Preparation method of glucose 1-phosphoric acid

A technology of glucose and phosphoric acid, applied in the field of preparation of glucose 1-phosphate, can solve the problems of high separation cost, low product yield, different glucan phosphorylase, etc., and achieves reduced separation cost, high raw material utilization, conversion Efficiency-enhancing effect

Inactive Publication Date: 2017-06-09
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

However, the method of simply using polysaccharide phosphorylase to catalyze the production of glucose 1-phosphate in the past has many shortcomings
For example, when disaccharides (sucrose, trehalose or cellobiose) are used as substrates, the price of disaccharides may be much higher than that of starch. Taking sucrose as an example, the theoretical maximum of glucose 1-phosphate calculated by glucose is The production rate is only 50%, and the production process will be accompanied by the production of fructose, resulting in high separation costs
Although the cost of maltodextrin and starch is lower than that of disaccharides, the presence of α-1,6 glycosidic bonds in the substrate causes glucan phosphorylase to not fully react with the substrate, and the substrate of glucan phosphorylase Choose maltotetraose or longer oligosaccharides, maltose and glucose cannot be used as substrates for glucan phosphorylase, these reasons will lead to low product yield when maltodextrin or starch is used as a substrate

Method used

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Examples

Experimental program
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Effect test

experiment example 1

[0044] Experimental example 1 Multienzyme Catalyzed Conversion of Starch to Glucose in Vitro 1- Phosphoric acid, improves starch conversion rate

[0045] Starch is converted to glucose 1-phosphate by an in vitro multi-enzyme catalytic system ( figure 1 ). These key enzymes include: (1) branching enzyme IA; (2) glucan phosphorylase (αGP, EC 2.4.1.1), which releases glucose-1-phosphate from starch; (3) glucantransferase ( 4-a-glucanotransferase, EC 2.4.1.25).

[0046] Since starch has branched chains, starch cannot be completely hydrolyzed by glucan phosphorylase alone, because glucan phosphorylase only acts on α-1, 4 glycosidic bonds, and branched chains are based on α-1, 6 glycosidic bonds connected to the main chain. This requires the addition of isoamylase (isoamylase, EC 3.2.1.68) to hydrolyze the α-1,6 glycosidic bond. Since the minimum substrate of glucan phosphorylase is maltotriose, in order to improve the utilization efficiency of maltodextrin, glucan tran...

experiment example 2

[0052] Experimental example 2 Multienzyme Catalyzed Conversion of Starch to Glucose in Vitro 1- phosphoric acid

[0053] The preparation of isoamylase, glucan phosphorylase, and glucan transferase is the same as in Experimental Example 1.

[0054] In a 0.75 mL reaction system containing 200 mM phosphate buffer (pH 7.2), 5 mM divalent magnesium ion, 0.05 U / mL dextran phosphorylase, 1 U / mL isoamylase, and 1 U / mL of glucan transferase and 10 g / L of soluble starch were catalyzed at 40°C for 40 hours.

[0055] After the reaction, the final concentration of glucose 1-phosphate was 2.2 g / L, and the conversion rate was 22%.

experiment example 3

[0056] Experimental example 3 Multienzyme Catalyzed Conversion of Starch to Glucose in Vitro 1- phosphoric acid

[0057] The preparation of isoamylase, glucan phosphorylase, and glucan transferase is the same as in Experimental Example 1.

[0058] In a 0.75 mL reaction system containing 200 mM phosphate buffer (pH 7.2), 5 mM divalent magnesium ion, 0.05 U / mL dextran phosphorylase, 1 U / mL isoamylase, and 1 U / mL of glucan transferase and 10 g / L of soluble starch were used to catalyze the reaction at 80°C for 40 hours.

[0059] After the reaction, the final concentration of glucose 1-phosphate was 12.1 g / L, and the conversion rate was 121%.

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Abstract

The invention discloses a preparation method of glucose 1-phosphoric acid, belonging to the technical field of enzyme catalysis of glucose 1-phosphoric acid. According to the preparation method of glucose 1-phosphoric acid, starch or a starch derivative is taken as a substrate and is converted into glucose 1-phosphoric acid through efficient catalysis of acellular multienzyme in a multienzyme reaction system. By optimizing the process, enzyme capable of promoting starch hydrolysis and enzyme utilizing a side product maltose are added so as to establish the multienzyme reaction system, so that the conversion efficiency of the raw materials is remarkably improved, and the yield of glucose 1-phosphoric acid is remarkably increased. The preparation method has the beneficial effects that the conversion rate of the raw materials is high, the yield of glucose 1-phosphoric acid is high, the steps are simple, the production cost is low, the environment is hardly affected, and the large-scale production of glucose 1-phosphoric acid can be realized.

Description

technical field [0001] The invention relates to a method for preparing glucose 1-phosphate, in particular to a method for converting starch and starch derivatives into glucose 1-phosphate by cell-free multi-enzyme catalysis, and belongs to the field of enzyme-catalyzed production of glucose 1-phosphate. Background technique [0002] In organisms, glucose 1-phosphate (G 1-P) is produced by glucan phosphorylase catalyzing the reaction of polysaccharides such as glycogen, starch or maltodextrin and inorganic phosphate, which is the first step in the gluconeogenesis pathway A metabolite that can generate glucose-6-phosphate under the action of phosphoglucomutase (PGM, EC 5.4.2.2) and enter glycolysis or other metabolic pathways. As an activated glucose, G 1-P is an important precursor in the synthesis of complex carbohydrate compounds (such as glycolipids, oligosaccharides, sugar nucleotides). In addition, the compound can also be used to synthesize artificial starch, generate ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/02C12P19/14C12P19/18
Inventor 张以恒周伟
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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