Enzyme preparation

Abstract

To provide a protein deamidase agent having improved thermal stability.SOLUTION: A guanidinium compound and polyamine can improve the thermal stability of a protein deamidase.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an enzyme preparation of protein deamidase having excellent thermostability. [Background technology]

[0002] Protein deamidase is used as a food additive enzyme, and improves physical functions such as solubility, emulsifying properties, foaming properties, and gelling properties of proteins and protein materials in foods by utilizing its action of converting glutamine residues in food proteins to glutamic acid residues. For this reason, protein deamidase is expected to be used in various applications including foods and beverages (Patent Document 1). In particular, with the recent increase in health consciousness, the consumption of plant proteins has accelerated, and protein deamidase has high potential value in the processing of plant proteins. On the other hand, protein deamidase has a problem of poor thermal stability (Non-Patent Document 1). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 2000-50887 A [Non-patent literature]

[0004] [Non-Patent Document 1] Appl Microbiol Biotechnol. 104; 187-199. Summary of the Invention [Problem to be solved by the invention]

[0005] In view of the possibility that the range of application of protein deamidase in protein processing will continue to expand in the future, it is desired to be able to utilize protein deamidase in a state in which its thermal stability is improved.

[0006] Therefore, an object of the present invention is to provide an enzyme preparation of protein deamidase having improved thermostability. [Means for solving the problem]

[0007] As a result of intensive research, the present inventors have found that the thermal stability of protein deamidase is improved by causing the protein deamidase to coexist with a guanidium compound and / or a polyamine. The present invention has been completed based on this finding and through further research.

[0008] That is, the present invention provides the following aspects. Item 1. An enzyme preparation comprising (A) a protein deamidase and (B) a guanidinium compound and / or a polyamine. Item 2. The enzyme preparation according to Item 1, wherein the guanidinium compound is arginine and / or argininamide. Item 3. The enzyme preparation according to Item 1 or 2, wherein the polyamine is spermidine. Item 4. The enzyme preparation according to any one of Items 1 to 3, wherein the content of the component (B) per 1 U of the component (A) is 0.5 μmol or more. Item 5. The enzyme preparation according to any one of Items 1 to 4, wherein the content of the component (B) is 0.05 M or more. Item 6. The enzyme preparation according to any one of Items 1 to 5, wherein the protein deamidase is protein glutaminase. Item 7. The enzyme preparation according to any one of Items 1 to 6, which is a liquid preparation. Item 8. A method for deamidating a protein, comprising the step of treating a protein (P) with a protein deamidase (A) in the presence of a guanidium compound and / or a polyamine (B) to proceed with a deamidation reaction. Item 9. A method for producing a deamidated protein, comprising a step of proceeding with a deamidation reaction of component (P) in a reaction mixture comprising a protein (P), (A) a protein deamidase, and (B) a guanidium compound and / or a polyamine. Item 10. The production method according to Item 9, wherein the reaction mixture is prepared by mixing the component (P) with an enzyme preparation containing the component (A) and the component (B). Item 11. The production method according to Item 9, wherein the reaction mixture is prepared by mixing a mixture containing the component (P) and the component (B) with the component (A). Item 12. The method according to any one of Items 9 to 11, wherein the deamidation reaction is carried out at a temperature higher than the optimum temperature for the component (A). Item 13. The method according to any one of Items 9 to 12, wherein the component (P) is a vegetable protein and / or an animal protein. Item 14. A method for producing a food, beverage, or pharmaceutical, comprising a step of proceeding with a deamidation reaction of the component (P) in a reaction mixture containing a food, beverage, or pharmaceutical raw material containing a protein (P), (A) a protein deamidase, and (B) a guanidium compound and / or a polyamine. Effect of the Invention

[0009] According to the present invention, there are provided an enzyme preparation of protein deamidase having improved thermostability, a method for deamidating protein in a state in which protein deamidase has high thermostability, and a method for producing a deamidated protein utilizing the same. [Brief description of the drawings]

