which the enzyme is added is, for example, 0.000000000001 to 1.0 wt %, preferably 0.00000000001 to 0.5 wt %, more preferably 0.0000000001 to 0.1 wt %, further preferably 0.000000001 to 0.06 wt %, per gram of protein.

In the production method of the present invention, when a metal ion is used as an auxiliary material, the amount of the metal ion in the food material to which the enzyme is added is, for example, 0.0000000001 to 1.0 wt %, preferably 0.000000001 to 0.5 wt %, more preferably 0.00000001 to 0.1 wt %, further preferably 0.0000001 to 0.06 wt %, per gram of protein.

In the production method of the present invention, when a non-polar amino acid or non-polar amino acid salt is used as an auxiliary material, the amount of the non-polar amino acid or non-polar amino acid salt in the food material to which the enzyme is added is, for example, 0.000000000001 to 1.0 wt %, preferably 0.00000000001 to 0.5 wt %, more preferably 0.0000000001 to 0.1 wt %, further preferably 0.000000001 to 0.06 wt %, per gram of protein.

In the production method of the present invention, when an uncharged amino acid or uncharged amino acid salt is used as an auxiliary material, the amount of the uncharged amino acid or uncharged amino acid salt in the food material to which the enzyme is added is, for example, 0.00000000000001 to 1.0 wt %, preferably 0.0000000000001 to 0.1 wt %, more preferably 0.000000000001 to 0.06 wt %, further preferably 0.00000000001 to 0.01 wt %, per gram of protein.

In the production method of the present invention, when a basic amino acid or basic amino acid salt is used as an auxiliary material, the amount of the basic amino acid or basic amino acid salt in the food material to which the enzyme is added is, for example, 0.0000000001 to 0.1 wt %, preferably 0.000000001 to 0.05 wt %, more preferably 0.00000001 to 0.01 wt %, further preferably 0.0000001 to 0.001 wt %, per gram of protein.

In the production method of the present invention, when an acidic amino acid or acidic amino acid salt is used as an auxiliary material, the amount of the acidic amino acid or acidic amino acid salt in the food material to which the enzyme is added is, for example, 0.0000000001 to 0.1 wt %, preferably 0.000000001 to 0.05 wt %, more preferably 0.00000001 to 0.01 wt %, further preferably 0.0000001 to 0.001 wt %, per gram of protein.

The production method of the present invention can produce a protein-containing liquid food by using the same ingredients as those used for general protein-containing liquid foods and by a similar method, except that a treatment with the enzyme in the present invention is performed (when ascorbic acid oxidase or glucose oxidase is used, the ASCORBIC ACID or glucose to be the substrate is added to the ingredients) or preferably, the auxiliary materials described above are used. The enzyme in the present invention may be allowed to act on the food ingredients at any stage of the production step of the protein-containing liquid food. It may also be added and allowed to act during the step of producing protein ingredients. The enzyme in the present invention can be allowed to act on the food ingredients either as is, or by preparing an appropriate solution or the like and placing same in coexistence with the food ingredients. For example, the enzyme in the present invention may be added to the food ingredients, or the food ingredients may be immersed in a treatment solution containing the enzyme in the present invention. In the following, such operation to place the enzyme in the present invention in coexistence with the food ingredients is also to be collectively referred to as “addition” of the enzyme in the present invention.

The production method of the present invention can produce a modified protein-containing liquid food.

In the present specification, “modification” refers to imparting or enhancing a favorable texture (smoothness). In addition, “modification” also includes suppression of off-taste or off-flavor, or suppression of unpleasantness through modification.

The presence or absence of modification can be evaluated according to the sensory evaluation in the below-mentioned Experimental Examples.

2. Enzyme Preparation for Modifying Protein-Containing Liquid Food

The present invention also relates to an enzyme preparation for modifying protein-containing liquid food (hereinafter also to be simply referred to as the enzyme preparation of the present invention) containing phospholipase D.

In the enzyme preparation of the present invention, the definition and examples of protein-containing liquid food, examples of protein-containing food ingredients, examples of auxiliary materials, the amount of auxiliary materials in the food ingredients, and the definition, amount to be added, and method of addition (action time, action temperature, method of terminating the enzyme reaction) of phospholipase D are the same as the definition and examples of protein-containing liquid food, examples of protein-containing food ingredients, examples of auxiliary materials, the amount of auxiliary materials in the food ingredients, and the definition, amount to be added, and method of addition (action time, action temperature, method of terminating the enzyme reaction) of phospholipase D in the production method of the present invention.

In the enzyme preparation of the present invention, it is preferable to further contain, in addition to the above-mentioned phospholipase D, an enzyme that contributes to the formation of a cross-linked structure. In the enzyme preparation of the present invention, the definition, examples, amount to be added, and method of addition of the enzyme that contributes to the formation of a cross-linked structure are the same as the definition, examples, amount to be added, and method of addition of the enzyme that contributes to the formation of a cross-linked structure in the production method of the present invention.

The enzyme preparation of the present invention can be added to a food material containing protein (preferably a food material further containing the above-mentioned auxiliary material) and reacted according to the method and amount of addition of phospholipase D (or phospholipase D and an enzyme that contributes to the formation of a cross-linked structure), explained in the above-mentioned production method of the present invention, to produce a modified protein-containing liquid food.

3. Method for Modifying Protein-Containing Liquid Food

The present invention also relates to a method for modifying protein-containing liquid food (hereinafter also to be simply referred to as the modification method of the present invention), which includes treating a food ingredient containing a protein with phospholipase D.

In the modification method of the present invention, the definition and examples of protein-containing liquid food, examples of protein-containing food ingredients, examples of auxiliary materials, the amount of auxiliary materials in the food ingredients, and the definition, amount to be added, and method of addition (action time, action temperature, method of terminating the enzyme reaction) of phospholipase D are the same as the definition and examples of protein-containing liquid food, examples of protein-containing food ingredients, examples of auxiliary materials, the amount of auxiliary materials in the food ingredients, and the definition, amount to be added, and method of addition (action time, action temperature, method of terminating the enzyme reaction) of phospholipase D in the production method of the present invention.

The present invention is explained in more detail in the following by illustrating Examples and Experimental Examples; however, the present invention is not limited to these Examples and Experimental Examples.

Example

In the following Experimental Examples 1 to 4, the raw materials and equipment shown in Tables 1 and 2 were used.

raw materials used

raw material name
trade name
company name

wheat protein
Fumerit G
Nagata Group

casein Na
Casein Sodium LW
Nippon Shinyaku

Big Cricket Protein
Big Cricket Powder
TAKEO, Inc.

soy protein drink
SOY PROTEIN 100 
SAVAS

COCOA FLAVOR

Corporation

equipment used

equipment name
model
company name

vacuum 
A-300/16
Tokyo Food

packaging

Machinery

machine

[Experimental Example 1] Confirmation of Effect of Adding Phospholipase D in Soy Gel System

Soy gel samples 1-1 to 1-3 were prepared according to the sample preparation flow shown in FIG. 1, using the mixing recipe shown in Table 3. The prepared samples exhibit the properties of either a suspension or sol or gel.

