CN110623249B

CN110623249B - Instant enzyme preparation and preparation method thereof

Info

Publication number: CN110623249B
Application number: CN201911072996.6A
Authority: CN
Inventors: 王罗娜
Original assignee: Bu Guojian; Taixing Dongsheng Bio Tech Co ltd
Current assignee: Bu Guojian; Taixing Dongsheng Bio Tech Co ltd
Priority date: 2019-11-05
Filing date: 2019-11-05
Publication date: 2023-04-07
Anticipated expiration: 2039-11-05
Also published as: CN110623249A

Abstract

The invention relates to an instant enzyme preparation and a preparation method thereof. Specifically, the invention provides a quick-dissolving enzyme preparation which comprises glutamine transaminase, povidone and an effervescent disintegrant. The instant enzyme preparation can enable the TG enzyme to be dissolved completely and quickly and ensure the TG enzyme to have enough activity.

Description

Instant enzyme preparation and preparation method thereof

Technical Field

The invention relates to the field of bioactive enzyme preparations, and particularly provides an instant enzyme preparation and a preparation method thereof.

Background

The enzyme preparation is widely applied to the aspects of food (meat products, flour deep processing, bread baking industry, fruit processing industry and the like), textile, feed, lotion, paper making, leather, medicine, energy development, environmental protection and the like. Enzymes of biological origin are generally safer and can be used more safely as food additives, especially in food processing. The enzyme preparation industry is continuously developing, enriching and expanding to different application fields in China as a high-technology industry.

Transglutaminase (Transglutaminase), also known as Transglutaminase, is abbreviated as TGase or TG enzyme (hereinafter abbreviated as TG enzyme), widely exists in human bodies, higher animals, plants and microorganisms, is a natural source, and is an enzyme preparation for protein food developed by modern biotechnology. The TG enzyme is monomer protein with 331 amino groups and active center, has molecular weight of about 38000, and can catalyze acyl transfer reaction, so that covalent cross-linking in molecule or between molecules of protein polypeptide occurs, and the structure and function of the protein are improved. TG enzyme can improve foamability, adhesiveness, emulsibility, gel property, thickening property, emulsion stability and the like of protein, further improve appearance, flavor, taste, texture and the like of protein-rich food, and is widely applied to meat products, dairy products, fish products, bean products and flour products at present.

In industrial production, a large amount of TG enzyme is obtained mainly from microbial fermentation, and dry enzyme powder is finally obtained through a series of extraction, separation and purification. The prior TG enzyme preparation formulations are solid and liquid. However, in the production application of the existing solid TG enzyme preparation (such as powder or tablet), the TG enzyme preparation often cannot be completely dissolved after being added with water or is distributed unevenly in water, so that the utilization rate of the TG enzyme preparation is reduced, the application effect is poor, and the dosage of the TG enzyme preparation has to be increased. For the existing TG enzyme liquid preparation, the enzyme content is often greatly limited, most of the area and weight of the TG enzyme liquid preparation are water and glycerol substances, the transportation volume is large, the cost is high, and the enzyme activity preservation rate at normal temperature is not as long as that of a solid enzyme preparation.

Therefore, there is a need in the art to develop a preparation capable of rapidly and completely dissolving TG enzymes and ensuring that TG enzymes have sufficient activity.

Disclosure of Invention

The object of the present invention is to provide a preparation capable of dissolving TG enzyme rapidly and completely and ensuring sufficient activity of TG enzyme.

In a first aspect of the invention, there is provided a fast dissolving enzyme formulation comprising transglutaminase, povidone, and an effervescent disintegrant.

In another preferred embodiment, the weight ratio of glutamine transaminase to povidone is 1-10, preferably 2-8:1-6, more preferably 3-7:1-5, most preferably 4-6:2-4.

In another preferred embodiment, the weight ratio of glutamine transaminase to povidone is 0.5-5:1, preferably 0.8-3:1, more preferably 1-2.5, more preferably 1.2-2.0.

In another preferred embodiment, the povidone is present in an amount of 0.8-8wt%, preferably 1-6wt%, more preferably 1-5wt%, more preferably 1.5-4wt%, more preferably 2-3.5wt%, most preferably 2.4-2.8wt%, based on the total weight of the instant enzyme preparation.

In another preferred embodiment, the transglutaminase is present in an amount of 1-12wt%, preferably 1-10wt%, more preferably 1-8wt%, more preferably 2-6wt%, most preferably 3-5wt%, based on the total weight of the instant enzyme preparation.

In another preferred embodiment, the transglutaminase is present in an amount of 0.05 to 0.5 parts by weight, preferably 0.08 to 0.4 parts by weight, preferably 0.1 to 0.3 parts by weight, preferably 0.15 to 0.25 parts by weight, preferably 0.18 to 0.22 parts by weight.

In another preferred embodiment, the povidone is 0.02 to 1.2 parts by weight, preferably 0.05 to 1.0 parts by weight, more preferably 0.05 to 0.8 parts by weight, still more preferably 0.06 to 0.6 parts by weight, still more preferably 0.08 to 0.4 parts by weight, still more preferably 0.08 to 0.3 parts by weight, still more preferably 0.08 to 0.2 parts by weight, still more preferably 0.08 to 0.16 parts by weight, and most preferably 0.1 to 0.15 parts by weight.

In another preferred embodiment, said povidone is selected from the group consisting of: PVP90, PVP K30, or a combination thereof.

In another preferred embodiment, the effervescent disintegrant comprises an acid agent and an alkali agent.

In another preferred embodiment, the acid agent is selected from the group consisting of: an inorganic acid, an organic acid, or a combination thereof.

In another preferred embodiment, the organic acid has 1 to 10 carbon atoms in the molecular structure.

In another preferred embodiment, the acid agent is a C1-C10 lower organic acid.

In another preferred embodiment, the acid agent is selected from the group consisting of: citric acid, tartaric acid, malic acid, oxalic acid, boric acid, lactic acid, monopotassium phosphate, monosodium phosphate, tartaric acid, fumaric acid, or combinations thereof.