[0010] [Figure 1] These are the results of verifying the effect of various amino acids on PG thermal stability. [Diagram 2] These are the results of a verification of the effect of different concentrations of arginine on the thermal stability of PG. [Diagram 3] These are the results of verifying the effects of arginine, argininamide, and spermidine on the thermal stability of PG. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] 1. Enzyme preparations The enzyme preparation of the present invention is characterized by comprising (A) a protein deamidase (hereinafter also referred to as "component (A)") and (B) a guanidium compound and / or a polyamine (hereinafter also referred to as "component (B)"). The enzyme preparation of the present invention will be described in detail below.

[0012] 1-1.(A) Protein deamidase The enzyme preparation of the present invention contains protein deamidase as component (A). Protein deamidase is an enzyme that deamidates amide groups of glutamine residues and asparagine residues in proteins without cleaving peptide bonds or crosslinking proteins.

[0013] Examples of protein deamidating enzymes used in the enzyme preparation of the present invention include the following. (1) Enzymes that deamidate glutamine residues in proteins, converting them to glutamic acid (e.g., protein glutaminase). (2) Enzymes that deamidate asparagine residues in proteins, converting them to aspartic acid (e.g., protein asparaginase). (3) Enzymes that deiminate arginine residues in proteins and convert them to citrulline (e.g., arginine deiminase, protein arginine deiminase, peptidylarginine deiminase).

[0014] In general, when glutamine and asparagine residues in a protein are deamidated to generate carboxyl groups, the negative charge of the protein increases, resulting in a decrease in the isoelectric point and an increase in hydration. Furthermore, the increase in electrostatic repulsion leads to a decrease in the interaction between proteins, i.e., a decrease in association. These changes greatly increase the solubility and water dispersibility of the protein. In addition, the increase in the negative charge of the protein unfolds the protein, changes its higher-order structure, and exposes the hydrophobic region buried inside the molecule to the molecular surface. Therefore, the deamidated protein has amphiphilic properties and becomes an ideal surfactant, greatly improving the emulsifying power, emulsion stability, foaming property, and foam stability of the protein. In this way, the deamidation of a protein improves various functional properties of the protein, and the uses of the protein increase dramatically. In addition, when the arginine residues in a protein are deiminated, the hydrophobicity of the protein is increased and the higher-order structure of the protein is changed.

[0015] More specific examples of protein deamidation enzymes include those of the genus Chryseobacterium, Flavobacterium, Empedobacter, Sphingobacterium, Aureobacterium, and the like, which are disclosed in JP2000-50887A, JP2001-218590A, and WO2006 / 075772A1. Examples of commercially available products of protein deamidase derived from the genus Myroides or the genus Luteimicrobium, Agromyces, Microbacterium, or Leifsonia disclosed in WO2015 / 133590, and protein glutaminase derived from the genus Chryseobacterium. In the enzyme preparation of the present invention, one of these protein deamidases may be used alone as component (A), or a combination of two or more of them may be used.

[0016] Among these protein deamidating enzymes, from the viewpoint of further enhancing the effect of improving thermal stability by the coexistence with component (B), preferably, protein deamidating enzyme derived from the genus Chryseobacterium is used, more preferably, protein glutaminase derived from the genus Chryseobacterium is used, and even more preferably, protein glutaminase derived from the species Chryseobacterium proteolyticum is used.

[0017] Protein deamidase can be prepared from a culture solution of a microorganism from which the above-mentioned protein deamidase is derived. Specific preparation methods include a method of recovering protein deamidase from a culture solution or cells of the above-mentioned microorganism. For example, when a protein deamidase-secreting microorganism is used, the cells can be recovered from the culture solution in advance by filtration, centrifugation, or the like as necessary, and the enzyme can be separated and / or purified. When a protein deamidase-nonsecreting microorganism is used, the cells can be recovered from the culture solution in advance by pressure treatment, ultrasonic treatment, or the like to expose the enzyme, and the enzyme can be separated and / or purified. As a method for separating and / or purifying the enzyme, a known protein separation and / or purification method can be used without any particular limitation, and examples thereof include centrifugation, UF concentration, salting out, various chromatography methods using ion exchange resins, or the like.