The samples 1-1 to 1-3 obtained were subjected to a sensory evaluation of smoothness and off-taste or off-flavor by four expert panelists according to the following evaluation criteria. The results are shown in Table 4.

*2 PLD preparation: containing PLD (phospholipase D) 1.5 wt % ,dextrin 98.5 wt %; the number of PLD units per 1 g of PLD preparation is 820 U. The amount of PLD in sample 1-2 is 23.6 U when converted to enzyme activity for 1 g of protein in the sample.

flavor

From the results of Table 4, sample 1-2, with addition of a PLD preparation, was improved in smoothness compared to sample 1-1 (control).

[Experimental Example 2] Confirmation of Effect of Adding Phospholipase D in Various Protein Solution Gel Systems

Various protein gel samples 2-1 to 2-8, and various protein solution samples 2-9 to 2-12 were prepared according to the sample preparation flow shown in FIG. 2, using the mixing recipe shown in Table 5. The prepared samples exhibit the properties of either a suspension or sol or gel.

The samples 2-1 to 2-12 obtained were subjected to a sensory evaluation of smoothness by four expert panelists according to the following evaluation criteria. The results are shown in Table 6.

wheat protein

milk protein

casein Na

Cricket

Protein

Big Cricket

Protein

preparation

From the results of Table 6, sample 2-2, with a PLD preparation added to soybean protein gel, was improved in smoothness compared to sample 2-1 (control).

In addition, sample 2-4, with a PLD preparation added to wheat protein gel, was improved in smoothness compared to sample 2-3 (control).

In addition, sample 2-6, with a PLD preparation added to milk protein gel, was improved in smoothness compared to sample 2-5 (control).

In addition, sample 2-8, with a PLD preparation added to casein Na protein gel, was improved in smoothness compared to sample 2-7 (control).

In addition, sample 2-10, with a PLD preparation added to Cricket Protein solution, was improved in smoothness compared to sample 2-9 (control).

In addition, sample 2-12, with a PLD preparation added to Big Cricket Protein solution, was improved in smoothness compared to sample 2-11 (control).

[Experimental Example 3] Confirmation of Effect of Adding Phospholipase D in Soy Gel System

Soy gel samples 3-1 to 3-10 were prepared according to the sample preparation flow shown in FIG. 3, using the mixing recipe shown in Table 7. The prepared samples exhibit the properties of either a suspension or sol or gel.

The obtained samples 3-1 to 3-10 were subjected to a sensory evaluation of smoothness by four expert panelists according to the following evaluation criteria. The results are shown in Table 8.

raw
sample No.

preparation

From the results of Table 8, samples 3-2, 3-4, 3-6, 3-8, 3-10 obtained by adding PLD preparation to soybean protein gel were improved in smoothness compared to their respective controls, samples 3-1, 3-3, 3-5, 3-7, 3-9.

[Experimental Example 4] Confirmation of Effect of Adding Phospholipase D in Drink System

Drink samples 4-1 to 4-2 were prepared according to the sample preparation flow shown in FIG. 4, using the mixing recipe shown in Table 9.

The obtained drink samples 4-1 to 4-2 were subjected to a sensory evaluation of smoothness and off-taste or off-flavor by four expert panelists according to the following evaluation criteria. The results are shown in Table 10.

*1 soy protein drink: powdered drink to be dissolved in water when in use for drinking; soy protein content 75 wt %

*2 PLD preparation: containing PLD (phospholipase D) 1.5 wt %, dextrin 98.5 wt %; the number of PLD units per 1 g of PLD preparation is 820 U. The amount of PLD in sample 4-2 is 25.4 U when converted to enzyme activity for 1 g of protein in the sample.

flavor

From the results of Table 10, drink sample 4-2 produced by adding a PLD preparation to a soy protein drink was improved in smoothness and off-taste or off-flavor compared to sample 4-1 (control).

In the following Experimental Examples, unless otherwise specified, the raw materials and equipment shown in Tables 11 to 14 were used.

raw materials used

raw material name
trade name
company name

egg white (powder)
egg white powder
Taiyo Kagaku Co.,

wheat protein
Fumerit G
Nagata Group

Big Cricket Protein
Big Cricket Powder
TAKEO, Inc.

sodium chloride

ascorbic acid Na
Sodium L-ascorbate
Nippon Bulk Yakuhin

soy protein drink
SOY PROTEIN 100
SAVAS

COCOA FLAVOR

raw materials used

raw material name
trade name
company name

sodium carbonate
purified sodium
Daito Chemical Co.,

calcium chloride
calcium chloride
Tomita

calcium
calcium

calcium lactate
calcium lactate
FUSO CHEMICAL CO.,

GLUTAMATE
EUROPE

extract

yeast extract

iron-containing yeast
HIGH IRON
GROW COMPANY INC.

YEAST

yeast

raw materials used

raw material name
trade name
company name

hydrochloride

ammonium chloride
food additive
AKO KASEI CO.,

chloride MC

sodium alginate
Snow Algin H
Fuji Chemical

dietary fiber
Fuji FF
Fuji Nihon Seito

Corporation

OT
CORPORATION

CORPORATION

rapeseed protein
Puratein G
Merit Functional

Foods Corporation

corn powder
delicious corn
Hokkaido Knorr

whey powder
Morinaga whey
MORINAGA MILK

whole milk powder
Yotsuba Whole
Yotsuba Milk

Navy bean powder
Navy bean powder
Cargill Japan LLC

almond protein
Almond protein
Nissei Kyoeki

soy milk
Delicious
KIKKOMAN

Unsweetened
CORPORATION

Soy Milk

oat milk
alpro
DANONE JAPAN

Corporation

equipment used

equipment name
model
company name

vacuum packaging
A-300/16
Tokyo Food Machinery

machine

frozen cutter
FZ
Shonan Sangyo

Stephan cutter
UMC-5
Tokyo Food Machinery

small steamer
3-TYPE C
Yanagiya Machinery

food processor
RM-3200VD
FMI Corporation

[Experimental Example 5] Confirmation of Effect of Combined Use of Phospholipase D and Auxiliary Materials in Drink System

Drink samples 5-1 to 5-38 were prepared according to the sample preparation flow shown in FIG. 5, using the mixing recipes shown in Tables 15 to 19.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the following evaluation criteria. The results are shown in Table 20 to 24.

calcium chloride

yeast extract

yeast extract

(*1) soy protein drink: trade name SOY PROTEIN 100 COCOA FLAVOR, protein content 71.40%

(*2) The amount of PLD in samples 5-2, 5-4, 5-6, 5-8, 5-10 is 26.1 U when converted to enzyme activity for 1 g of protein in each sample.