In another preferred example, the citric acid is citric acid with crystal water.

In another preferred embodiment, the citric acid is citric acid monohydrate.

In another preferred embodiment, the alkaline agent is selected from the group consisting of: carbonate, bicarbonate, or a combination thereof.

In another preferred embodiment, the carbonate is selected from the group consisting of: sodium carbonate, potassium carbonate, or a combination thereof.

In another preferred embodiment, the bicarbonate is selected from the group consisting of: sodium bicarbonate, potassium carbonate, or a combination thereof.

In another preferred embodiment, the alkaline agent is selected from the group consisting of: sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, or a combination thereof.

In another preferred embodiment, the acid agent is 0.1 to 2.5 parts by weight, preferably 0.2 to 2 parts by weight, preferably 0.5 to 2 parts by weight, more preferably 0.8 to 1.8 parts by weight, more preferably 1.0 to 1.6 parts by weight, more preferably 1.1 to 1.5 parts by weight, most preferably 1.2 to 1.4 parts by weight.

In another preferred embodiment, the alkaline agent is 0.4 to 4.0 parts by weight, preferably 0.5 to 3.0 parts by weight, more preferably 0.5 to 2.5 parts by weight, still more preferably 0.8 to 2.0 parts by weight, preferably 1.0 to 2.0 parts by weight, still more preferably 1.2 to 1.8 parts by weight, most preferably 1.4 to 1.6 parts by weight.

In another preferred embodiment, the amount of said acid agent is 10-50wt%, preferably 15-40wt%, more preferably 20-40wt%, most preferably 25-35wt%, based on the total weight of the instant enzyme preparation.

In another preferred embodiment, the alkaline agent is present in an amount of 10-50wt%, preferably 15-40wt%, more preferably 20-40wt%, most preferably 25-40wt%, based on the total weight of the instant enzyme preparation.

In another preferred embodiment, the instant enzyme preparation further comprises polyethylene glycol (PEG).

In another preferred embodiment, the molecular weight of the polyethylene glycol is 200-10000.

In another preferred embodiment, the polyethylene glycol is selected from the group consisting of: PEG200, PEG400, PEG600, PEG800, PEG1000, PEG1500, PEG2000, PEG3000, PEG4000, PEG6000, PEG8000, or combinations thereof.

In another preferred embodiment, the polyethylene glycol is PEG6000.

In another preferred embodiment, the polyethylene glycol is 0.05 to 1.0 part by weight, preferably 0.05 to 0.8 part by weight, more preferably 0.06 to 0.6 part by weight, more preferably 0.08 to 0.4 part by weight, more preferably 0.08 to 0.3 part by weight, more preferably 0.08 to 0.2 part by weight, more preferably 0.08 to 0.16 part by weight, most preferably 0.1 to 0.15 part by weight.

In another preferred embodiment, the polyethylene glycol is present in an amount of 0.8-8wt%, preferably 1-6wt%, more preferably 1-5wt%, more preferably 1.5-4wt%, more preferably 2-3.5wt%, most preferably 2.4-2.8wt%, based on the total weight of the instant enzyme preparation.

In another preferred embodiment, the instant enzyme preparation further comprises an inorganic salt.

In another preferred embodiment, the inorganic salt is selected from the group consisting of: potassium, sodium, calcium, magnesium, iron, zinc salts, or combinations thereof.

In another preferred embodiment, the inorganic salt is selected from the group consisting of: chloride salts, bromide salts, iodide salts, sulfate salts, phosphate salts, nitrate salts, or combinations thereof.

In another preferred embodiment, the inorganic salt is selected from the group consisting of: sodium chloride, potassium chloride, calcium chloride, magnesium chloride, ferric chloride, zinc chloride, sodium bromide, potassium bromide, calcium bromide, magnesium bromide, ferric chloride, zinc bromide, sodium iodide, potassium iodide, calcium bromide, magnesium bromide, ferric iodide, zinc iodide, sodium sulfate, potassium sulfate, calcium sulfate, magnesium sulfate, ferric sulfate, zinc sulfate, sodium phosphate, potassium phosphate, calcium phosphate, magnesium phosphate, iron phosphate, zinc phosphate, sodium nitrate, potassium nitrate, calcium nitrate, magnesium nitrate, ferric nitrate, zinc nitrate, or combinations thereof.

In another preferred embodiment, the inorganic salt is present in an amount of 5-40wt%, preferably 5-30wt%, more preferably 8-25wt%, more preferably 10-20wt%, most preferably 12-16wt%, based on the total weight of the instant enzyme preparation.

In another preferred embodiment, the inorganic salt is 0.1 to 4 parts by weight, preferably 0.2 to 3 parts by weight, more preferably 0.2 to 2 parts by weight, still more preferably 0.2 to 1.5 parts by weight, still more preferably 0.4 to 1.0 parts by weight, and most preferably 0.5 to 0.8 parts by weight.

In another preferred embodiment, the instant enzyme preparation further comprises a saccharide.

In another preferred embodiment, the saccharide is selected from the group consisting of: monosaccharides, disaccharides, or combinations thereof.

In another preferred example, the monosaccharide is a monosaccharide with a molecular structure containing 3-6 carbon atoms.

In another preferred example, the monosaccharide is a monosaccharide containing 5-6 carbon atoms in the molecular structure.

In another preferred embodiment, the monosaccharide is selected from the group consisting of: a five carbon sugar, a six carbon sugar, or a combination thereof.

In another preferred embodiment, the monosaccharide is selected from the group consisting of: arabinose, ribose, xylose, lyxose, glucose, mannose, fructose, galactose, or a combination thereof.

In another preferred embodiment, the disaccharide is selected from the group consisting of: maltose, lactose, sucrose, trehalose, or a combination thereof.

