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Production method of L-erythrulose and erythritol

A technology of erythrulose and erythritol, which is applied in the production field of L-erythrulose and erythritol, and can solve the problems of long fermentation cycle, high cost of raw materials, and difficulty in increasing product scale

Active Publication Date: 2019-07-05
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

[0004] In the existing erythritol production technology, the raw material glucose comes from the hydrolysis of food crops, and large-scale production will pose a threat to food security
In addition, the long conversion pathway of glucose to erythritol in the fermenting microorganisms results in long fermentation cycles, increased costs and the risk of bacterial contamination
However, the production of L-erythrulose uses erythritol as raw material, the cost of raw materials is high and limited by the production capacity of erythritol, it is difficult to increase the product scale

Method used

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  • Production method of L-erythrulose and erythritol
  • Production method of L-erythrulose and erythritol

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Embodiment 1

[0144] Embodiment 1, preparation and functional detection of enzymes used in the production of L-erythrulose and erythritol

[0145] figure 1 Schematic diagram of the new reaction catalyzed by tktA. figure 2 Schematic diagram of the new reaction catalyzed by GoXDH. image 3 Schematic diagram of the reaction for the new method of producing erythritol using glycolaldehyde. Figure 4 It is a schematic diagram of the reaction of the new method for producing L-erythrulose by utilizing ethylene glycol.

[0146] 1. Preparation of enzymes tktA, GoXDH, EgDH, NOX2 and FDH required for the production of L-erythrulose and erythritol 1. Acquisition of DNA encoding enzymes

[0147] tktA, the amino acid sequence is sequence 1, from Escherichia coli (Escherichia coli) K12, the nucleotide sequence of the DNA molecule encoding the protein is sequence 6;

[0148] GoXDH, the amino acid sequence is sequence 2, from Gluconobacter oxydans; under the premise of not changing the amino acid sequen...

Embodiment 2

[0196] The production method of embodiment 2, L-erythrulose and erythritol

[0197] 1. Production of L-erythrulose with glycolaldehyde as raw material

[0198] A. tktA enzyme catalyzes the production of L-erythrulose from glycolaldehyde

[0199] 1. Prepare 50mM potassium phosphate buffer (adjust pH to 6.5, 7.0, 7.5, 8.0 respectively), add 1mM TPP and 5mM magnesium sulfate, add 0.5mg / mL tktA enzyme, preheat at 37°C for 5min, add raw material 200mM hydroxyethyl Aldehydes start to react. React at 37°C for 30 minutes. 100 μL of the reaction product was added to 100 μL of acetonitrile, mixed thoroughly, and then centrifuged at 12,000 rpm for 30 minutes, and 100 μL of the supernatant was taken for HPLC analysis. The result is as Figure 21 As shown, the yield of L-erythrulose was the highest when the pH of the buffer solution was 7.

[0200]2. Prepare 50mM potassium phosphate buffer (pH=7.0), add 1mM TPP and different concentrations of magnesium sulfate (0, 0.01, 0.1, 1, 5, 10m...

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Abstract

The invention discloses a production method of L-erythrulose and erythritol. The invention provides any one of 1) to 3) applications: 1) an application of tktA enzyme or related biological materials thereof in the production of L-erythrulose, wherein the substrate of the application does not contain hydroxypyruvic acid; 2) an application of GoXDH enzyme or related biological materials thereof in the production of erythritol; and 3) an application of the tktA enzyme or related biological materials thereof and the GoXDH enzyme or related biological materials thereof in the production of erythritol, wherein the substrate of the application does not contain hydroxypyruvic acid. In summary, the present invention provides a new approach for converting glycolaldehyde to L-erythrulose or L-erythritol.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for producing L-erythrulose and erythritol. Background technique [0002] L-erythrulose (L-(+)-Erythrulose) is a rare sugar that can be used as a starting material for food additives, cosmetic raw materials, pharmaceutical precursor compounds, and other rare sugars. Large, high value in industrial applications. Erythritol is a four-carbon polyol that can be used as a nutritional sweetener in food and other fields. Erythritol has the advantages of low energy value, high tolerance, no side effects, edible for diabetics, non-cariogenic, etc., and is suitable for use in functional foods. Erythritol may replace xylitol in the future. [0003] At present, the production of L-erythrulose mainly uses erythritol as raw material, which is obtained by chemical method or biological fermentation method. At present, the main production method of erythritol is microbial fermentation me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/02C12P7/18C12N15/70C12N15/81C12R1/19
CPCC12P19/02C12P7/18
Inventor 江会锋丁文涛刘玉万杨一群
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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