[0018] The content of component (A) in the enzyme preparation of the present invention is not particularly limited, and may be appropriately set to an extent that the enzyme activity of component (A) can be utilized when the enzyme preparation of the present invention is used, and may be, for example, 1 U / mL or more, preferably 10 U / mL or more, more preferably 25 U / mL or more, even more preferably 50 U / mL or more, and particularly preferably 90 U / mL or more in terms of protein deamidation enzyme activity. The upper limit of the content of component (A) may be, for example, 10,000 U / mL or less, preferably 1,000 U / mL or less, more preferably 500 U / mL or less, even more preferably 200 U / mL or less, and particularly preferably 110 U / mL or less.

[0019] Regarding the activity of protein deamidase, the amount of enzyme that liberates 1 μmol of ammonia per minute using benzyloxycarbonyl-L-glutaminylglycine (Z-Gln-Gly) as a substrate is defined as 1 unit (1 U).

[0020] 1-2.(B) Guanidinium compounds and / or polyamines The enzyme preparation of the present invention contains a guanidinium compound and / or a polyamine as component (B), which improves the thermal stability of component (A).

[0021] Guanidinium compounds have the guanidino group NH 2 There is no particular limitation as long as it is a compound having -C(=NH)-NH-. Examples of the guanidium compound used in the present invention include guanidine, arginine, and arginine amide. These guanidium compounds may be used alone or in combination of two or more. Among these guanidium compounds, from the viewpoint of further enhancing the effect of improving the thermal stability of the (A) component, arginine and arginine amide are preferred, and arginine amide is more preferred.

[0022] The polyamine is not particularly limited as long as it is a straight-chain aliphatic hydrocarbon having three or more primary amino groups bonded thereto. From the viewpoint of further enhancing the effect of improving the thermal stability of the component (A), a polyamine is preferably used as the component (B). Examples of the polyamine used in the present invention include spermine and spermidine. These polyamines may be used alone or in combination of two or more kinds. Among these polyamines, spermidine is preferred from the viewpoint of further enhancing the effect of improving the thermal stability of the component (A).

[0023] The ratio of the content of the (A) component to the (B) component in the enzyme preparation of the present invention may be appropriately set according to the desired degree of the thermal stability improving effect of the (A) component. The content of the (B) component per 1 U of the (A) component is, for example, 0.5 μmol or more, and from the viewpoint of further enhancing the thermal stability improving effect of the (A) component, it is preferably 0.8 μmol or more, more preferably 4 μmol or more, and even more preferably 9 μmol or more. The upper limit of the content of the (B) component per 1 U of the (A) component is not particularly limited, but it is, for example, 50 μmol or less, preferably 30 μmol or less, and more preferably 15 μmol or less.

[0024] The specific content of the (B) component in the enzyme preparation of the present invention is determined according to the content of the (A) component and the above ratio, but may be, for example, 0.05 M or more, and from the viewpoint of further enhancing the effect of improving the thermal stability of the (A) component, may be preferably 0.08 M or more, more preferably 0.4 M or more, and even more preferably 0.9 M or more. The upper limit of the content of the (B) component in the enzyme preparation of the present invention is not particularly limited, but may be, for example, 5 M or less, preferably 3 M or less, and more preferably 1.5 M or less.

[0025] 1-3.Other ingredients The enzyme preparation of the present invention may contain other ingredients besides the above-mentioned components (A) and (B), such as other enzymes, additives, solvents, and culture residue components.

