CN106281935A - A kind of method promoting neutral protease activity and the application in beer clarification thereof - Google Patents

Abstract

The invention relates to the field of food processing, in particular to a method for promoting the activity of neutral protease and its application in beer clarification. The method for promoting neutral protease activity and clarifying beer thereof according to the present invention is to use enzyme promoter 8-((4-(2,4-dimethoxyphenyl)piperazine-1-yl)methyl) ‑5,7‑Dihydroxy‑3‑(4‑hydroxyphenyl)‑4H‑chromen‑4‑one is added to puree beer with neutral protease to prepare clarified beer. The neutral protease accelerator of the present invention improves the activity of neutral protease, improves the clarification effect, and reduces the usage of neutral protease; the present invention can ensure the high clarity of beer in the process of transportation and refrigeration, and can also improve the taste of beer , improve the content of polypeptide in beer, and increase the nutritional value of beer.

Description

A method for promoting the activity of neutral protease and its application in beer clarification

technical field

The invention relates to the field of food processing, in particular to a method for promoting the activity of neutral protease and its application in beer clarification."

technical background

Neutral protease, also known as serrapeptase, has a molecular weight of 35-40KD. It is a protease with high activity under neutral conditions. It can be used for enzymatic hydrolysis of various proteins. As a biocatalyst, it has fast catalytic reaction speed, No industrial pollution and other advantages. According to the provisions of "GB 2760-2011 National Food Safety Standard - Standards for the Use of Food Additives", neutral protease can be used as a processing aid (enzyme preparation) in the food industry."

During the transportation and refrigeration of beer, due to the effects of light, vibration, temperature and other environmental conditions, turbidity and precipitation will occur. The cause of beer turbidity and precipitation is the combination of residual protein and polyphenols in beer to form macromolecular complexes. thing. Treating raw beer with neutral protease can decompose macromolecular protein in beer. This method is fast, simple and effective, and has important application value in production. However, the high price of neutral protease has affected the production cost of beer and the application field of neutral protease".

8–((4–(2,4–dimethoxyphenyl)piperazin–1–yl)methyl)–5,7–dihydroxy–3–(4–hydroxyphenyl)–4H–benzo Pyran-4-one is a genistein derivative obtained by chemical synthesis of genistein and 1-(2,4-dimethylphenyl)piperazine. 8–((4–(2,4–dimethoxyphenyl)piperazin–1–yl)methyl)–5,7–dihydroxy–3–(4–hydroxyphenyl)–4H–benzo The molecular formula of pyran-4-one is C ₂₈ H ₂₈ N ₂ O ₅ , the molecular weight is 472.52, and its structural formula is as follows."

At present, there is no 8-((4-(2,4-dimethoxyphenyl)piperazin-1-yl)methyl)-5,7-dihydroxy-3-(4-hydroxyphenyl )-4H-benzopyran-4-one application report in promoting the activity of neutral protease".

Contents of the invention

The object of the present invention is to provide a method using 8-((4-(2,4-dimethoxyphenyl)piperazin-1-yl)methyl)-5,7-dihydroxy-3-(4- Hydroxyphenyl)-4H-benzopyran-4-one is used as an enzyme promoter to promote the activity of neutral protease, which not only improves the clarification effect of beer, but also obtains clarified beer with higher nutritional value."

The method for promoting neutral protease activity of the present invention is to use enzyme promoter 8-((4-(2,4-dimethoxyphenyl)piperazin-1-yl)methyl)-5,7 -Dihydroxy-3-(4-hydroxyphenyl)-4H-benzopyran-4-one was added to raw beer with neutral protease to prepare clarified beer".

in:

The addition amount of the neutral protease accelerator is 2-4% of the mass of the neutral protease, and the addition amount of the neutral protease is 4-5 mg per liter of raw beer."

The enzymolysis temperature is 35-40°C, and the enzymolysis time is 32-34h".

After enzymatic hydrolysis, use a hollow fiber ultrafiltration membrane with a molecular weight cutoff of 20,000 for ultrafiltration."

Neutral protease promoter 8–((4–(2,4–dimethoxyphenyl)piperazin–1–yl)methyl)–5,7–dihydroxy–3–(4–hydroxyphenyl) -4H-benzopyran-4-one is a chemical compound of genistein and 1-(2,4-dimethylphenyl)piperazine, and the specific preparation process is as follows."