calcium lactate

carbonic acid Na

(*1) soy protein drink: trade name SOY PROTEIN 100 COCOA FLAVOR, protein content 71.40%

(*2) The amount of PLD in samples 5-12, 5-14, 5-16, 5-18 is 26.1 U when converted to enzyme activity for 1 g of protein in each sample.

glycine

threonine

(*1) soy protein drink: trade name SOY PROTEIN 100 COCOA FLAVOR, protein content 71.40%

(*2) The amount of PLD in samples 5-20, 5-24, 5-26 is 26.1 U when converted to enzyme activity for 1 g of protein in each sample.

cysteine hydrochloride

alanine

cysteine

(*1) soy protein drink: trade name SOY PROTEIN 100 COCOA FLAVOR, protein content 71.40%

(*2) The amount of PLD in samples 5-28, 5-30, 5-32, 5-34 is 26.1 U when converted to enzyme activity for 1 g of protein in each sample.

glucose

(*1) soy protein drink: trade name SOY PROTEIN 100 COCOA FLAVOR, protein content 71.40%

(*2) The amount of PLD in samples 5-36, 5-38 is 26.1 U when converted to enzyme activity for 1 g of protein in each sample.

(*3) The amount of ASO in sample 5-36 is 1200 U when converted to enzyme activity for 1 g of Sodium L-ascorbate content (converted to L-ascorbic acid) in each sample.

(*4) The amount of GO in sample 5-38 is 2150 U, when converted to enzyme activity for 1 g of glucose in each sample.

From the results of Tables 20 to 23, the following was shown.

Sample 5-4, with the addition of PLD and calcium chloride thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-4 was improved in smoothness compared to sample 5-3 with the addition of PLD but without addition of calcium chloride.

Sample 5-6, with the addition of PLD and calcinated shell calcium thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-6 was improved in smoothness compared to sample 5-5 with the addition of PLD but without addition of calcinated shell calcium.

Sample 5-8, with the addition of PLD and glutathione-containing yeast extract thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-8 was improved in smoothness compared to sample 5-7 with the addition of PLD but without addition of glutathione-containing yeast extract.

Sample 5-10, with the addition of PLD and cysteine-containing yeast extract thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-10 was improved in smoothness compared to sample 5-9 with the addition of PLD but without addition of cysteine-containing yeast extract.

Sample 5-12, with the addition of PLD and cystine thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-12 was improved in smoothness compared to sample 5-11 with the addition of PLD but without addition of cystine.

Sample 5-14, with the addition of PLD and calcium lactate thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-14 was improved in smoothness compared to sample 5-13 with the addition of PLD but without addition of calcium lactate.

Sample 5-16, with the addition of PLD and carbonic acid Na thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-16 was improved in smoothness compared to sample 5-15 with the addition of PLD but without addition of carbonic acid Na.

Sample 5-18, with the addition of PLD and phosphoric acid 3Na thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-18 was improved in smoothness compared to sample 5-17 with the addition of PLD but without addition of phosphoric acid 3Na.

Sample 5-20, with the addition of PLD and iron-containing yeast thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-20 was improved in smoothness compared to sample 5-19 with the addition of PLD but without addition of iron-containing yeast.

Sample 5-24, with the addition of PLD and glycine thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-24 was improved in smoothness compared to sample 5-23 with the addition of PLD but without addition of glycine.

Sample 5-26, with the addition of PLD and threonine thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-26 was improved in smoothness compared to sample 5-25 with the addition of PLD but without addition of threonine.

Sample 5-28, with the addition of PLD and manganese-containing yeast thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-28 was improved in smoothness compared to sample 5-27 with the addition of PLD but without addition of manganese-containing yeast.

Sample 5-30, with the addition of PLD and cysteine hydrochloride thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-30 was improved in smoothness compared to sample 5-29 with the addition of PLD but without addition of cysteine hydrochloride.

Sample 5-32, with the addition of PLD and alanine thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-32 was improved in smoothness compared to sample 5-31 with the addition of PLD but without addition of alanine.

Sample 5-34, with the addition of PLD and cysteine thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-34 was improved in smoothness compared to sample 5-33 with the addition of PLD but without addition of cysteine.

From the results of Table 24, the following was shown. Sample 5-36, with the addition of PLD and L-ascorbic acid Na and ASO thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-36 was improved in smoothness compared to sample 5-2 (see Table 20) with the addition of PLD but without addition of L-ascorbic acid Na and ASO.

Sample 5-38, with the addition of PLD and glucose and GO thereto, was improved in smoothness compared to sample 5-1 (control). In addition, sample 5-38 was improved in smoothness compared to sample 5-2 (see Table 20) with the addition of PLD but without addition of glucose and GO.

[Experimental Example 6] Confirmation of Effect of Adding Phospholipase D to Plant-Based (PB) Drink

PB drink samples 6-1 to 6-9 were prepared according to the sample preparation flow shown in FIG. 6, using the mixing recipe shown in Table 25.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the following evaluation criteria. The results are shown in Table 26.

threonine

yeast extract

yeast

cysteine hydrochloride

glycine

alanine

cysteine

(*2) The amount of PLD in samples 6-2 to 6-9 is 20 U, when converted to enzyme activity for 1 g of protein in the sample.

From the results of Table 26, samples 6-2 to 6-9 obtained by adding PLD to PBP yogurt drinks were improved in smoothness compared to sample 6-1 (control). In addition, samples 6-3 to 6-9, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, cysteine hydrochloride, glycine, alanine, or cysteine) to PBP yogurt drinks, were improved in smoothness compared to sample 6-2 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

[Experimental Example 12] Confirmation of Effect of Adding Phospholipase D in Soy Gel System

Soy gel samples 12-2 to 12-15 were prepared according to the sample preparation flow shown in FIG. 7, using the mixing recipes shown in Tables 27-1, 27-2. In addition, egg white gel sample 12-1 was prepared according to the sample preparation flow shown in FIG. 7, using the mixing recipe shown in Table 27-1. The prepared samples exhibit the properties of either a suspension or sol or gel.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the following evaluation criteria, using sample 12-2 as a control. The results are shown in Tables 28-1, 28-2.

raw
soybean

material
protein (*1)

PLD activity (U) for 1

g of protein in sample

material
protein (*1)

g of protein in sample

From the results of Table 28-1, 28-2, samples 12-3 to sample 12-15 in which PLD was added such that the enzyme activity for 1 g of protein in the sample was in the range of 0.00000065 to 6494.3 U were improved in smoothness compared to sample 12-2 (control).

[Experimental Example 13] Confirmation of Effect of Combined Use of Phospholipase D and Auxiliary Materials in Soy Gel System

Classification of the fauxiliary materials used is shown in Table 29.