[0026] Examples of other enzymes include amylase (α-amylase, β-amylase, glucoamylase), glucosidase (α-glucosidase, β-glucosidase), galactosidase (α-galactosidase, β-galactosidase), protease (acid protease, neutral protease, alkaline protease), peptidase (leucine peptidase, aminopeptidase), lipase, esterase, cellulase, phosphatase (acid phosphatase, alkaline phosphatase), nuclease, deaminase, oxidase, dehydrogenase, glutaminase, pectinase, catalase, dextranase, transglutaminase, pullulanase, etc. These other enzymes may be contained alone or in combination of multiple types.

[0027] Examples of additives include excipients, buffers, suspending agents, stabilizers, preservatives, antiseptics, and pH adjusters. Examples of excipients include starch, dextrin, maltose, trehalose, lactose, D-glucose, sorbitol, D-mannitol, sucrose, and glycerol. Examples of buffers include phosphates, citrates, and acetates. Examples of stabilizers include propylene glycol and ascorbic acid. Examples of preservatives include phenol, benzalkonium chloride, benzyl alcohol, chlorobutanol, and methylparaben. Examples of preservatives include ethanol, benzalkonium chloride, paraoxybenzoic acid, and chlorobutanol. These additives may be contained alone or in combination. Examples of solvents include water. The content of these additives and solvents may be appropriately set according to the types of the components and / or the properties of the enzyme preparation.

[0028] Examples of the culture residue components include components contained in the medium used for culturing the bacteria that produce component (A) or other enzymes that are blended as necessary, impurity proteins and bacterial components generated by the culture, etc. These culture residue components may be contained alone or in combination of multiple types.

[0029] 1-4.Properties The properties of the enzyme preparation of the present invention are not particularly limited, and examples thereof include liquid and solid (powder, granules, etc.). Since the enzyme preparation of the present invention has an excellent effect of improving the thermal stability of the component (A), even if the enzyme preparation is in a liquid form, which is inherently unstable, it can effectively improve the thermal stability. From this perspective, a preferred example of the properties of the enzyme preparation of the present invention is a liquid form (liquid preparation).

[0030] When the enzyme preparation of the present invention is a liquid preparation, the pH (25° C.) of the liquid preparation is, for example, 6.5 to 7.5, preferably 6.8 to 7.2, and more preferably 6.9 to 7.1.

[0031] 1-5.Applications The enzyme preparation of the present invention can be used in any application that utilizes protein deamidase activity. In other words, the enzyme preparation of the present invention can be used in any protein modification application that utilizes changes in protein properties brought about by deamidation of γ-amide and β-amide groups in glutamine and asparagine residues of a protein to generate carboxyl groups. Specific examples of protein modification include increased protein solubility, increased water dispersibility, improved emulsifying power, and improved emulsion stability.

[0032] Preferred applications of the enzyme preparation of the present invention include applications in the food and pharmaceutical fields, and more preferred applications include applications in the food field. Specifically, applications include applications for improving the solubility, dispersibility, emulsification, etc. of animal proteins and / or vegetable proteins in a weakly acidic condition environment that is the pH range of normal foods (for example, applications for producing acidic beverages such as coffee whitener, juice, dressings, mayonnaise, and creams); increasing the solubility and dispersibility of poorly soluble vegetable proteins (for example, applications for producing tempura flour using wheat gluten); applications for modifying dough in bread and confectionery (for example, applications for producing crackers, biscuits, cookies, pizza or pie crusts); and applications for removing or reducing allergens in allergenic proteins in foods. (e.g., in the production of foods for patients with wheat allergies); uses for reducing protein mineral sensitivity, increasing the soluble mineral content in liquids containing proteins and minerals, and increasing the absorbability of minerals into the human body (e.g., in the production of high mineral (e.g., calcium) content beverages and mineral (e.g., calcium) absorption enhancers); uses for reducing bitterness, improving the protein hydrolysis rate of proteases, and / or enhancing the glutamic acid content (e.g., in the production of amino acid-based seasonings (hydrolyzed animal protein (HAP), hydrolyzed vegetable protein (HVP)), miso and soy sauce), etc.