Add 1mmol of genistein, 25mL of absolute ethanol and 0.08mL of 37% formaldehyde solution into a 50mL round bottom flask, heat to 55-60°C, after the reaction solution is clarified, add 1-(2,4-dimethylphenyl) Piperazine 0.19mL, stirred at room temperature for 48h, a solid precipitated, filtered, and the solid was separated and purified by recrystallization to obtain the enzyme accelerator of the present invention".

The neutral protease promoter is combined with a tyrosine residue of the neutral protease by a hydrogen bond, two π bonds are combined with a lysine residue, and three π bonds are combined with a tryptophan residue. This combination results in a conformational change of the dispase, which increases the activity of the dispase".

Compared with the existing technology, the beneficial effects of the present invention are as follows".

(1) The neutral protease accelerator of the present invention improves the activity of neutral protease, and it is added to the raw beer together with neutral protease, which has a synergistic effect, reduces the amount of neutral protease used, and improves the clarification of beer The effect ensures the high clarity of beer during transportation and refrigeration."

(2) The production method of the present invention improves the taste of beer, increases the content of polypeptides, and increases the nutritional value of beer".

detailed description

The present invention will be further described below in conjunction with embodiment ".

Example 1

In the raw beer, add 5 mg/L of neutral protease, and at the same time add a neutral protease accelerator 8–((4–(2,4–dimethoxyphenyl)piperazine– 1-yl)methyl)-5,7-dihydroxy-3-(4-hydroxyphenyl)-4H-benzopyran-4-one, shaken in an air bath at 35°C for 34h, enzymolysis After the clarification is completed, the hollow fiber ultrafiltration membrane is ultrafiltered to obtain clarified beer".

After adding the neutral protease accelerator, the clarity increased by 8% compared with the beer without the neutral protease accelerator, and the clarity increased by 5% compared with the non-use neutral protease accelerator. Peptide content was 4.5% and 1.1% higher in beers with and without dispase booster than in beer without dispase”.

Example 2

In the raw beer, add 4mg/L of neutral protease, and at the same time add the neutral protease accelerator 8–((4–(2,4–dimethoxyphenyl)piperazine– 1-yl)methyl)-5,7-dihydroxy-3-(4-hydroxyphenyl)-4H-benzopyran-4-one, shaken in an air bath at 40°C for 32h, enzymolysis After the clarification is completed, the hollow fiber ultrafiltration membrane is ultrafiltered to obtain clarified beer".

Compared with the beer without neutral protease, the clarity increased by 11% after adding neutral protease accelerator, and the clarity increased by 8% compared with that without neutral protease accelerator. Peptide content was 4.5% and 1.1% higher in beers with and without dispase booster than in beer without dispase”.

Claims (5)

1. one kind promotes the method for neutral protease activity and the application in beer clarification thereof, it is characterised in that: neutral protein Enzyme accelerator 8 ((4 (2,4 Dimethoxyphenyl) piperazine 1 base) methyl) 5,7 dihydroxy 3 (4 hydroxy phenyl) 4H .alpha.-5:6-benzopyran 4 ketone adds in plasmogen beer together with neutral protease, the medicated beer of preparation clarification.

A kind of method promoting neutral protease activity the most according to claim 1 and the application in beer clarification thereof, It is characterized in that: the addition of enzyme accelerator is the 2～4% of neutral protease quality, the addition of neutral protease be every liter former Slurry medicated beer adds 4～5mg neutral protease.

A kind of method promoting neutral protease activity the most according to claim 1 and the application in beer clarification thereof, It is characterized in that: hydrolysis temperature is 35～40 DEG C, enzymolysis time is 32～34 h.

A kind of method promoting neutral protease activity the most according to claim 1 and the application in beer clarification thereof, It is characterized in that: in plasmogen beer, add neutral protease and neutral protease accelerator, after enzymolysis, ultrafiltration Medicated beer to clarification；Than not using the medicated beer ratio of neutral protease after adding neutral protease accelerator, clarity improve 8～ 11%, improve 5～8% than not using neutral protease accelerator to compare clarity；Use and do not use neutral protease to promote The medicated beer of agent improves content of peptides than the clarity of the medicated beer not using neutral protease and increases 4.5～8.3% He respectively 1.1～3.2%.

A kind of method promoting neutral protease activity the most according to claim 4 and the application in beer clarification thereof, It is characterized in that: use and block the hollow fiber ultrafiltration membrane ultrafiltration that molecular weight is 20000.