Each soy gel (egg white gel in sample 13-1) sample was prepared according to the sample preparation flow shown in FIG. 7, using the mixing recipes shown in Tables 29-1 to 29-41. The prepared samples exhibit the properties of either a suspension or sol or gel.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the same evaluation criteria as in Experimental Example 12, using sample 13-2 as a control. The results are shown in Tables 30-1 to 30-41.

auxiliary materials

auxiliary

classification
material No.
auxiliary material name

B: calcium salt or
B1
calcium chloride

C: magnesium salt or
C1
magnesium chloride

yeast extract

extract

amino acid salt
F3
alanine

acid or uncharged
G2
serine

amino acid salt
G3
glutamine

H: basic amino acid or
H1
arginine

basic amino acid salt
H2
histidine

or acidic amino acid
I2
sodium glutamate

(*3) The amount of PLD in sample 13-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-A1-2 to 13-A1-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-A2-2 to 13-A2-9 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-A3-2 to 13-A3-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in sample 13-A4-2 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-B1-2 to 13-B1-9 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-B2-2 to 13-B2-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-B3-3 to 13-B3-4 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-B4-2 to 13-B4-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-C1-2 to 13-C1-8 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-C2-2 to 13-C2-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

yeast extract

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-D1-2 to 13-D1-9 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

containing

yeast extract

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-D2-2 to 13-D2-10 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-E1-2 to 13-E1-9 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

yeast

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-E2-3 to 13-E2-6 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

yeast

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-E3-2 to 13-E3-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as *1 in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-F1-2 to 13-F1-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

protein

(*1) is the same as *1 in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-F2-2 to 13-F2-9 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-F3-2 to 13-F3-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-F4-2 to 13-F4-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as (*1) in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-F5-2 to 13-F5-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-F6-2 to 13-F6-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-F7-3 to 13-F7-4 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-F8-3 to 13-F8-4 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-F9-2 to 13-F9-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

raw
sample No.

protein

(*1) is the same as *1 in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-G1-2 to 13-G1-9 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-G2-2 to 13-G2-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-G3-2 to 13-G3-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-G4-3 to 13-G4-4 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-G5-3 to 13-G5-5 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as *1 in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-G6-3 to 13-G6-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-H1-2 to 13-H1-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-H2-2 to 13-H2-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-H3-2 to 13-H3-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(*1) is the same as *1 in the footnote of Table 29-1.

(*2) The amount of PLD in samples 13-I1-2 to 13-I1-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

*1 is the same as *1 in the footnote of Table 29-1.

*2 The amount of PLD in samples 13-12-3 to 13-12-4 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

From the results of Table 30-2, samples 13-A1-2 to 13-A1-7, with the addition of PLD and sodium carbonate, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-A1-2 to 13-A1-7 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of sodium carbonate.

From the results of Table 30-3, samples 13-A2-2 to 13-A2-9, with the addition of PLD and trisodium phosphate, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-A2-2 to 13-A2-9 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of trisodium phosphate.

From the results of Table 30-4, samples 13-A3-2 to 13-A3-3, with the addition of PLD and tripotassium phosphate, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-A3-2 to 13-A3-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of tripotassium phosphate.

From the results of Table 30-5, sample 13-A4-2, with the addition of PLD and trisodium citrate, was improved in smoothness compared to sample 13-2 (control). In addition, sample 13-A4-2 was improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of trisodium citrate.

From the results of Table 30-6, samples 13-B1-2 to 13-B1-9, with the addition of PLD and calcium chloride, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-B1-2 to 13-B1-9 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of calcium chloride.

From the results of Table 30-7, samples 13-B2-2 to 13-B2-7, with the addition of PLD and calcinated shell calcium, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-B2-2 to 13-B2-7 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of calcinated shell calcium.

From the results of Table 30-8, samples 13-B3-3 to 13-B3-4, with the addition of PLD and calcium lactate, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-B3-3 to 13-B3-4 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of calcium lactate.

From the results of Table 30-9, samples 13-B4-2 to 13-B4-3, with the addition of PLD and calcium carbonate, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-B4-2 to 13-B4-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of calcium carbonate.

From the results of Table 30-10, samples 13-C1-2 to 13-C1-8, with the addition of PLD and magnesium chloride, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-C1-2 to 13-C1-8 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of magnesium chloride.

From the results of Table 30-11, samples 13-C2-2 to 13-C2-3, with the addition of PLD and magnesium glutamate, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-C2-2 to 13-C2-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of magnesium glutamate.

From the results of Table 30-12, samples 13-D1-2 to 13-D1-9, with the addition of PLD and glutathione-containing yeast extract, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-D1-2 to 13-D1-9 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of glutathione-containing yeast extract.

From the results of Table 30-13, samples 13-D2-2 to 13-D2-10, with the addition of PLD and cysteine-containing yeast extract, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-D2-2 to 13-D2-10 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of cysteine-containing yeast extract.

From the results of Table 30-14, samples 13-E1-2 to 13-E1-9, with the addition of PLD and iron-containing yeast, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-E1-2 to 13-E1-9 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of iron-containing yeast.

From the results of Table 30-15, samples 13-E2-3 to 13-E2-6, with the addition of PLD and copper-containing yeast, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-E2-3 to 13-E2-6 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of copper-containing yeast.

From the results of Table 30-16, samples 13-E3-2 to 13-E3-7, with the addition of PLD and manganese-containing yeast, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-E3-2 to 13-E3-7 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of manganese-containing yeast.

From the results of Table 30-17, samples 13-F1-2 to 13-F1-7, with the addition of PLD and glycine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-F1-2 to 13-F1-7 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of glycine.

From the results of Table 30-18, samples 13-F2-2 to 13-F2-9, with the addition of PLD and cystine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-F2-2 to 13-F2-9 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of cystine.

From the results of Table 30-19, samples 13-F3-2 to 13-F3-3, with the addition of PLD and alanine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-F3-2 to 13-F3-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of alanine.

From the results of Table 30-20, samples 13-F4-2 to 13-F4-3, with the addition of PLD and valine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-F4-2 to 13-F4-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of valine.

From the results of Table 30-21, samples 13-F5-2 to 13-F5-3, with the addition of PLD and leucine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-F5-2 to 13-F5-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of leucine.

From the results of Table 30-22, samples 13-F6-2 to 13-F6-3, with the addition of PLD and isoleucine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-F6-2 to 13-F6-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of isoleucine.

From the results of Table 30-23, samples 13-F7-3 to 13-F7-4, with the addition of PLD and phenylalanine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-F7-3 to 13-F7-4 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of phenylalanine.

From the results of Table 30-24, samples 13-F8-3 to 13-F8-4, with the addition of PLD and proline, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-F8-3 to 13-F8-4 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of proline.

From the results of Table 30-25, samples 13-F9-2 to 13-F9-3, with the addition of PLD and methionine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-F9-2 to 13-F9-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of methionine.

From the results of Table 30-26, samples 13-G1-2 to 13-G1-9, with the addition of PLD and threonine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-G1-2 to 13-G1-9 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of threonine.