In another preferred embodiment, the saccharide is present in an amount of 0.1 to 4 parts by weight, preferably 0.2 to 3 parts by weight, more preferably 0.2 to 2 parts by weight, more preferably 0.2 to 1.5 parts by weight, more preferably 0.4 to 1.0 part by weight, most preferably 0.5 to 0.8 part by weight.

In another preferred embodiment, the carbohydrate is present in an amount of 5-40wt%, preferably 5-30wt%, more preferably 8-25wt%, more preferably 10-20wt%, most preferably 12-16wt%, based on the total weight of the instant enzyme preparation.

In another preferred embodiment, the instant enzyme preparation further comprises inorganic salts and saccharides.

In another preferred embodiment, the instant enzyme preparation is a solid preparation.

In another preferred embodiment, the solid preparation is a tablet.

In another preferred embodiment, the sum of the contents of all components of the instant enzyme preparation is 100wt%.

In another preferred embodiment, the instant enzyme preparation is dissolved in 200ml of water and the measured pH is 4.5-9.0, preferably 5.0-8.0, more preferably 5.0-7.0, more preferably 5.2-6.5, more preferably 5.5-6.2, more preferably 5.6-6.0, more preferably 5.7-5.9, more preferably 5.75-5.86, most preferably 5.78-5.87.

In another preferred embodiment, the instant enzyme preparation comprises 0.1-0.3 part by weight of glutamine transaminase, 0.08-0.2 part by weight of povidone K, 0.5-2 parts by weight of citric acid, 0.5-2.5 parts by weight of sodium bicarbonate, 0.2-1.5 parts by weight of glucose, 0.2-1.5 parts by weight of sodium chloride and 0.08-0.2 part by weight of PEG6000.

In another preferred example, the instant enzyme preparation comprises 0.15-0.25 parts by weight of glutamine transaminase, 0.08-0.16 parts by weight of povidone K, 1.0-1.6 parts by weight of citric acid, 1.0-2.0 parts by weight of sodium bicarbonate, 0.4-1.0 parts by weight of glucose, 0.4-1.0 parts by weight of sodium chloride and 0.08-0.2 parts by weight of PEG6000.

In another preferred embodiment, the instant enzyme preparation further comprises other pharmaceutically and food acceptable carriers.

In another preferred embodiment, the other pharmaceutically and food acceptable carrier is selected from the group consisting of: bacteriostatic agents, preservatives, colorants, antioxidants, opacifiers, or combinations thereof.

In a second aspect of the invention, there is provided a process for the preparation of a fast-dissolving enzyme preparation according to the first aspect of the invention, said process comprising the steps of: the components are mixed to obtain the instant enzyme preparation.

In another preferred embodiment, the components are mixed and then tabletted to obtain the instant enzyme preparation.

In another preferred embodiment, the components other than transglutaminase are mixed and then mixed with transglutaminase to obtain the instant enzyme preparation.

In a third aspect of the invention, there is provided the use of a fast-dissolving enzyme preparation according to the first aspect of the invention for improving the texture of a food product.

In another preferred embodiment, the food product is a protein-containing food product.

In another preferred embodiment, the food is a protein food.

In another preferred embodiment, the food is a meat food.

In another preferred embodiment, the food is a dairy product.

In another preferred embodiment, the food is a flour product.

In a fourth aspect of the present invention, there is provided a method for improving food quality, the method comprising: mixing a fast-dissolving enzyme preparation according to the first aspect of the invention with said food product to carry out an enzymatic cross-linking reaction.

In another preferred embodiment, the food is a protein food.

In another preferred example, the food is meat food.

In another preferred embodiment, the food is a dairy product.

In another preferred embodiment, the food is a flour product.

In another preferred embodiment, the method for improving the quality and structure of the food comprises the following steps: after mixing the instant enzyme preparation according to the first aspect of the invention with said food product, an enzymatic cross-linking reaction is carried out after addition of water.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.

Drawings

FIG. 1 is a graph showing the effect of crosslinking in example 5.

Detailed Description

The present inventors have made extensive and intensive studies and have unexpectedly developed a fast dissolving enzyme preparation comprising glutamine transaminase, povidone, and an effervescent disintegrant. In the instant enzyme preparation, the effervescent disintegrant can rapidly disperse the enzyme preparation, so that the TG enzyme is rapidly and completely dissolved, and the polyvidone can ensure that the TG enzyme still has higher enzyme activity in the enzyme preparation and after the enzyme preparation is rapidly dispersed, thereby avoiding the problem of significant reduction of the activity after the TG enzyme is prepared into a solid preparation. On the basis of this, the present invention has been completed.

Term(s)

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, the terms "comprises," "comprising," "includes," "including," and "including" are used interchangeably and include not only closed-form definitions, but also semi-closed and open-form definitions. In other words, the term includes "consisting of … …" and "consisting essentially of … …".

As used herein, the terms "PEG" and "polyethylene glycol" are used interchangeably.

As used herein, the terms "TG enzyme" and "transglutaminase" are used interchangeably.

Instant enzyme preparation

In order to overcome the defects that in the prior art, TG enzyme solid preparations are often incompletely dissolved by adding water or are unevenly dispersed in water, and the activity of TG enzyme is obviously reduced after TG enzyme is prepared into solid preparations and enzyme preparations are redissolved, the invention provides a quick-dissolving enzyme preparation, wherein the quick-dissolving enzyme preparation comprises (but is not limited to) glutamine transaminase, povidone and effervescent disintegrant. Preferably, the instant enzyme formulation further comprises one or more components selected from the group consisting of: polyethylene glycol, inorganic salt and saccharide.

In the instant enzyme preparation, the effervescent disintegrant can rapidly disperse the enzyme preparation, so that the TG enzyme is rapidly and completely dissolved, and the polyvidone can ensure that the TG enzyme still has higher enzyme activity in the enzyme preparation and after the enzyme preparation is rapidly dispersed, thereby avoiding the problem of remarkably reducing the activity after the TG enzyme is prepared into a solid preparation.

