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Method for synthesizing soybean oligosaccharides through biological catalysis

A technology of soybean oligosaccharide and synthase, applied in the direction of fermentation, can solve the problems of high price, difficulty and low yield of plant extraction method.

Inactive Publication Date: 2018-01-16
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the source of commercial soybean oligosaccharides is the plant extraction method, which is directly extracted from soybean seeds. However, the extraction rate of the plant extraction method is low, and it is difficult to obtain a single soybean oligosaccharide with high purity, resulting in pure inositol The price of galactoside, raffinose or stachyose is very high. Therefore, it is urgent to develop a new method for preparing high-purity soybean oligosaccharides with low cost and low pollution.

Method used

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  • Method for synthesizing soybean oligosaccharides through biological catalysis
  • Method for synthesizing soybean oligosaccharides through biological catalysis
  • Method for synthesizing soybean oligosaccharides through biological catalysis

Examples

Experimental program
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Effect test

Embodiment 1

[0051] Example 1. In vitro multi-enzyme catalyzed conversion of sucrose into galactoside

[0052] The conversion of sucrose to galactoside ( figure 1 ). These key enzymes include: (1) sucrose synthase (SS, EC 2.4.1.13), which catalyzes the synthesis of UDP-glucose from sucrose and uridine diphosphate (UDP); (2) uridine diphosphate glucose 4-epimerase (UGE, EC 5.1.3.2), which catalyzes the synthesis of UDP-galactose from UDP-glucose; (3) galactoside synthase (GS, EC 2.4.1.123), which catalyzes the synthesis of inositol hemi-inositol from UDP-galactose and inositol lactose.

[0053] In the present invention, sucrose synthase and galactinol synthase are derived from Arabidopsisthaliana, the gene numbers on KEGG are AT5G20830 and AT1G09350 respectively, and uridine diphosphate glucose 4-epimerase is derived from large intestine The gene number of Escherichia coli on GeneBank is 945354, and these genomic DNAs can be obtained from the official website of ATCC (www.atcc.org). The...

Embodiment 2

[0061] Example 2: In vitro multi-enzyme catalyzed conversion of sucrose into raffinose

[0062] Establishment of an in vitro multi-enzyme catalytic system to convert sucrose into raffinose ( figure 1 ). These key enzymes include: (1) sucrose synthase (SS, EC 2.4.1.13), which catalyzes the synthesis of UDP-glucose from sucrose and uridine diphosphate (UDP); (2) uridine diphosphate glucose 4-epimerase (UGE, EC 5.1.3.2), which catalyzes the synthesis of UDP-galactose from UDP-glucose; (3) galactoside synthase (GS, EC 2.4.1.123), which catalyzes the synthesis of inositol hemi-inositol from UDP-galactose and inositol Lactose; (4) Raffinose synthase (RS, EC 2.4.1.82), which catalyzes the synthesis of raffinose and inositol from galactositol and sucrose.

[0063] The sources of sucrose synthase, galactinol synthase, and uridine diphosphate glucose 4-epimerase are the same as in Example 1. The raffinose synthase is derived from Arabidopsis thaliana, and the gene is on KEGG The seri...

Embodiment 3

[0068] Example 3, in vitro multi-enzyme catalyzed conversion of sucrose into stachyose

[0069] An in vitro multi-enzyme catalytic system was established to convert sucrose into stachyose ( figure 1 ). These key enzymes include: (1) sucrose synthase (SS, EC 2.4.1.13), which catalyzes the synthesis of UDP-glucose from sucrose and uridine diphosphate (UDP); (2) uridine diphosphate glucose 4-epimerase (UGE, EC 5.1.3.2), which catalyzes the synthesis of UDP-galactose from UDP-glucose; (3) galactoside synthase (GS, EC 2.4.1.123), which catalyzes the synthesis of inositol hemi-inositol from UDP-galactose and inositol Lactose; (4) raffinose synthase (RS, EC 2.4.1.82), which catalyzes the synthesis of raffinose and inositol from galactose inositol and sucrose; (5) stachyose synthase (SS, EC: 2.4.1.67 ), catalyzing the synthesis of stachyose and inositol from galactoside and raffinose.

[0070] The sources of sucrose synthase, galactinol synthase, uridine diphosphate glucose 4-epime...

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Abstract

The invention discloses a method for synthesizing soybean oligosaccharides through biological catalysis. The method has the beneficial effects that that through establishing an in-vitro multi-enzyme cascade reaction system composed of sucrose synthetase, uridine diphosphate glucose 4-differential isomerase, inositol galactoside synthase, raffinose synthase and threonose synthase, sucrose is takenas a substrate to synthesize a single component or a mixed component in inositol galactoside, raffinose and threonose; the method realizes the recycling conversion of uridine diphosphate and inositol,and a crude enzyme solution added in the system contains a small amount of uridine diphosphate, so that the expensive uridine diphosphate can be avoided; the inositol galactoside yield reaches 50 g / L, the conversion rate is 70 percent, the raffinose yield reaches 20 g / L, the threonose yield reaches 10 g / L, and the obtained inositol galactoside, raffinose and threonose can be used in the fields offood, medicines and the like; the method disclosed by the invention is of great significance in the utilization of high attachment values of the sucrose and a biomass rich in the sucrose.

Description

technical field [0001] The invention relates to the field of biocatalysis, in particular to an in vitro multi-enzyme cascade catalyzed method for converting sucrose into soybean oligosaccharides. Background technique [0002] Soy oligosaccharides are the general term for soluble sugars in soybeans. The main ingredients refer to sucrose (disaccharides), raffinose (trisaccharides) and stachyose (tetrasaccharides) with a monosaccharide number of 3 to 4. It is a sweetener with low sweetness and low calories. Replace sucrose in functional food or low-energy food; improve the balance of intestinal flora, prevent constipation; inhibit the production of harmful substances in the intestinal tract; reduce the concentration of serum cholesterol and blood lipids; enhance the body's immunity and inhibit the growth of tumor cells; Promote the generation and absorption of nutrients in the intestines and protect the liver. Soybean oligosaccharides are fermented or partially fermented in t...

Claims

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Application Information

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IPC IPC(8): C12P19/24C12P19/14C12P19/12C12P19/00
Inventor 曾燕杨建刚戴隆海张颖孙媛霞
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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