[0033] 2. Method for deamidating proteins and method for producing deamidated proteins As described above, a guanidium compound and / or a polyamine can improve the thermal stability of protein deamidase, and therefore, a guanidium compound and / or a polyamine can be allowed to coexist in an enzyme reaction system using protein deamidase.

[0034] Thus, the present invention also provides a method for deamidating a protein, comprising the step of treating a (P) protein (hereinafter also referred to as a "(P) component") with (A) protein deamidase in the presence of (B) a guanidium compound and / or a polyamine, thereby proceeding with a deamidation reaction.

[0035] The protein deamidation method of the present invention can also be used for producing a deamidated protein. Accordingly, the present invention also provides a method for producing a deamidated protein, comprising the step of proceeding a deamidation reaction of the component (P) in a reaction mixture containing a protein (P), (A) protein deamidase, and (B) a guanidium compound and / or a polyamine. Furthermore, the method for producing a deamidated protein of the present invention can be used for producing a food, beverage, or medicine. Accordingly, the present invention also provides a method for producing a food, beverage, or medicine, comprising the step of proceeding a deamidation reaction of the component (P) in a reaction mixture containing a food, beverage, or medicine raw material containing the protein (P), (A) protein deamidase, and (B) a guanidium compound and / or a polyamine.

[0036] Hereinafter, the protein deamidation method and the method for producing a deamidated protein of the present invention (including the method for producing foods, beverages, and pharmaceuticals) are collectively referred to as the "method, etc. of the present invention."

[0037] 2-1.(P) Protein In the method and production method of the present invention, the protein as component (P) to be subjected to protein deamidation may be either a vegetable protein or an animal protein. In the method and production method of the present invention, either a vegetable protein or an animal protein may be used as component (P), or both may be used in combination.

[0038] 2-2. Procedure In the methods etc. of the present invention, it is sufficient that component (B) coexists in an enzymatic reaction system in which component (A) acts on component (P). Therefore, the order in which these components are mixed is not particularly limited as long as a reaction mixture containing component (P), component (A), and component (B) is prepared when the reaction mixture is subjected to conditions in which the deamidation reaction of component (P) proceeds.

[0039] For example, in one example of the method of the present invention, the enzyme preparation described above in "1. Enzyme Preparation" is used, and the enzyme preparation is mixed with component (P) to prepare the reaction mixture. In this case, since component (A) contained in the enzyme preparation has excellent thermal stability, the activity of component (A) can be efficiently enjoyed. In addition, the activity of component (A) can be stably exerted even under the heating conditions applied in the step of progressing the deamidation reaction.

[0040] In another example of the method of the present invention, the reaction mixture can be prepared by mixing the component (A) with a mixture containing the component (P) and the component (B). In this case, the activity of the component (A) can be stably exerted under the heating conditions applied in the step of proceeding with the deamidation reaction.

[0041] 2-3. Amount of each ingredient used In the method of the present invention, the amount of the (A) component used relative to the (P) component can be the same as that used in known methods without any particular restrictions. The ratio of the amounts of the (A) component and the (B) component used may be appropriately set according to the degree of the desired effect of improving the thermal stability of the (A) component. The amount of the (B) component used per 1 U of the (A) component is, for example, 0.5 μmol or more, and from the viewpoint of further enhancing the effect of improving the thermal stability of the (A) component, is preferably 0.8 μmol or more, more preferably 4 μmol or more, and even more preferably 9 μmol or more. The upper limit of the amount of the (B) component used per 1 U of the (A) component is not particularly limited, but may be, for example, 50 μmol or less, preferably 30 μmol or less, and more preferably 15 μmol or less.