In a preferred embodiment of the invention, the weight ratio of glutamine transaminase to povidone is 1-10, preferably 2-8:1-6, more preferably 3-7:1-5, most preferably 4-6:2-4.

Preferably, the weight ratio of glutamine transaminase to povidone is 0.5-5:1, preferably 0.8-3:1, more preferably 1-2.5, more preferably 1.2-2.0.

In another preferred embodiment, the instant enzyme preparation is dissolved in 200ml of water and has a measured pH of 4.5-9.0, preferably 5.0-8.0, more preferably 5.0-7.0, more preferably 5.2-6.5, more preferably 5.5-6.2, more preferably 5.6-6.0, more preferably 5.7-5.9, more preferably 5.75-5.86, most preferably 5.78-5.87.

Glutamine transaminase

Transglutaminase (also called Transglutaminase), referred to as TGase or TG enzyme.

Preferably, in the instant enzyme preparation of the present invention, the transglutaminase is present in an amount of 0.05-0.5 parts by weight, preferably 0.08-0.4 parts by weight, preferably 0.1-0.3 parts by weight, preferably 0.15-0.25 parts by weight, preferably 0.18-0.22 parts by weight.

Preferably, the transglutaminase is present in an amount of 1-12wt%, preferably 1-10wt%, more preferably 1-8wt%, more preferably 2-6wt%, most preferably 3-5wt%, based on the total weight of the instant enzyme preparation.

Povidone

Povidone is also called polyvinylpyrrolidone (PVP) for short.

Representatively, the povidone includes (but is not limited to): PVP K90, PVP K30, or a combination thereof.

Preferably, in the instant enzyme preparation of the present invention, the povidone is 0.02 to 1.2 parts by weight, preferably 0.05 to 1.0 parts by weight, preferably 0.05 to 0.8 parts by weight, more preferably 0.06 to 0.6 parts by weight, more preferably 0.08 to 0.4 parts by weight, more preferably 0.08 to 0.3 parts by weight, more preferably 0.08 to 0.2 parts by weight, more preferably 0.08 to 0.16 parts by weight, most preferably 0.1 to 0.15 parts by weight.

Preferably, the content of povidone is 0.8-8wt%, preferably 1-6wt%, more preferably 1-5wt%, more preferably 1.5-4wt%, more preferably 2-3.5wt%, most preferably 2.4-2.8wt% based on the total weight of the instant enzyme preparation.

Effervescent disintegrant

In the invention, the effervescent disintegrant is a disintegrant which can generate gas effervescence by reacting with water, namely, the produced gas can rapidly disintegrate and disperse the preparation enzyme preparation, thereby rapidly and completely dissolving the TG enzyme.

In a preferred embodiment of the invention, the effervescent disintegrant comprises an acid agent and an alkali agent.

In another preferred embodiment, the effervescent disintegrant comprises an acid agent and reacts with the acid agent to produce CO ₂ Basic salts of gases (e.g., carbonates, bicarbonates, or combinations thereof).

In another preferred embodiment, the acid agent includes (but is not limited to): an inorganic acid, an organic acid, or a combination thereof;

preferably, the organic acid has 1 to 10 carbon atoms in the molecular structure.

Preferably, the acid agent is a C1-C10 lower organic acid.

Preferably, the acidic agent includes (but is not limited to): citric acid, tartaric acid, malic acid, oxalic acid, boric acid, lactic acid, monopotassium phosphate, monosodium phosphate, tartaric acid, fumaric acid, or combinations thereof.

Typically, the citric acid is citric acid with crystal water.

Typically, the citric acid is citric acid monohydrate.

In another preferred embodiment, the alkaline agent includes (but is not limited to): a carbonate, a bicarbonate, or a combination thereof.

Typically, the carbonate includes (but is not limited to): sodium carbonate, potassium carbonate, or a combination thereof.

Typically, the bicarbonate includes (but is not limited to): sodium bicarbonate, potassium carbonate, or a combination thereof.

Typically, the alkaline agents include (but are not limited to): sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, or a combination thereof.

Preferably, in the instant enzyme preparation of the present invention, the acid agent is 0.1 to 2.5 parts by weight, preferably 0.2 to 2 parts by weight, more preferably 0.5 to 2 parts by weight, still more preferably 0.8 to 1.8 parts by weight, still more preferably 1.0 to 1.6 parts by weight, still more preferably 1.1 to 1.5 parts by weight, most preferably 1.2 to 1.4 parts by weight.

Preferably, in the instant enzyme preparation of the present invention, the alkaline agent is 0.4 to 4.0 parts by weight, preferably 0.5 to 3.0 parts by weight, preferably 0.5 to 2.5 parts by weight, more preferably 0.8 to 2.0 parts by weight, preferably 1.0 to 2.0 parts by weight, more preferably 1.2 to 1.8 parts by weight, most preferably 1.4 to 1.6 parts by weight.

Optionally polyethylene glycol

In a preferred embodiment of the present invention, the instant enzyme preparation further comprises polyethylene glycol (PEG).

Preferably, the polyethylene glycol includes (but is not limited to): PEG200, PEG400, PEG600, PEG800, PEG1000, PEG1500, PEG2000, PEG3000, PEG4000, PEG6000, PEG8000, or combinations thereof.

Typically, the polyethylene glycol is PEG6000.

Preferably, in the instant enzyme preparation of the present invention, the polyethylene glycol is 0.05 to 1.0 part by weight, preferably 0.05 to 0.8 part by weight, more preferably 0.06 to 0.6 part by weight, more preferably 0.08 to 0.4 part by weight, more preferably 0.08 to 0.3 part by weight, more preferably 0.08 to 0.2 part by weight, more preferably 0.08 to 0.16 part by weight, most preferably 0.1 to 0.15 part by weight.

Optionally an inorganic salt

In a preferred embodiment of the present invention, the instant enzyme preparation further comprises an inorganic salt.

