Liquid enzyme preparation and preparation method thereof

A technology of liquid enzymes and preparations, which is applied in the field of transglutaminase (EC.2.3.2.13) liquid preparations and preparations, liquid enzyme preparations and preparations, and can solve problems such as loss, air leakage, cumbersome nitrogen filling operations, etc.

Active Publication Date: 2016-04-06
SHANGHAI KINRY FOOD INGREDIENTS +1
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1) Production cost: In the production process of transglutaminase, the fermented liquid obtained first, and the process of refining the enzyme liquid require precipitation, drying, crushing and other processing steps, and the process is complicated, resulting in an increase in production cost;
[0006] 2) Safety protection: Transglutaminase powder preparations are easy to spread during production and use, resulting in poor workshop environment and difficult cleaning of equipment; the operation space is full of dust, making protection difficult. After the enzyme powder is inhaled by the human body, it will cause a strong allergic reaction , seriously damage the respiratory organs of the operator, and symptoms such as high fever, pneumonia, and shock appear, which pose a great hazard to the health of the operator. So far, transglutaminase has repeatedly occurred in the factory and caused allergic symptoms such as cough and fever. event;
[0007] 3) Packaging problem: The transglutaminase powder enzyme preparation is vacuum-packed, and the powder is easily sucked into the equipment during vacuuming, causing certain losses; if the enzyme preparation packaging is not tightly sealed, resulting in air leakage, it is prone to moisture absorption and agglomeration , reduced enzyme activity, etc. If nitrogen-filled packaging is used, the nitrogen-filled operation will be cumbersome and cause a lot of dust, and the production cost will increase significantly;
And it often happens that adding water does not completely dissolve or directly adding powder is prone to uneven distribution. At the same time, when using equipment for rolling and kneading operations, it is easy to stick to the equipment, resulting in reduced utilization of powder preparations and poor application effects, so it has to be added The dosage of transglutaminase powder preparation;
[0009] 5) The development trend of enzyme preparations: In biological enzyme preparations and various industrial fields, many types of enzyme preparations have adopted liquid dosage forms, and solved problems in the process of storage, transportation and application, and realized economical and green production and use. Such as liquid cellulase (patent publication number: CN103981168A, CN101381716), liquid phytase (patent publication number: CN101617740), liquid pectinase (patent publication number: CN104531653A), liquid lipase (patent publication number: CN103525797A), etc. , but there is no commercialized liquid preparation of transglutaminase for long-term stable storage at room temperature in the world
[0010] Patent Publication No. CN104024406A provides a method for manufacturing a liquid enzyme preparation of glutamyl transaminase. In view of the fact that the method has a maximum storage period of 180 days at low temperature and needs to be stored and transported under frozen conditions, the commercial use of this method is limited. , so far no finished products using this method have been seen

Method used

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  • Liquid enzyme preparation and preparation method thereof
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  • Liquid enzyme preparation and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] The preparation of transglutaminase crude liquid uses microbial fermentation method, which can refer to the patent (patent publication number: EP0379606B1): using Streptomycesmobaraensis as the starting strain, use an inoculation loop to pick a few well-growing colonies and inoculate them on the slope culture medium at 30°C for 7 days. The activated strains are then inserted into the seed medium, cultivated at 30°C for 48 hours, added to the fermentation medium according to 10% inoculum, and cultivated at 30°C for 48-72 hours to obtain a crude transglutaminase solution, enzyme The purification and concentration of the crude liquid is carried out according to the following steps:

[0108] 1) Press filter: the pore size is 1 μm, the operating pressure is 0.20 MPa, the temperature is 20 ° C, and the filtrate is taken;

[0109] 2) Microfiltration: the pore size is 0.25μm, the operating pressure is 0.25MPa, the temperature is 20°C, and the filtrate is taken;

[0110] 3) Ul...

Embodiment 2

[0115] The crude liquid of transglutaminase is prepared by enzyme powder dissolving method. Weigh a certain amount of pure water, put it in a capacity with stirring function, and make the enzyme activity 5000-8000u / ml of transglutaminase powder (Shanghai Qingrui Food Co., Ltd. Enzyme powder produced by Science and Technology Co., Ltd.) was slowly added, stirred while adding to dissolve the enzyme solution with a certain concentration, and then sterilized by microfiltration to obtain a clear transglutaminase solution with an enzyme activity of 2500u / ml and an enzyme activity of 2500u / ml transglutaminase purified and concentrated enzyme solution. The detected pH is 6.30, the water activity is 0.95, and the oxidation-reduction potential is 58mv, and it is ready for use.

Embodiment 3

[0117] A preparation method of liquid transglutaminase, the total weight of the liquid enzyme is 1000ml, the preparation method is as follows, weighed according to the ingredients list 11,

[0118] Table 11. Preparation of a liquid transglutaminase

[0119]

[0120] Weigh the materials in the formula, mix evenly, and filter through a 0.1 micron sterile membrane to obtain a light yellow solution. The redox potential is -100mv, the water activity is 0.79, the pH is 6.0, and the enzyme activity is 498.1u / ml. Fill in 10 bottles of 100mL PET opaque bottles, and finally prepare the liquid transglutaminase preparation. After storing at room temperature for 180 days, observe the enzyme preparation of this formula, detect the enzyme activity, and calculate the preservation rate of the enzyme activity. GB4789.2-2010 National Food Safety Standard "Determination of Total Bacterial Colony in Food Microbiology Inspection", GB4789.3 National Food Safety Standard "Coliform Count in Food Mi...

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Abstract

The invention discloses a liquid enzyme preparation and a preparation method thereof and mainly solves the technical problem that a glutamine transaminase liquid preparation is hard to preserve at the normal temperature. According to the technical scheme, the enzyme preparation is a liquid preparation of glutamine transaminase EC2.3.2.13, and the constituent content of the enzyme preparation is that enzyme activity of the glutamine transaminase liquid preparation is 10-1000 u/ml, the weight volume percentage content of a water activity conditioning agent is 30-80%, the weight volume percentage content of a redox potential conditioning agent is 0.0075-1%, the weight volume percentage content of a food preservative is 0-0.1%, and a pH conditioning agent is supplemented to 100% in volume. The preparation method of the liquid enzyme preparation comprises the steps of enzyme liquid purification, mixing, sterilization and bottling to obtain a finished product. By means of the liquid enzyme preparation and the preparation method thereof, the enzyme activity of the enzyme preparation preserved for six months at the normal temperature can be maintained at 80% or above, the character is stable, no microbial spoilage is caused, and the using effect is good.

Description

technical field [0001] The invention discloses a liquid enzyme preparation and a preparation method, in particular to a liquid preparation and preparation method of transglutaminase (EC. The invention belongs to the field of enzyme preparation and the field of food additives. Background technique [0002] Glutamine transaminase (EC.2.3.2.13, English full name Amineγ-glutamyl-transferase, English abbreviation Transglutaminase, abbreviated name TG) is a protein cross-linking enzyme, which can use the ε-amino group of lysine in the protein peptide chain as Acyl acceptors, catalyze the formation of ε-(γ-glutamyl) lysine hetero-peptide bonds within and between molecules of proteins, improve the emulsification, rheology, solubility, foaming and other properties of proteins, endow The unique texture properties and adhesiveness of protein can improve the color and fragrance of the product, which has great market value. [0003] Transglutaminase was first isolated from guinea pig l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/96C12N9/10
CPCC12N9/1044C12N9/96C12Y203/02013
Inventor 王方明谢永强徐保弟张宇
Owner SHANGHAI KINRY FOOD INGREDIENTS
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