[0042] 2-4.Reaction conditions The reaction conditions (temperature, time, pH, etc.) in the step of proceeding with the deamidation reaction are not particularly limited as long as a deamidated protein can be obtained. Optimal treatment conditions may be determined through preliminary experiments.

[0043] Among these reaction conditions, the temperature conditions in the step of promoting the deamidation reaction may be appropriately set according to the optimum temperature of component (A). Since component (A) used in the method of the present invention has excellent thermal stability, the temperature conditions may be higher than the optimum temperature of component (A). In this case, the temperature conditions may be acceptable even if the temperature is such that component (A) is inactivated in the absence of component (B). Specifically, the temperature conditions may be, for example, 5 to 15°C, preferably 8 to 12°C, higher than the optimum temperature of component (A).

[0044] 2-5. Other processes The method etc. of the present invention may include other steps other than the step of proceeding with the deamidation reaction. The other steps are not particularly limited as long as they are suitable for the applications described in "1-5. Applications", and examples thereof include an enzyme inactivation step, a cooling step, a filtration step, a cooking step, etc. These other steps may be performed individually or in combination of two or more steps. EXAMPLES

[0045] The present invention will be specifically described below by way of examples, but the present invention should not be construed as being limited to the following examples.

[0046] [Protein deamidation enzyme] Protein Glutaminase “Aman” o"500; Chryseobacterium proteolyticum derived protease Protein glutaminase (manufactured by Amano Enzyme Inc.) was used. This protein glutaminase has an optimum temperature of 50 to 60° C., and is inactivated by treatment at 70° C. for 1 hour. Hereinafter, this protein deamidase is also referred to as "PG."

[0047] The protein deamidase activity was measured by the following method. 0.1 mL of the sample solution containing protein deamidase was added to 1 mL of 0.2 M phosphate buffer (pH 6.5) containing 30 mM Z-Gln-Gly, and the mixture was left at 37° C. for 10 minutes, after which 1 mL of 0.4 M TCA solution was added to stop the reaction. As a blank, 1 mL of 0.4 M TCA solution was added to 1 mL of 0.2 M phosphate buffer (pH 6.5) containing 30 mM Z-Gln-Gly, and 0.1 mL of the sample solution containing protein deamidase was further added and left at 37° C. for 10 minutes.

[0048] The amount of ammonia generated in the reaction solution was measured using an Ammonia Test Wako (FUJIFILM Wako Pure Chemical Industries, Ltd.) The ammonia concentration in the reaction solution was determined from a calibration curve showing the relationship between ammonia concentration and absorbance (630 nm) prepared using an ammonia standard solution (ammonium chloride).

[0049] The activity of protein deamidase was calculated from the following formula, where 1 unit (1U) is the amount of enzyme that produces 1 μmol of ammonia per minute, where the reaction solution volume is 2.1, the enzyme solution volume is 0.1, Df is the dilution ratio of the enzyme solution, and 17.03 is the molecular weight of ammonia.

[0050]

number

[0051] [Thermal stability measurement method] An enzyme solution containing component (A) (PG) at a final concentration of 100 U / mL and containing or not containing component (B) was prepared. The pH of the prepared enzyme solution at 25°C was 7.0. The enzyme solution was subjected to heat treatment at 70°C for a predetermined time, and the degree of thermal aggregation of the enzyme was evaluated by measuring the absorbance at 413 nm. A higher absorbance indicates a higher degree of the enzyme forming thermal aggregates, i.e., a lower thermal stability.

[0052] [Test Example 1] To PG (final concentration 100 U / mL), component (A), 0.5 M of various amino acids was added to prepare an enzyme solution with a pH of 7.0 at 25°C. Separately, an enzyme solution (control; Ctrl) was prepared in the same manner, except that no amino acids were added. Each enzyme solution was heat-treated at 70°C for 15 minutes, then rapidly cooled to room temperature, and the absorbance at 413 nm was measured. The results for each enzyme solution are shown in Figure 1.