In another preferred embodiment, the inorganic salts include (but are not limited to): potassium, sodium, calcium, magnesium, iron, zinc salts, or combinations thereof.

In another preferred embodiment, the inorganic salts include (but are not limited to): chloride salts, bromide salts, iodide salts, sulfate salts, phosphate salts, nitrate salts, or combinations thereof.

Typically, the inorganic salts include (but are not limited to): sodium chloride, potassium chloride, calcium chloride, magnesium chloride, ferric chloride, zinc chloride, sodium bromide, potassium bromide, calcium bromide, magnesium bromide, ferric chloride, zinc bromide, sodium iodide, potassium iodide, calcium bromide, magnesium bromide, ferric iodide, zinc iodide, sodium sulfate, potassium sulfate, calcium sulfate, magnesium sulfate, ferric sulfate, zinc sulfate, sodium phosphate, potassium phosphate, calcium phosphate, magnesium phosphate, iron phosphate, zinc phosphate, sodium nitrate, potassium nitrate, calcium nitrate, magnesium nitrate, ferric nitrate, zinc nitrate, or combinations thereof.

Preferably, in the instant enzyme preparation according to the invention, the inorganic salt is present in an amount of 5-40wt%, preferably 5-30wt%, more preferably 8-25wt%, more preferably 10-20wt%, most preferably 12-16wt%, based on the total weight of the instant enzyme preparation.

In another preferred embodiment, the inorganic salt is 0.1 to 4 parts by weight, preferably 0.2 to 3 parts by weight, more preferably 0.2 to 2 parts by weight, still more preferably 0.2 to 1.5 parts by weight, still more preferably 0.4 to 1.0 part by weight, most preferably 0.5 to 0.8 part by weight.

Optionally a saccharide

In a preferred embodiment of the present invention, the instant enzyme preparation further comprises a saccharide.

In another preferred embodiment, the saccharides include (but are not limited to): monosaccharides, disaccharides, or combinations thereof.

Preferably, the monosaccharide is a monosaccharide with a molecular structure containing 3-6 carbon atoms.

Preferably, the monosaccharide is a monosaccharide with a molecular structure containing 5-6 carbon atoms.

Preferably, the monosaccharides include (but are not limited to): a five carbon sugar, a six carbon sugar, or a combination thereof.

Typically, the monosaccharides include (but are not limited to): arabinose, ribose, xylose, lyxose, glucose, mannose, fructose, galactose, or a combination thereof.

Typically, the disaccharides include (but are not limited to): maltose, lactose, sucrose, trehalose, or a combination thereof.

Preferably, in the instant enzyme preparation of the present invention, the saccharide is 0.1 to 4 parts by weight, preferably 0.2 to 3 parts by weight, more preferably 0.2 to 2 parts by weight, more preferably 0.2 to 1.5 parts by weight, more preferably 0.4 to 1.0 part by weight, most preferably 0.5 to 0.8 part by weight.

In another preferred embodiment, the saccharide is present in an amount of 5-40wt%, preferably 5-30wt%, more preferably 8-25wt%, more preferably 10-20wt%, most preferably 12-16wt%, based on the total weight of the instant enzyme preparation.

Optionally other pharmaceutically, food or nutraceutical acceptable carriers.

In a preferred embodiment of the present invention, the instant enzyme preparation further comprises other pharmaceutically, food or nutraceutical acceptable carriers.

As used herein, the term "pharmaceutically, food or nutraceutical acceptable carrier" refers to a substance that is suitable for use in humans and/or animals without undue adverse side effects (such as toxicity, irritation and allergic response), i.e., at a reasonable benefit/risk ratio.

In a preferred embodiment, the other pharmaceutically and food acceptable carriers include (but are not limited to): bacteriostatic agents, preservatives, colorants, antioxidants, opacifiers, or combinations thereof.

A preferred embodiment of the instant enzyme preparation of the present invention includes, but is not limited to, 0.1-0.3 parts by weight of transglutaminase, 0.08-0.2 parts by weight of povidone K, 0.5-2 parts by weight of citric acid, 0.5-2.5 parts by weight of sodium bicarbonate, 0.2-1.5 parts by weight of glucose, 0.2-1.5 parts by weight of sodium chloride, and 0.08-0.2 parts by weight of PEG6000.

Preferably, the instant enzyme preparation comprises (but is not limited to) 0.15-0.25 parts by weight of glutamine transaminase, 0.08-0.16 parts by weight of povidone K, 1.0-1.6 parts by weight of citric acid, 1.0-2.0 parts by weight of sodium bicarbonate, 0.4-1.0 parts by weight of glucose, 0.4-1.0 parts by weight of sodium chloride and 0.08-0.2 parts by weight of PEG6000.

Preparation method

The invention provides a preparation method of the instant enzyme preparation, which comprises the following steps: the components are mixed to obtain the instant enzyme preparation.

A preferred instant enzyme preparation is prepared as follows:

use of

The invention also provides the application of the instant enzyme preparation, which is used for improving the quality and structure of food.

In another preferred embodiment, the food product is a protein food product.

In another preferred example, the food is meat food.

In another preferred embodiment, the food is a dairy product.

In another preferred embodiment, the food is a flour product.

A method of enzymatic cross-linking reactions in food products.

The invention also provides a method for enzymatic cross-linking reactions in a food product, wherein the instant enzyme preparation according to the invention is mixed with the food product to carry out the enzymatic cross-linking reaction.

In another preferred embodiment, the method for enzymatic cross-linking reactions in a food product comprises the steps of: after mixing the instant enzyme preparation of the present invention with the food, water is added to carry out an enzymatic crosslinking reaction.

The main advantages of the invention include:

1. the invention unexpectedly develops an instant enzyme preparation which can not only rapidly and completely dissolve TG enzyme, but also ensure that the TG enzyme has enough efficacy, and avoid the problem that the activity of the TG enzyme is obviously reduced after the TG enzyme is prepared into a solid preparation.