From the results of Table 30-27, samples 13-G2-2 to 13-G2-3, with the addition of PLD and serine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-G2-2 to 13-G2-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of serine.

From the results of Table 30-28, samples 13-G3-2 to 13-G3-3, with the addition of PLD and glutamine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-G3-2 to 13-G3-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of glutamine.

From the results of Table 30-29, samples 13-G4-3 to 13-G4-4, with the addition of PLD and tyrosine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-G4-3 to 13-G4-4 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of tyrosine.

From the results of Table 30-30, PLD added samples 13-G5-3 to 13-G5-5, with the addition of PLD and cysteine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-G5-3 to 13-G5-5 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of cysteine.

From the results of Table 30-31, samples 13-G6-3 to 13-G6-7, with the addition of PLD and cysteine hydrochloride, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-G6-3 to 13-G6-7 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of cysteine hydrochloride.

From the results of Table 30-32, samples 13-H1-2 to 13-H1-3, with the addition of PLD and arginine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-H1-2 to 13-H1-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of arginine.

From the results of Table 30-33, samples 13-H2-2 to 13-H2-3, with the addition of PLD and histidine, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-H2-2 to 13-H2-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of histidine.

From the results of Table 30-34, samples 13-H3-2 to 13-H3-3, with the addition of PLD and lysine hydrochloride, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-H3-2 to 13-H3-3 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of lysine hydrochloride.

From the results of Table 30-35, samples 13-I1-2 to 13-I1-7, with the addition of PLD and sodium aspartate, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-I1-2 to 13-I1-7 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of sodium aspartate.

From the results of Table 30-36, samples 13-I2-3 to 13-I2-4, with the addition of PLD and sodium glutamate, were improved in smoothness compared to sample 13-2 (control). In addition, samples 13-I2-3 to 13-I2-4 were improved in smoothness compared to sample 13-3 (see Table 30-1) with the addition of PLD but without addition of sodium glutamate.

[Experimental Example 14] Confirmation of (1) Effect of Combined Use of Phospholipase D and Ascorbic Acid Oxidase, and (2) Effect of Combined Use of Phospholipase D and Glucose Oxidase, in Soy Gel System

Each soy gel (egg white gel in sample 14-1) sample was prepared according to the sample preparation flow shown in FIG. 7, using the mixing recipes shown in Tables 31-1 to 31-9. The prepared samples exhibit the properties of either a suspension or sol or gel.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the same evaluation criteria as in Experimental Example 12, using sample 14-2 as a control. The results are shown in Tables 32-1 to 32-5.

raw
soybean protein (*1)

*3 The amount of PLD in sample 14-3 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(A) PLD × sodium ascorbate × ascorbic acid oxidase (study of the amount of sodium ascorbate)

ascorbic acid oxidase

ascorbic acid oxidase activity (U) for

1 g of Sodium L-ascorbate content

(converted to L-ascorbic acid) in sample

*2 The amount of PLD in samples 14A-6 to 14A-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

oxidase

activity (U) for 1 g of

Sodium L-ascorbate content

acid) in sample

*2 The amount of PLD in samples 14A-8 to 14A-14 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(B) PLD × sodium ascorbate × ascorbic acid oxidase (study of the amount of ascorbic acid oxidase)

ascorbic acid oxidase activity (U) for 1 g

of Sodium L-ascorbate content (converted

to L-ascorbic acid) in sample

*2 The amount of PLD in samples 14B-6 to 14B-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

the amount of ascorbic acid oxidase) <Mixing recipe>

oxidase

activity (U) for 1 g of

Sodium L-ascorbate content

acid) in sample

*2 The amount of PLD in samples 14B-8 to 14B-14 is 19.5 U when converted to enzyme activity for 1 g of

protein in the sample.

(C) PLD × glucose × glucose oxidase (study of the amount of glucose)

glucose oxidase

glucose oxidase activity (U)

for 1 g of glucose in sample

The amount of PLD in samples 14C-6 to 14C-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(C) PLD × glucose × glucose oxidase (study of the amount of glucose)

for 1 g of glucose in sample

*2 The amount of PLD in samples 14C-8 to 14C-11 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(D) PLD × glucose × glucose oxidase (study of the amount of glucose oxidase)

glucose

glucose oxidase activity (U)

for 1 g of glucose in sample

*2 The amount of PLD in samples 14D-6 to 14D-7 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

(D) PLD × glucose × glucose oxidase (study of the amount of glucose oxidase)

for 1 g of glucose in sample

*2 The amount of PLD in samples 14D-8 to 14D-14 is 19.5 U when converted to enzyme activity for 1 g of protein in the sample.

From the results of Tables 32-2, 32-3, samples 14A-6 to 14A-14 and samples 14B-6 to 14B-14, with the addition of PLD and ASO, were improved in smoothness compared to sample 14-2 (control). In addition, samples 14A-6 to 14A-14 and samples 14B-6 to 14B-14 were improved in smoothness compared to sample 14-3 (see Table 32-1) with the addition of PLD but without addition of ASO.

From the results of Tables 32-4, 32-5, samples 14C-6 to 14C-11 and samples 14D-6 to 14D-11, with the addition of PLD and GO, were improved in smoothness compared to sample 14-2 (control). In addition, samples 14C-6 to 14C-11 and samples 14D-6 to 14D-11 were improved in smoothness compared to sample 14-3 (see Table 32-1) with the addition of PLD but without addition of GO.

[Experimental Example 15] Confirmation of Effect of Adding Phospholipase D Alone and Effect of Combined Use of Phospholipase D and Auxiliary Materials, in Various Protein Systems

The trade name, company name, and protein content of various proteins used are shown in Table 33.

Each of various protein gel samples was prepared according to the sample preparation flow shown in FIG. 7, using the mixing recipes shown in Tables 33-1 to 33-29. The prepared samples exhibit the properties of either a suspension or sol or gel.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the following evaluation criteria. The results are shown in Tables 34-1 to 34-29.

raw material

protein

protein name
trade name
content
company name

CORPORATION

CORPORATION

CORPORATION

CORPORATION

CORPORATION

protein

Foods Corporation

egg white
egg white powder
83 wt %
Taiyo Kagaku Co.,

powder

powder

powder
Milk Powder

powder
Milk Powder

Navy bean
Navy bean powder
22 wt %
Cargill Japan LLC

powder

protein
powder

powder
Protein

Protein
Protein

Powder

material
pea protein

protein

mung bean

protein

PLD activity (U) for 1 g of

protein in sample

material
chickpea protein

egg white (powder)

PLD activity (U) for 1 g of

protein in sample

material
whey powder

powder

skim milk

powder

PLD activity (U) for 1 g of

protein in sample

powder

material
almond protein

Cricket

PLD activity (U) for 1 g of

protein in sample

raw
Big Cricket Protein
1.0
1.0

material
Silkworm Powder

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g

of protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g of

protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g of

protein in sample

material
Protein

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g of

protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g of

protein in sample

threonine

containing yeast

containing yeast

extract

alanine

cysteine

hydrochloride

cysteine

PLD activity (U) for 1 g of

protein in sample

material
pea protein

protein

mung bean

protein

PLD activity (U) for 1 g of

protein in sample

material
chickpea

protein

egg white

PLD activity (U) for 1 g of

protein in sample

material
whey powder

powder

skim milk powder

PLD activity (U) for 1 g of

protein in sample

material
powder

peanut powder

Cricket

PLD activity (U) for 1 g of

protein in sample

raw
Big Cricket Protein
20.0
20.0

material
Silkworm Powder

PLD activity (U) for 1 g of

protein in sample

From the results of Tables 34-1 to 34-29, the following was shown.