[0053] As shown in FIG. 1, when arginine (Arg), component (B), was added as an amino acid, the absorbance value significantly decreased, which indicated that the thermal stability of PG was significantly improved.

[0054] [Test Example 2] 0.1M, 0.5M, or 1.0M of arginine (Arg), component (B), was added to PG (final concentration 100 U / mL), component (A), to prepare a solution with a pH of 7.0 at 25°C. Separately, an enzyme solution (0M Arg) was prepared in the same manner, except that no arginine was added. Each prepared solution was heat-treated at 60°C for 30 minutes, and the absorbance at 413 nm was monitored during the heat treatment. The results are shown in Figure 2.

[0055] As shown in Figure 2, a decrease in absorbance was confirmed in the arginine-added sections (0.1 M Arg, 0.5 M Arg, and 1.0 M Arg) depending on the added arginine concentration, compared to the arginine-free section (0 M Arg). In other words, this result demonstrated that arginine, component (B), improves the thermal stability of PG.

[0056] [Test Example 3] 0.1 M of component (B) (arginine, argininamide, or spermidine) was added to component (A) PG (final concentration 100 U / mL) to prepare a solution with a pH of 7.0 at 25°C. Separately, an enzyme solution (control; Ctrl) was prepared in the same manner, except that component (B) was not added. Each prepared solution was heat-treated at 60°C for 10 minutes, and the absorbance at 413 nm was measured. The results are shown in Figure 3.

[0057] As shown in Figure 3, the results show that the stability of PG was significantly improved when arginine, argininamide, or spermidine was used. Specifically, when the absorbance value of the control was taken as 100%, the relative absorbance of the 0.1M arginine-added section was 70%, the relative absorbance of the 0.1M argininamide-added section was 62%, and the relative absorbance of the 0.1M spermidine-added section was 45%. In other words, the effect of spermidine in improving the thermal stability of PG was significantly more pronounced.

Claims

1. An enzyme preparation comprising (A) a protein deamidase and (B) a guanidium compound and / or a polyamine.

2. The enzyme preparation according to claim 1 , wherein the guanidinium compound is arginine and / or argininamide.

3. 3. The enzyme preparation according to claim 1 or 2, wherein the polyamine is spermidine.

4. The enzyme preparation according to any one of claims 1 to 3, wherein the content of the (B) component per 1 U of the (A) component is 0.5 µmol or more.

5. The enzyme preparation according to any one of claims 1 to 4, wherein the content of the component (B) is 0.05 M or more.

6. The enzyme preparation according to any one of claims 1 to 5, wherein the protein deamidase is protein glutaminase.

7. The enzyme preparation according to any one of claims 1 to 6, which is a liquid preparation.

8. 1. A method for deamidating a protein, comprising the step of treating a protein (P) with a protein deamidase (A) in the presence of a guanidium compound and / or a polyamine (B) to proceed with a deamidation reaction.

9. A method for producing a deamidated protein, comprising a step of proceeding a deamidation reaction of component (P) in a reaction mixture comprising a protein (P), (A) a protein deamidase, and (B) a guanidium compound and / or a polyamine.

10. The method according to claim 9 , wherein the reaction mixture is prepared by mixing the component (P) with an enzyme preparation containing the components (A) and (B).

11. The method according to claim 9 , wherein the reaction mixture is prepared by mixing a mixture containing the component (P) and the component (B) with the component (A).

12. The method according to any one of claims 9 to 11, wherein the deamidation reaction is carried out at a temperature higher than the optimum temperature of the component (A).

13. The method according to any one of claims 9 to 12, wherein the component (P) is a vegetable protein and / or an animal protein.

14. A method for producing a food, beverage, or pharmaceutical, comprising a step of proceeding a deamidation reaction of component (P) in a reaction mixture containing a food, beverage, or pharmaceutical raw material containing a protein (P), (A) a protein deamidase, and (B) a guanidium compound and / or a polyamine.

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