2. The instant enzyme preparation can avoid the defects of powder scattering, difficult equipment cleaning, uneven distribution of TG enzyme and the like caused by a solid powder type TG enzyme preparation in the prior art.

3. The instant enzyme preparation has the advantages of small transportation space, low cost and lasting enzyme activity retention rate.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are by weight.

Example 1

Under an anhydrous and dry environment, 1.26g of citric acid monohydrate, 1.51g of sodium bicarbonate, 1.5g of TG enzyme complex (0.2 g of TG enzyme, 0.65g of anhydrous glucose, 0.65g of sodium chloride), 0.11g of PEG6000 and 0.12g of PVP K30 are weighed respectively. Respectively grinding citric acid and sheet PEG6000 into finer powder, and grinding and mixing the components except the TG enzyme compound uniformly; filling absolute ethyl alcohol into a 30ml sprinkling can, sprinkling twice in the mixed powder, quickly and uniformly mixing, and drying; mixing the TG enzyme complex in the previous step, and gently mixing uniformly; and (3) putting the obtained mixture into a groove of a tabletting machine for manual tabletting (19 mm), controlling the weight of each TG enzyme effervescent tablet with the diameter of 19mm to be 1.3g, drying the pressed TG enzyme effervescent tablet, and sealing and storing.

Example 2

Under an anhydrous and dry environment, 1.26g of citric acid monohydrate, 1.00g of sodium bicarbonate, 1.5g of TG enzyme complex (0.2 g of TG enzyme, 0.65g of anhydrous glucose, 0.65g of sodium chloride), 0.11g of PEG6000 and 0.12g of PVP K30 are weighed respectively. Respectively grinding citric acid and sheet PEG6000 into finer powder, and grinding and mixing the components except the TG enzyme compound uniformly; the absolute ethyl alcohol is filled in a 30ml sprinkling can, and is sprinkled twice in the mixed powder, and is rapidly and uniformly mixed and dried; mixing the TG enzyme complex in the previous step, and gently mixing uniformly; and (3) putting the obtained mixture into a groove of a tabletting machine for manual tabletting (19 mm and 5.5 mm), controlling the weight of each TG enzyme effervescent tablet with the diameter of 19mm to be 1.3g, controlling the weight of each TG enzyme effervescent tablet with the diameter of 5.5mm to be 0.1g, drying the pressed TG enzyme effervescent tablets, and sealing and storing.

Example 3

0.20g of citric acid monohydrate, 1.51g of sodium bicarbonate, 1.5g of TG enzyme complex (0.2 g of TG enzyme, 0.65g of anhydrous glucose, 0.65g of sodium chloride) and 0.11g of PEG6000 are weighed respectively in an anhydrous and dry environment. Respectively grinding citric acid and sheet PEG6000 into finer powder, and grinding and mixing the components except the TG enzyme compound uniformly; the absolute ethyl alcohol is filled in a 30ml sprinkling can, and is sprinkled twice in the mixed powder, and is rapidly and uniformly mixed and dried; mixing the TG enzyme complex in the previous step, and gently mixing uniformly; and (3) putting the obtained mixture into a groove of a tabletting machine for manual tabletting (19 mm), controlling the weight of each TG enzyme effervescent tablet with the diameter of 19mm to be 1.2g, drying the pressed TG enzyme effervescent tablet, and sealing and storing.

Comparative example 1 No sodium bicarbonate and PVP K30 was added

Under an anhydrous and dry environment, 1.26g of citric acid monohydrate, 1.5g of TG enzyme complex (0.2 g of TG enzyme, 0.65g of anhydrous glucose and 0.65g of sodium chloride) and 0.11g of PEG6000 are weighed respectively. Respectively grinding citric acid and sheet PEG6000 into finer powder, and grinding and mixing the components except the TG enzyme compound; the absolute ethyl alcohol is filled in a 30ml sprinkling can, and is sprinkled twice in the mixed powder, and is rapidly and uniformly mixed and dried; mixing the TG enzyme complex in the previous step, and gently mixing uniformly; and (3) putting the obtained mixture into a groove of a tabletting machine for manual tabletting (19 mm and 5.5 mm), controlling the weight of each TG enzyme effervescent tablet with the diameter of 19mm to be 1.0g, controlling the weight of each TG enzyme effervescent tablet with the diameter of 5.5mm to be 1g, drying the pressed TG enzyme effervescent tablets, and sealing and storing.

Comparative example 2 No PVP K30 addition

Under an anhydrous and dry environment, 1.26g of citric acid monohydrate, 1.51g of sodium bicarbonate, 1.5g of TG enzyme complex (0.2 g of TG enzyme, 0.65g of anhydrous glucose, 0.65g of sodium chloride) and 0.11g of PEG6000 are weighed respectively. Respectively grinding citric acid and sheet PEG6000 into finer powder, and grinding and mixing the components except the TG enzyme compound uniformly; filling absolute ethyl alcohol into a 30ml sprinkling can, sprinkling twice in the mixed powder, quickly and uniformly mixing, and drying; mixing the TG enzyme complex in the previous step, and gently mixing uniformly; and (3) putting the obtained mixture into a groove of a tabletting machine for manual tabletting (19 mm and 5.5 mm), controlling the weight of each TG enzyme effervescent tablet with the diameter of 19mm to be 1.3g, controlling the weight of each TG enzyme effervescent tablet with the diameter of 5.5mm to be 0.1g, drying the pressed TG enzyme effervescent tablets, and sealing and storing.

Comparative example 3

Under an anhydrous and dry environment, 1.16g of citric acid monohydrate, 1.61g of sodium bicarbonate, 1.5g of TG enzyme complex (0.2 g of TG enzyme, 1.30g of sodium chloride) and 0.11g of PEG6000 are weighed respectively. Respectively grinding citric acid and sheet PEG6000 into finer powder, and grinding and mixing the components except the TG enzyme compound uniformly; the absolute ethyl alcohol is filled in a 30ml sprinkling can, and is sprinkled twice in the mixed powder, and is rapidly and uniformly mixed and dried; mixing the TG enzyme complex in the previous step, and gently mixing uniformly; and (3) putting the obtained mixture into a groove of a tabletting machine for manual tabletting (19 mm), controlling the weight of each TG enzyme effervescent tablet with the diameter of 19mm to be 1.3g, drying the pressed TG enzyme effervescent tablet, and sealing and storing.