Sample 15A-2 (PLD activity value 297.4 U/protein 1 g) obtained by adding PLD to oat protein was improved in smoothness compared to sample 15A-1 (control).

Sample 15A-4 (PLD activity value 29.7 U/protein 1 g) obtained by adding PLD to oat protein was improved in smoothness compared to sample 15A-3 (control).

Sample 15A-12 (PLD activity value 14.9 U/protein 1 g) obtained by adding PLD to oat protein was improved in smoothness compared to sample 15A-11 (control).

Sample 15B-2 (PLD activity value 215.4 U/protein 1 g) obtained by adding PLD to pea protein was improved in smoothness compared to sample 15B-1 (control).

Sample 15B-4 (PLD activity value 21.5 U/protein 1 g) obtained by adding PLD to pea protein was improved in smoothness compared to sample 15B-3 (control).

Sample 15B-12 (PLD activity value 10.8 U/protein 1 g) obtained by adding PLD to pea protein was improved in smoothness compared to sample 15B-11 (control).

Sample 15C-2 (PLD activity value 201.8 U/protein 1 g) obtained by adding PLD to broad bean protein was improved in smoothness compared to sample 15C-1 (control).

Sample 15C-4 (PLD activity value 20.2 U/protein 1 g) obtained by adding PLD to broad bean protein was improved in smoothness compared to sample 15C-3 (control).

Sample 15C-12 (PLD activity value 10.1 U/protein 1 g) obtained by adding PLD to broad bean protein was improved in smoothness compared to sample 15C-11 (control).

Sample 15D-2 (PLD activity value 226.0 U/protein 1 g) obtained by adding PLD to mung bean protein was improved in smoothness compared to sample 15D-1 (control).

Sample 15D-4 (PLD activity value 22.6 U/protein 1 g) obtained by adding PLD to mung bean protein was improved in smoothness compared to sample 15D-3 (control).

Sample 15D-12 (PLD activity value 11.3 U/protein 1 g) obtained by adding PLD to mung bean protein was improved in smoothness compared to sample 15D-11 (control).

Sample 15E-2 (PLD activity value 226.0 U/protein 1 g) obtained by adding PLD to rice protein was improved in smoothness compared to sample 15E-1 (control).

Sample 15E-4 (PLD activity value 22.6 U/protein 1 g) obtained by adding PLD to rice protein was improved in smoothness compared to sample 15E-3 (control).

Sample 15E-12 (PLD activity value 11.3 U/protein 1 g) obtained by adding PLD to rice protein was improved in smoothness compared to sample 15E-11 (control).

Sample 15F-2 (PLD activity value 269.0 U/protein 1 g) obtained by adding PLD to chickpea protein was improved in smoothness compared to sample 15F-1 (control).

Sample 15F-4 (PLD activity value 26.9 U/protein 1 g) obtained by adding PLD to chickpea protein was improved in smoothness compared to sample 15F-3 (control).

Sample 15F-12 (PLD activity value 13.5 U/protein 1 g) obtained by adding PLD to chickpea protein was improved in smoothness compared to sample 15F-11 (control).

Sample 15G-4 (PLD activity value 18.8 U/protein 1 g) obtained by adding PLD to rapeseed protein was improved in smoothness compared to sample 15G-3 (control).

Sample 15G-12 (PLD activity value 9.4 U/protein 1 g) obtained by adding PLD to rapeseed protein was improved in smoothness compared to sample 15G-11 (control).

Sample 15H-2 (PLD activity value 204.2 U/protein 1 g) obtained by adding PLD to egg white was improved in smoothness compared to sample 15H-1 (control).

Sample 15H-4 (PLD activity value 20.4 U/protein 1 g) obtained by adding PLD to egg white was improved in smoothness compared to sample 15H-3 (control).

Sample 15H-12 (PLD activity value 10.2 U/protein 1 g) obtained by adding PLD to egg white was improved in smoothness compared to sample 15H-11 (control).

Sample 151-2 (PLD activity value 1461.2 U/protein 1 g) obtained by adding PLD to corn protein was improved in smoothness compared to sample 151-1 (control).

Sample 151-4 (PLD activity value 146.1 U/protein 1 g) obtained by adding PLD to corn protein was improved in smoothness compared to sample 151-3 (control).

Sample 151-12 (PLD activity value 73.1 U/protein 1 g) obtained by adding PLD to corn protein was improved in smoothness compared to sample 151-11 (control).

Sample 15J-2 (PLD activity value 1356.0 U/protein 1 g) obtained by adding PLD to whey protein was improved in smoothness compared to sample 15J-1 (control).

Sample 15J-4 (PLD activity value 135.6 U/protein 1 g) obtained by adding PLD to whey protein was improved in smoothness compared to sample 15J-3 (control).

Sample 15J-12 (PLD activity value 67.8 U/protein 1 g) obtained by adding PLD to whey protein was improved in smoothness compared to sample 15J-11 (control).

Sample 15K-2 (PLD activity value 625.5 U/protein 1 g) obtained by adding PLD to whole milk protein powder was improved in smoothness compared to sample 15K-1 (control).

Sample 15K-4 (PLD activity value 62.5 U/protein 1 g) obtained by adding PLD to whole milk protein powder was improved in smoothness compared to sample 15K-3 (control). Sample 15K-12 (PLD activity value 31.3 U/protein 1 g) obtained by adding PLD to whole milk protein powder was improved in smoothness compared to sample 15K-11 (control).

Sample 15L-2 (PLD activity value 476.1 U/protein 1 g) obtained by adding PLD to skim milk protein was improved in smoothness compared to sample 15L-1 (control).

Sample 15L-4 (PLD activity value 47.6 U/protein 1 g) obtained by adding PLD to skim milk protein was improved in smoothness compared to sample 15L-3 (control).

Sample 15L-12 (PLD activity value 23.8 U/protein 1 g) obtained by adding PLD to skim milk protein was improved in smoothness compared to sample 15L-11 (control).

Sample 15M-2 (PLD activity value 767.0 U/protein 1 g) obtained by adding PLD to Navy bean protein was improved in smoothness compared to sample 15M-1 (control).

