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Technique for developing allulose-containing functional jujube juice through biotransformation of jujube monosaccharide

A technique of psicose and biotransformation, which is applied in the application of coupled immobilized double enzymes in the production of psicose from concentrated red jujube juice, site-directed mutation of glucose isomerase and D-psicose 3-epimer In the field of co-expression and preparation of constitutive enzymes, it can achieve the effects of reducing environmental pollution, shortening the reaction process and reducing the cost of separation.

Inactive Publication Date: 2014-05-14
SHANXI TIANJIAO FOOD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no publication of the co-expression of glucose isomerase and D-psicose 3-epimerase to obtain the two enzymes at the same time, and there is no disclosure of the site-directed mutation of the 146-amino acid of glucose isomerase

Method used

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  • Technique for developing allulose-containing functional jujube juice through biotransformation of jujube monosaccharide
  • Technique for developing allulose-containing functional jujube juice through biotransformation of jujube monosaccharide
  • Technique for developing allulose-containing functional jujube juice through biotransformation of jujube monosaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] A method for producing psicose from jujube juice using double-enzyme coupling, immobilization and catalytic jujube juice is carried out according to the following steps:

[0111] first step, immobilization

[0112] Add the crude enzyme solution to bisphenol A epoxy vinyl ester resin, add 2ml of crude enzyme solution per 1.0g of resin, the adsorption temperature is 35°C, the adsorption pH=7.5, the adsorption time is 140min, and then add the cross-linking agent pentadiene Aldehyde, crosslinking time 35min, described crosslinking agent volume fraction is 0.06%, obtains immobilized enzyme;

[0113] In the second step, double-enzyme coupling catalyzes the production of allulose from jujube juice

[0114] Add jujube juice to the immobilized enzyme, control the temperature: 70°C; pH: 7.0; the weight ratio of the jujube juice to the immobilized enzyme is 2:1; complete the catalytic reaction in 10 hours to obtain allulose Mixture.

Embodiment 2

[0116] A method for producing psicose from jujube juice using double-enzyme coupling, immobilization and catalytic jujube juice is carried out according to the following steps:

[0117] first step, immobilization

[0118] Add the crude enzyme liquid to the bisphenol A epoxy vinyl ester resin, add 0.5ml of the enzyme amount per 1.0g of the resin, the ratio of the two enzymes in the crude enzyme liquid is 4:1, the adsorption temperature is 25°C, and the adsorption temperature is 25°C. pH 6.5, adsorption time 100min, then add crosslinking agent glutaraldehyde, crosslinking time 25min, the volume fraction of the crosslinking agent is 0.03%, to obtain immobilized enzyme;

[0119] In the second step, double-enzyme coupling catalyzes the production of allulose from jujube juice

[0120] Add jujube juice to the immobilized enzyme, control the temperature: 60°C; pH: 5.0; the weight ratio of the jujube juice to the immobilized enzyme is 1:1; complete the catalytic reaction in 6 hours t...

Embodiment 3

[0122] A method for producing psicose from jujube juice using double-enzyme coupling, immobilization and catalytic jujube juice is carried out according to the following steps:

[0123] first step, immobilization

[0124] Add the crude enzyme solution to bisphenol A epoxy vinyl ester resin, add 1ml of crude enzyme solution per 1.0g of resin, the adsorption temperature is 30°C, the adsorption pH=7, the adsorption time is 120min, and then add the cross-linking agent pentadiene Aldehyde, crosslinking time 30min, described crosslinking agent volume fraction is 0.05%, obtains immobilized enzyme;

[0125] The second step is to catalyze the production of allulose from jujube juice

[0126] Add jujube juice to the immobilized enzyme, control the temperature: 65°C; pH value: 6.0; the weight ratio of the jujube juice to the immobilized enzyme is 3:1; the catalytic reaction is completed in 8 hours to obtain allulose Mixture.

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Abstract

The invention relates to a two-enzyme co-expression technique and a two-enzyme coupling and immobilizing technique, and specifically relates to a co-expression preparation method of a site-specific mutant glucose isomerase and a D-allulose 3-epimerase, a coupling and immobilizing technique of the two enzymes and application of the coupled and immobilized two enzymes in producing allulose from jujube condensed juice. In order to solve health problems of people due to high-calorific value sugar, the invention provides a method for promoting the production of low-calorific value D-allulose from the recombinase glucose isomerase and the D-allulose 3-epimerase through co-immobilization and two-enzyme coupling by virtue of a multiple-enzyme coupling and immobilizing technique with D-glucose and D-fructose in the jujube condensed juice as the substrates, and also provides a method for site-specific mutagenesis of an amino acid at the 146th site of the glucose isomerase and co-expression thereof with the D-allulose 3-epimerase.

Description

technical field [0001] The present invention relates to double-enzyme co-expression technology and double-enzyme coupling and immobilization technology, in particular to a preparation method for the co-expression of site-directed mutation glucose isomerase and D-psicose 3-epimerase and the double-enzyme The coupling and immobilization technology and the application of coupling and immobilization double enzymes in the production of psicose from jujube concentrated juice. Background technique [0002] Jujube integrates nutrition, health care and medicinal functions, and is listed in the first batch of medicinal and edible fruits of the Ministry of Health. Jujube is rich in sugar substances, and its sugar content ranks first among all kinds of fruits, higher than that of sugarcane and sugar beet, which are raw materials for sugar making. The sugar content of fresh dates is 20%-36%, and the sugar content of dried dates is as high as 50-80%. The sugars in dates can be divided in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/24C12P19/02C12N11/18C12N9/92C12N9/90C12N15/70C12R1/19C12R1/07C12R1/01
Inventor 王晓艳门燕冯俊敏孙媛霞张佩舜朱玥明康振奎巩晋龙郑丽萍王小鹏李奠础
Owner SHANXI TIANJIAO FOOD
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