Comparative example 4

Under an anhydrous and dry environment, 0.80g of tartaric acid, 1.61g of sodium bicarbonate, 1.5g of TG enzyme complex (0.2 g of TG enzyme, 1.30g of sodium chloride) and 0.11g of PEG6000 are weighed respectively. Respectively grinding citric acid and sheet PEG6000 into finer powder, and grinding and mixing the components except the TG enzyme compound; filling absolute ethyl alcohol into a 30ml sprinkling can, sprinkling twice in the mixed powder, quickly and uniformly mixing, and drying; mixing the TG enzyme complex in the previous step, and gently mixing uniformly; and (3) putting the obtained mixture into a groove of a tabletting machine for manual tabletting (19 mm), controlling the weight of each 19mm TG enzyme effervescent tablet to be 1.3g, drying the pressed TG enzyme effervescent tablet, and sealing and storing.

Comparative example 5

0.63g of citric acid monohydrate, 0.63g of tartaric acid, 1.51g of sodium bicarbonate, 1.5g of TG enzyme complex (0.2 g of TG zymogen enzyme, 1.30g of sodium chloride) and 0.11g of PEG6000 are weighed in an anhydrous and dry environment respectively. Respectively grinding citric acid and sheet PEG6000 into finer powder, and grinding and mixing the components except the TG enzyme compound uniformly; the absolute ethyl alcohol is filled in a 30ml sprinkling can, and is sprinkled twice in the mixed powder, and is rapidly and uniformly mixed and dried; mixing the TG enzyme complex in the previous step, and gently mixing uniformly; and (3) putting the obtained mixture into a groove of a tabletting machine for manual tabletting (19 mm), controlling the weight of each TG enzyme effervescent tablet with the diameter of 19mm to be 1.3g, drying the pressed TG enzyme effervescent tablet, and sealing and storing.

Comparative example 6

Under an anhydrous and dry environment, 1.26g of citric acid monohydrate, 0.50g of sodium bicarbonate, 1.5g of TG enzyme complex (0.2 g of TG enzyme, 0.65g of anhydrous glucose, 0.65g of sodium chloride) and 0.11g of PEG6000 are weighed respectively. Respectively grinding citric acid and sheet PEG6000 into finer powder, and grinding and mixing the components except the TG enzyme compound uniformly; the absolute ethyl alcohol is filled in a 30ml sprinkling can, and is sprinkled twice in the mixed powder, and is rapidly and uniformly mixed and dried; mixing the TG enzyme complex in the previous step, and gently mixing uniformly; and (3) putting the obtained mixture into a groove of a tabletting machine for manual tabletting (19 mm), controlling the weight of each TG enzyme effervescent tablet with the diameter of 19mm to be 1.2g, drying the pressed TG enzyme effervescent tablet, and sealing and storing.

Example 4 effect test:

this example considers the effervescent effect and enzymatic activity of TG enzyme effervescent tablets prepared in examples 1-3 and comparative examples 1-6

Effervescent effect

The above TG enzyme effervescent tablets of examples 1-3 and comparative examples 1-6 were tested for effervescence and pH in 200ml water, and the statistical results are shown in Table 1:

table 1 examples 1-3 comparative examples 1-6 effervescence and pH testing

As can be seen from Table 1, the TG enzyme effervescent tablets prepared in examples 1 to 3 had excellent effervescent effect and were rapidly and completely dispersed and dissolved.

Enzyme Activity assay

The above-mentioned TG enzyme effervescent tablets of examples 1 to 3 and comparative examples 1 to 6 were examined for enzyme activity.

The enzymatic activity of the TG enzyme is calculated by measuring the amount of hydroxamic acid generated by the reaction of CBZ-Gln-Gly and hydroxylamine by a colorimetric method, and the specific operation steps are as follows:

preparation of reagents:

(1) Substrate solution a: 2.42g Tris,0.7g hydroxylamine hydrochloride, 0.31g reduced glutathione, 1.01g CBZ-Gln-Gly are weighed out accurately and dissolved in 80ml water, pH is adjusted to 6.0 by 6N hydrochloric acid, finally the volume is fixed to 100ml by water, and the solution needs to be stored in a refrigerator at 4 ℃ and is effective within one week.

(2) Color developing solution B:

solution 1:3N hydrochloric acid

Solution 2:12g of the composition was dissolved in 100ml of water

Solution 3:5g of ferric chloride hexahydrate are dissolved in 100ml of 0.1N hydrochloric acid

Before use, the solution 1,2,3 is mixed in equal proportion to obtain the developing solution.

(3) Standard curve:

a. 64.8mg of L-glutamic acid-gamma-monohydroxamic acid was weighed and dissolved in 10ml of 0.2mol/L Tris-HCl buffer (pH 6.0) to obtain a standard solution.

b. Diluting the standard solution with 0.2mol/L Tris-HCl buffer solution to 8.0, 16.0, 20.0, 24.0 and 32.0 mu mol L-glutamic acid-gamma-monohydroxamic acid per ml;

c. respectively putting 0.2ml of the standard solution into test tubes, accurately adding 2ml of substrate solution into each test tube, oscillating and mixing uniformly, carrying out water bath at 37 +/-1 ℃ for 10min, and adding 2ml of color developing solution.

d. Removing precipitate with the same detection method, and measuring the absorbance of the supernatant with water at 525nm as control;

e. a standard curve relating the absorption Ab to the concentration Co (. Mu.mol/ml) of L-glutamic acid-gamma-monohydroxamic acid can be drawn according to the method described above, ab =8.831Co-0.0456, correlation coefficient R ² ＝0.9999。

(4) And (3) sample determination:

A. calculating the content of TG enzyme in each prepared effervescent tablet, and preparing into a solution containing TG enzyme of 1 mg/ml.