Sample 15M-4 (PLD activity value 76.7 U/protein 1 g) obtained by adding PLD to Navy bean protein was improved in smoothness compared to sample 15M-3 (control).

Sample 15M-12 (PLD activity value 38.3 U/protein 1 g) obtained by adding PLD to Navy bean protein was improved in smoothness compared to sample 15M-11 (control).

Sample 15N-2 (PLD activity value 385.2 U/protein 1 g) obtained by adding PLD to almond protein was improved in smoothness compared to sample 15N-1 (control).

Sample 15N-4 (PLD activity value 38.5 U/protein 1 g) obtained by adding PLD to almond protein was improved in smoothness compared to sample 15N-3 (control).

Sample 15N-12 (PLD activity value 19.3 U/protein 1 g) obtained by adding PLD to almond protein was improved in smoothness compared to sample 15N-11 (control).

Sample 150-2 (PLD activity value 368.5 U/protein 1 g) obtained by adding PLD to peanut protein was improved in smoothness compared to sample 150-1 (control).

Sample 150-4 (PLD activity value 36.8 U/protein 1 g) obtained by adding PLD to peanut protein was improved in smoothness compared to sample 150-3 (control).

Sample 150-12 (PLD activity value 18.4 U/protein 1 g) obtained by adding PLD to peanut protein was improved in smoothness compared to sample 150-11 (control).

Sample 15P-2 (PLD activity value 327.9 U/protein 1 g) obtained by adding PLD to Cricket protein was improved in smoothness compared to sample 15P-1 (control).

Sample 15P-4 (PLD activity value 32.8 U/protein 1 g) obtained by adding PLD to Cricket protein was improved in smoothness compared to sample 15P-3 (control).

Sample 15P-12 (PLD activity value 16.4 U/protein 1 g) obtained by adding PLD to Cricket protein was improved in smoothness compared to sample 15P-11 (control).

Sample 15Q-2 (PLD activity value 306.5 U/protein 1 g) obtained by adding PLD to Big Cricket protein was improved in smoothness compared to sample 150-1 (control).

Sample 15Q-4 (PLD activity value 30.7 U/protein 1 g) obtained by adding PLD to Big Cricket protein was improved in smoothness compared to sample 150-3 (control).

Sample 15Q-12 (PLD activity value 15.3 U/protein 1 g) obtained by adding PLD to Big Cricket protein was improved in smoothness compared to sample 150-11 (control).

Sample 15R-2 (PLD activity value 309.9 U/protein 1 g) obtained by adding PLD to Silkworm protein was improved in smoothness compared to sample 15R-1 (control).

Sample 15R-4 (PLD activity value 31.0 U/protein 1 g) obtained by adding PLD to Silkworm protein was improved in smoothness compared to sample 15R-3 (control).

Sample 15R-12 (PLD activity value 15.5 U/protein 1 g) obtained by adding PLD to Silkworm protein was improved in smoothness compared to sample 15R-11 (control).

Sample 15S-2 (PLD activity value 274.3 U/protein 1 g) obtained by adding PLD to spirulina protein was improved in smoothness compared to sample 15S-1 (control).

Sample 15S-4 (PLD activity value 27.4 U/protein 1 g) obtained by adding PLD to spirulina protein was improved in smoothness compared to sample 15S-3 (control).

Sample 15S-12 (PLD activity value 13.7 U/protein 1 g) obtained by adding PLD to spirulina protein was improved in smoothness compared to sample 15S-11 (control).

From the results of Table 34-6, samples 15A-5 to 15A-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to oat protein, were improved in smoothness compared to sample 15A-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-7, samples 15B-5 to 15B-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to pea protein, were improved in smoothness compared to sample 15B-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-8, samples 15C-5 to 15C-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to broad bean protein, were improved in smoothness compared to sample 15C-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-9, samples 15D-5 to 15D-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to mung bean protein, were improved in smoothness compared to sample 15D-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-10, samples 15E-5 to 15E-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to rice protein, were improved in smoothness compared to sample 15E-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-11, samples 15F-5 to 15F-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to chickpea protein, were improved in smoothness compared to sample 15F-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-12, samples 15G-5 to 15G-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to rapeseed protein, were improved in smoothness compared to sample 15G-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-13, samples 15H-5 to 15H-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to egg white, were improved in smoothness compared to sample 15H-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-14, samples 151-5 to 151-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to corn protein, were improved in smoothness compared to sample 151-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-15, samples 15J-5 to 15J-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to whey protein, were improved in smoothness compared to sample 15J-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-16, samples 15K-5 to 15K-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to whole milk protein powder, were improved in smoothness compared to sample 15K-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-17, samples 15L-5 to 15L-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to skim milk protein, were improved in smoothness compared to sample 15L-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-18, samples 15M-5 to 15M-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to Navy bean protein, were improved in smoothness compared to sample 15M-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-19, samples 15N-5 to 15N-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to almond protein, were improved in smoothness compared to sample 15N-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-20, samples 150-5 to 150-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to peanut protein, were improved in smoothness compared to sample 150-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-21, samples 15P-5 to 15P-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to Cricket protein, were improved in smoothness compared to sample 15P-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-22, samples 150-5 to 150-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to Big Cricket protein, were improved in smoothness compared to sample 15Q-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-23, samples 15R-5 to 15R-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to Silkworm protein, were improved in smoothness compared to sample 15R-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 34-24, samples 15S-5 to 15S-10, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to spirulina protein, were improved in smoothness compared to sample 15S-4 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

[Experimental Example 16] Confirmation of Effect of Adding Phospholipase D Alone and Effect of Combined Use of Phospholipase D and Auxiliary Materials, in Various Protein Systems

The trade name, company name, and protein content of various proteins used are shown in Table 35.

Each protein gel sample was prepared according to the sample preparation flow shown in FIG. 7, using the mixing recipes shown in Tables 35-1 to 35-3. The prepared samples exhibit the properties of either a suspension or sol or gel.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the following evaluation criteria. The results are shown in Tables 36-1 to 36-3.

protein

name of milk
trade name
content
company name

Corporation

Soy Milk

coconut milk
coconut milk
2
wt %
YOUKI FOOD

threonine

yeast

yeast extract

alanine

cysteine hydrochloride

cysteine

(*1) The amount of PLD in samples 16A-2 to 16A-8 is 20.4 U when converted to enzyme activity for 1 g of protein in the sample.

threonine

yeast

yeast extract

alanine

cysteine hydrochloride

cysteine

(*1) The amount of PLD in samples 16B-2 to 16B-8 is 847.5 U when converted to enzyme activity for 1 g of protein in the sample.

threonine

yeast

yeast extract

alanine

cysteine hydrochloride

cysteine

(*1) The amount of PLD in samples 16C-2 to 16C-8 is 89.2 U when converted to enzyme activity for 1 g of protein in the sample.

