Unlock instant, AI-driven research and patent intelligence for your innovation.

A kind of preparation method of L-type rare ketohexose

A kind of rare ketohexose, L-type technology, applied in the field of preparation of L-type rare ketohexose, can solve the problems of difficult separation and purification of products, low conversion rate, etc., to reduce the use of NAD+, low production cost, and cost reduction Effect

Active Publication Date: 2022-05-03
JIANGNAN UNIV
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the thermodynamic equilibrium involved, the conversion rate is low, and the product is difficult to separate and purify

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of preparation method of L-type rare ketohexose
  • A kind of preparation method of L-type rare ketohexose
  • A kind of preparation method of L-type rare ketohexose

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0046]The preparation method of L-fructose of the present invention comprises the following steps: (1) using glycerol and pyrophosphate as substrates, adding L-rhamnulose-1-phosphate aldolase (L-rhamnulose-1-phosphate aldolase, EC 4.1.2.19), glycerol phosphate oxidase (L-α-glycerophosphate oxidase, EC: 1.1.3.21), catalase (Catalase, EC: 1.11.1.6), acid phosphatase (acid phosphatase, EC: 3.1. 3.2) Multi-enzyme catalysts of horse liver alcohol dehydrogenase (horse livercohol dehydrogenase, EC: 1.1.1.2), NADH oxidase (NADH oxidase, EC: 1.6.3.4) to establish a multi-enzyme reaction system for enzymatic reactions; (2 ) to separate and purify the enzyme-catalyzed reaction product.

[0047] Wherein, the glycerol concentration in step (1) is 300-900mM; preferably, the glycerol concentration is 500-800mM, most preferably 800mM; the pyrophosphate concentration is 5-125mM, preferably, the pyrophosphate The concentration is 20-100mM; the most preferred is 40mM; wherein the pyrophosphate ...

experiment example 1

[0050] Experimental example 1: In vitro multi-enzyme catalyzed conversion of glycerol to L-fructose

[0051] Horse liver alcohol dehydrogenase can specifically oxidize glycerol to generate L-glyceraldehyde, without producing other enantiomers, and can be used for L-glyceraldehyde production. However, NAD is required for its production + , the production cost is higher. NADH oxidase can transfer electrons from NADH to oxygen to generate H2O. This study is the first to combine horse liver alcohol dehydrogenase and NADH oxidase at low concentrations of NAD + L-GA is produced under the action.

[0052] The 0.5mL reaction system contains 50mM Tris-Hcl (pH7.5), the dosage of horse liver alcohol dehydrogenase is 16U / mL, the dosage of NADH oxidase is 32U / mL, glycerol 100mM and NAD + 1 mM; another reaction without NADH oxidase served as a control. The amount of D-glyceraldehyde in the reaction solution was measured by HPLC at 6h of reaction. The result is as figure 1 As shown, t...

experiment example 2

[0057] Experimental example 2: In vitro multi-enzyme catalyzed conversion of glycerol into L-fructose and L-tagatose

[0058] Conversion of glycerol to L-fructose and L-tagatose ( Image 6 ). These key enzymes include: (1) acid phosphatase (PhoN-Sf, EC: 3.1.3.2), which phosphorylates glycerol to generate DL-3-phosphate glycerol. At the same time, sugar phosphate is dephosphorylated to generate rare ketohexose; (2) glycerol phosphate oxidase (GPO, EC: 1.1.3.21), which catalyzes DL-3-glycerol phosphate to dihydroxyacetone phosphate (DHAP); (3) liver Alcohol dehydrogenase (horse l iver alcohol dehydrogenase, EC: 1.1.1.2), converts glycerol into L-glyceraldehyde; (4) L-fucose-1-phosphate aldolase (FucA, EC 4.1.2.17 ), will convert DHAP and L-glyceraldehyde into sugar phosphate; (5) catalase (Catalase, EC: 1.11.1.6), decompose hydrogen peroxide into water and oxygen; (6) NADH oxidase (NADH oxidase , EC:1.6.3.4), convert NADH to NAD + .

[0059] L-fucose-1-phosphate aldolase de...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a preparation method of L-type rare ketohexose, which comprises: using glycerol and pyrophosphate as substrates, adding L-rhamnosan-1-phosphate aldolase, glycerol phosphate oxidase, peroxide Catalase, acid phosphatase, horse liver alcohol dehydrogenase, and NADH oxidase establish a multi-enzyme reaction system for enzymatic reactions; the enzyme-catalyzed reaction products are separated and purified. The raw material price of the present invention is low, does not need glycerol kinase, does not need ATP to carry out substrate phosphorylation to produce glycerol triphosphate, greatly reduces the cost of producing L-type rare ketohexose, and the present invention combines horse liver alcohol dehydrogenase and NADH oxidase Produce L-glyceraldehyde, reduce NAD + usage of.

Description

technical field [0001] The invention belongs to the technical field of enzyme-catalyzed preparation of L-type rare ketohexose, and in particular relates to a preparation method of L-type rare ketohexose. Background technique [0002] Rare ketohexoses are a class of monosaccharides and their derivatives that exist in nature but at very low levels and have important physiological functions. The L-configuration includes L-sorbose, L-fructose and L-tagatose, etc. It has the advantages of low calorie and natural sweetness, and can be used as a functional sweetener. It also has anti-cancer, anti-obesity, neuroprotection, Physiological activities such as scavenging free radicals have broad application prospects in the fields of life medicine, cosmetics, and diet. Therefore, it is of great significance to improve the rare sugar synthesis system. [0003] There are two methods for producing L-type rare ketohexose, chemical synthesis and biotransformation. The traditional chemical ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/02
CPCC12P19/02
Inventor 李子杰高晓冬李芬
Owner JIANGNAN UNIV