B. Diluting the solution in the step A by 10 times: 100. Mu.l of the stock solution was dissolved in 900. Mu.l of water.

C. Preparing test tubes, respectively sucking 2ml of the reagent A by a pipette gun, adding into a reaction tube, adding 2ml of the reagent B into the blank, and carrying out water bath at 37 ℃ for 5min;

D. accurately sucking 200 μ l of sample to be detected, adding into the tube to be reacted respectively, adding 200 μ l of sample into blank tube, mixing for 5 s with oscillator, and water-bath at 37 deg.C for 10min;

E. adding 2ml of the reagent B into the reaction tube, adding 2ml of the reagent A into the blank tube, and uniformly mixing for 5 seconds by using an oscillator.

F. Each tube was removed from 1ml to 2ml of EP tube and centrifuged at 3000rpm for 10min.

G. Color comparison: and (4) transferring 1000 mu l of the solution to be detected into a cuvette by taking the blank as a control, reading in an ultraviolet spectrophotometer with the wavelength of 525nm, and recording the light absorption value.

5) And (3) calculating enzyme activity:

U/g＝(C·D)/(W·10)

c: the concentration of the corresponding hydroxamic acid (. Mu. Mol/ml) was determined from the standard curve

D: dilution factor (ml) of test solution

W: sample weight to be measured (g)

10: reaction time (min)

Definition of enzyme activity: one unit of enzyme activity is defined as the amount of enzyme that catalyzes the production of 1. Mu. Mol hydroxamic acid per minute of the substrate under the above reaction conditions.

Examples 1-3 the results of measuring the specific enzyme activity of the TG enzyme effervescent tablets of comparative examples 1-6 are shown in Table 2:

TABLE 2 specific enzyme activity (n =3,M. + -. SD)

As can be seen from Table 2, the effervescent tablets prepared in examples 1 to 3 had higher enzyme activity after being rapidly dispersed, as compared with comparative examples 1 to 6.

Example 5

This example examines the effect of the TG enzyme effervescent tablet prepared in example 1 on the crosslinking of the meat paste.

Experimental methods

(1) Taking out the meat paste at the temperature of minus 20 ℃ and then melting the meat paste, weighing 10g of each part, putting the weighed parts into a beaker, and weighing three parts in total, wherein the numbers of the parts are 1,2 respectively;

(2) Number 1 is blank control, no TG enzyme is added, and the meat is stirred into meat balls; number 2, adding a TG enzyme effervescent tablet prepared in example 1, adding 2ml of water as an effervescent tablet solvent, stirring to wrap the meat paste, and stirring to form meat balls after the effervescent tablet is finished;

(3) Numbering 1-2, reacting at room temperature for 1h;

(4) After the reaction is finished, cooking the meat balls with the numbers of 1-2 on an induction cooker;

(5) The crosslinking effect was observed and is shown in FIG. 1.

The experimental results are as follows:

as can be seen from fig. 1: after the enzyme reaction is finished, the serial number 2 is crosslinked and agglomerated before the enzyme reaction is cooked; the TG enzyme effervescent tablets of example 1 had excellent crosslinking effect on the meat paste, and the cooked meat paste was crosslinked into a mass having more elasticity and better taste.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims

1. A fast-dissolving enzyme preparation, characterized in that the fast-dissolving enzyme preparation comprises the following components by weight:

and the sum of the contents of all the components of the instant enzyme preparation is 100wt%;

wherein, the instant enzyme preparation is dissolved in 200ml of water, and the measured pH value is 5.0-6.0.

2. The instant enzyme preparation according to claim 1, wherein the PVP K30 is present in an amount of 2.4-2.8wt%, based on the total weight of the instant enzyme preparation.

3. The instant enzyme formulation of claim 1, wherein the PEG6000 is present in an amount of 2.4 to 2.8wt% based on the total weight of the instant enzyme formulation.

4. The instant enzyme formulation according to claim 1, wherein the sodium chloride is present in an amount of 12-16wt% based on the total weight of the instant enzyme formulation.

5. The instant enzyme preparation according to claim 1, wherein the glucose is present in an amount of 12-16 wt.%, based on the total weight of the instant enzyme preparation.

6. The instant enzyme formulation according to claim 1, wherein the instant enzyme formulation is dissolved in 200ml of water and has a measured pH of 5.7-5.9.

7. The instant enzyme formulation according to claim 1, wherein the instant enzyme formulation is dissolved in 200ml of water and has a measured pH of 5.75-5.86.

8. The instant enzyme formulation according to claim 1, wherein the instant enzyme formulation is dissolved in 200ml of water and has a measured pH of 5.78-5.87.

9. The instant enzyme preparation according to claim 1, wherein the instant enzyme preparation is 1.26g of citric acid monohydrate, 1.51g of sodium bicarbonate, 0.2g of glutamine transaminase, 0.65g of anhydrous glucose, 0.65g of sodium chloride, 0.110.11g of PEG6000, 0.12g of PVP K30.

10. The instant enzyme preparation according to claim 1, wherein the instant enzyme preparation is 1.26g of citric acid monohydrate, 1.00g of sodium bicarbonate, 0.2g of glutamine transaminage, 0.65g of anhydrous glucose, 0.65g of sodium chloride, 0.11g of PEG6000, 0.12g of PVP K30.

11. The method of preparing the instant enzyme formulation of claim 1, comprising the steps of: the components are mixed to obtain the instant enzyme preparation.

12. Use of the instant enzyme preparation according to claim 1 for improving the quality of food texture.

13. A method for improving the texture of a food product, characterized in that an instant enzyme preparation according to claim 1 is mixed with said food product and subjected to an enzymatic cross-linking reaction.