From the results of Table 36-1, samples 16A-2 to 16A-8, obtained by adding PLD to soy milk, were improved in smoothness compared to sample 16A-1 (control). In addition, 16A-3 to 16A-8, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to soy milk, were improved in smoothness compared to sample 16A-2 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 36-2, samples 16B-2 to 16B-8, obtained by adding PLD to oat milk, were improved in smoothness compared to sample 16B-1 (control). In addition, 16B-3 to 16B-8, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to oat milk, were improved in smoothness compared to sample 16B-2 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 36-3, samples 16C-2 to 16C-8, obtained by adding PLD to coconut milk, were improved in smoothness compared to sample 16C-1 (control). In addition, 16C-3 to 16C-8, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to coconut milk, were improved in smoothness compared to sample 16C-2 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

[Experimental Example 17] Confirmation of Effect of Adding Phospholipase D Alone and Effect of Combined Use of Phospholipase D and Auxiliary Materials, in Various Protein Systems

The trade name, company name, and protein content of various proteins used are shown in Table 37.

Each protein gel sample was prepared according to the sample preparation flow shown in FIG. 8, using the mixing recipes shown in Tables 37-1 to 37-3.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the following evaluation criteria. The results are shown in Tables 38-1 to 38-3.

protein

meat name
product name
content
company name

beef belly
beef belly
21 wt %
Tokyo Packer

pork arm
pork arm
22 wt %
Tokyo Packer

breast
breast

threonine

yeast extract

alanine

cysteine hydrochloride

cysteine

(*1) The amount of PLD in samples 17A-2 to 17A-8 is 9.6 U when converted to enzyme activity for 1 g of protein in the sample.

threonine

yeast extract

alanine

cysteine hydrochloride

cysteine

(*1) The amount of PLD in samples 17B-2 to 17B-8 is 9.3 U when converted to enzyme activity for 1 g of protein in the sample.

threonine

yeast extract

alanine

cysteine hydrochloride

cysteine

(*1) The amount of PLD in samples 17C-2 to 17C-8 is 8.8 U when converted to enzyme activity for 1 g of protein in the sample.

From the results of Table 38-1, samples 17A-2 to 17A-8, obtained by adding PLD to beef belly (lean only), were improved in smoothness compared to sample 17A-1 (control). In addition, 17A-3 to 17A-8, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to beef belly (lean only), were improved in smoothness compared to sample 17A-2 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 38-2, samples 17B-2 to 17B-8, obtained by adding PLD to pork arm (lean only), were improved in smoothness compared to sample 17B-1 (control). In addition, 17B-3 to 17B-8, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to pork arm (lean only), were improved in smoothness compared to sample 17B-2 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 38-3, samples 17C-2 to 17C-8, obtained by adding PLD to chicken breast, were improved in smoothness compared to sample 17C-1 (control). In addition, 17C-3 to 17C-8, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, alanine, cysteine hydrochloride, or cysteine) to chicken breast, were improved in smoothness compared to sample 17C-2 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

[Experimental Example 18] Confirmation of Effect of Adding Phospholipase D Alone and Effect of Combined Use of Phospholipase D and Auxiliary Materials, in Various Protein Systems

Protein gel samples 18A-1 to 18A-6 were prepared according to the sample preparation flow shown in FIG. 9 (no sitting step (generally a step of leaving a meat paste at a low temperature of around 10 to 40° C. for a certain period of time)), using the mixing recipe shown in Table 39-1.

In addition, protein gel samples 18B-1 to 18B-6 were prepared according to the sample preparation flow shown in FIG. 9 (with sitting step), using the mixing recipe shown in Table 39-2.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the following evaluation criteria. The results are shown in Tables 40-1, 40-2.

threonine

extract

cysteine hydrochloride

(*2) The amount of PLD in samples 18A-2 to 18A-6 is 14.7 U when converted to enzyme activity for 1 g of protein in the sample.

threonine

extract

cysteine hydrochloride

(*2) The amount of PLD in samples 18B-2 to 18B-6 is 14.7 U when converted to enzyme activity for 1 g of protein in the sample.

From the results of Table 40-1, samples 18A-2 to 18A-6, obtained by adding PLD to hairtail C, were improved in smoothness compared to sample 18A-1 (control). In addition, 18A-3 to 18A-6, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, or cysteine hydrochloride) to hairtail C, were improved in smoothness compared to sample 18A-2 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

From the results of Table 40-2, samples 18B-2 to 17B-6, obtained by adding PLD to hairtail C, were improved in smoothness compared to sample 18B-1 (control). In addition, samples 18B-3 to 18B-6, obtained by adding PLD and an auxiliary material (threonine, manganese-containing yeast, glutathione-containing yeast extract, or cysteine hydrochloride) to hairtail C, were improved in smoothness compared to sample 18B-2 with the addition of PLD but without addition of the above-mentioned auxiliary materials.

[Experimental Example 19] Comparative Verification of Phospholipase D and Existing Materials in Soy Gel System

Each protein gel sample was prepared according to the sample preparation flow shown in FIG. 10, using the mixing recipes shown in Tables 41-1, 41-2. The prepared samples exhibit the properties of either a suspension or sol or gel.

The obtained each sample was subjected to a sensory evaluation of smoothness and off-taste or off-flavor by three expert panelists according to the following evaluation criteria. The results are shown in Tables 42-1, 42-2.

material
protein (*1)

of protein in sample

protein in sample

PLD activity (U) for 1 g of protein

in sample

viscosity
and not

and not
evaluable

From the results of Tables 42-1, 42-2, samples 19-12 to 19-14, obtained by adding PLD to soy gel, were improved in smoothness compared to sample 19-1 (control). In addition, samples using high amounts of lecithin, PLA1, or PLA2 (samples 19-4, 19-5, 19-8, 19-11) had an off-taste or off-flavor, whereas samples 19-12 to 19-14 containing PLD were shown to be superior in that they had high scores of smoothness and no off-taste or off-flavor.

[Experimental Example 20] Confirmation of Effect of Adding Phospholipase D in Soy Gel System Produced by One-Step Heating

Soy gel samples 20-1 to 20-5 were prepared according to the sample preparation flow shown in FIG. 7, using the mixing recipe shown in Table 43. The prepared samples exhibit the properties of either a suspension or sol or gel.

The obtained each sample was subjected to a sensory evaluation of smoothness by three expert panelists according to the same evaluation criteria as in Experimental Example 12, using sample 21-2 as a control. The results are shown in Table 44.

raw
soybean protein (*1)

PLD activity (U) for 1 g of protein in

sample

From the results of Table 44, samples 20-3 to 20-5 obtained by adding PLD to soy gel were improved in smoothness compared to sample 20-2 (control).

INDUSTRIAL APPLICABILITY

According to the present invention, a protein-containing liquid food, in which the protein-derived unpleasant texture is improved, can be provided.

This application is based on patent application No. 2023-022041 filed in Japan, the contents of which are encompassed in full herein.