Method for preparing immobilized fructosyltransferase through ion exchange fibers

A technology of fructosyltransferase and ion exchange fiber, which is applied in the preparation of immobilized fructosyltransferase by using ion exchange fiber, and in the field of immobilized fructosyltransferase by ion exchange fiber, which can solve the problem of poor mechanical resistance of immobilized enzyme and immobilized enzyme Short half-life, food safety risks and other issues, to achieve the effect of low fiber loss, high cycle times, and mature equipment

Inactive Publication Date: 2015-03-25
LINYI CITY HONGYU BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The above immobilized fructosyltransferases have better industrial application prospects than free enzymes, and realize the characteristics of recyclable and repeated use. However, the above methods are complicated to prepare, the activity after immobilization is not high, and the half-life of the immobilized enzyme is short. , Using more chemical reagents, using toxic glutaraldehyde as a cross-linking agent, due to the enzyme falling off in the middle of the reaction process, the free glutaraldehyde will cause food safety risks to the product, and the mechanical resistance of immobilized enzymes is poor.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Aspergillus oryzae (Aspergillus oryzae) was fermented, separated and washed under the conditions of temperature = 28°C, pH = 4.5, time = 36h, ventilation rate 0.15VVM, rotation speed = 150r / min. times (W / W) weight of purified water and 1.5% (W / W) of immobilized cellulase with an activity of 100,000 U / g, stirring speed = 50r / min, operating temperature = 30°C, reacting for 5 hours, using centrifugal radius = 400mm, motor speed = 1400r / min centrifuge to separate the liquid to obtain free fructosyltransferase and put it in an immobilized reaction tank, add 1g of food-grade ethanol with concentration = 55% (V / V) per enzyme activity = 300U Weakly alkaline anion exchange fibers soaked for 40 minutes and washed twice with purified water. Immobilization temperature = 25°C, pH value = 5.5, stirring speed = 35r / min, adsorption time = 2h, and then filter the solid to obtain immobilized fructosyltransferase; the obtained immobilized fructosyltransferase is stored in a cold storage a...

Embodiment 2

[0020] Aspergillus oryzae was fermented, separated and washed under the conditions of temperature = 33°C, pH = 5.5, time = 42h, ventilation rate 0.15VVM, rotation speed = 150r / min. times (W / W) weight of purified water and 2.0% (W / W) of immobilized cellulase with an activity of 150,000 U / g, stirring speed = 50r / min, operating temperature = 30°C, reacting for 8 hours, using centrifugal radius = 400mm, motor speed = 1500r / min centrifuge to separate the liquid to obtain free fructosyltransferase and put it in the immobilized reaction tank, add 1.5g per enzyme activity = 400U and use concentration = 70% (V / V) food grade Soak in ethanol for 60min and wash 4 times with purified water for negative weak alkaline ion exchange fiber. Immobilization temperature = 28°C, PH value = 6.5, stirring speed = 45r / min, adsorption time = 3h, and then filter the solid to obtain immobilized fructosyltransferase; the obtained immobilized fructosyltransferase is stored in a 5°C freezer 6h, washed with...

Embodiment 3

[0022] Aspergillus oryzae (Aspergillus oryzae) was fermented, separated and washed under the conditions of temperature = 31°C, pH = 5.5, time = 38h, ventilation rate 0.15VVM, rotation speed = 150r / min. times (W / W) weight of purified water and 2.0% (W / W) of immobilized cellulase with an activity of 120,000 U / g, stirring speed = 60r / min, operating temperature = 30°C, reacting for 5 hours, using centrifugal radius = 400mm, motor speed = 1500r / min centrifuge to separate the liquid to obtain free fructosyltransferase and put it in an immobilized reaction tank, add 1.2g per enzyme activity = 320U and use concentration = 60% (V / V) food grade Soak in ethanol for 45min and wash 3 times with purified water for negative weak alkaline ion exchange fiber. Immobilization temperature = 26°C, PH value = 6.0, stirring speed = 40r / min, adsorption time = 3h, and then filter the solid to obtain immobilized fructosyltransferase; the obtained immobilized fructosyltransferase is stored in a 4°C freeze...

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PUM

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Abstract

The invention provides a method for preparing immobilized fructosyltransferase through ion exchange fibers. The weakly basic anion exchange fibers are used as a carrier to immobilize a liquid free fructosyltransferase to the carrier, the activity of the immobilized fructosyltransferase can be maintained for a long time, and the immobilized fructosyltransferase can be repeatedly used for a long time. The method comprises two pieces of technological content, namely the liquid free fructosyltransferase is prepared through biolysis and the immobilized fructosyltransferase is prepared through the ion exchange fibers. Fermentation cultivation is conducted on aspergillus oryzae to obtain a large amount of thallus, the thallus is washed, celluloses are used for dissolving the thallus, free enzyme fluid is extracted, and then the fructosyltransferase is immobilized through the ion exchange fibers. The activity of the obtained immobilized fructosyltransferase is larger than or equal to 180 U/g, the half-life period of the immobilized fructosyltransferase is larger than or equal to 40 times, the saccharose inversion rate is larger than or equal to 90%, and the content of fructo-oligose generated through the saccharose inversion is 55-60%. Compared with the public prior art, the method has the advantages that enzyme extraction is simple, operation time is short, enzymes can be easily recycled, the immobilization process is conducted without cross-linking agents, and the product quality and safety are high.

Description

technical field [0001] The invention relates to a method for immobilizing fructosyltransferase, in particular to a preparation method for immobilizing fructosyltransferase by using ion-exchange fibers, which includes fermentation of mycelium with high activity of fructosyltransferase, enzymatic cell decomposition and extraction of enzyme liquid, Immobilization of fructosyltransferase on ion-exchange fibers. Background technique [0002] Ion Exchange Fiber (IEF for short) is a fibrous ion exchange material. As a new functional polymer material, ion exchange fiber has unique chemical and physical adsorption and separation functions, and plays an irreplaceable role in some related fields. . Ion exchange fibers have the following characteristics: various shapes, can be made into long and short fibers, can also be woven and braided, easy to make various components, and can choose the best shape according to the application purpose, large exchange capacity, specific surface ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02C12N9/10C12R1/69
Inventor 方昱
Owner LINYI CITY HONGYU BIOTECH CO LTD
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