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Method for preparing inositol-1-phosphate

A technology of inositol and phosphoric acid, which is applied in the field of preparation of inositol-1-phosphate, can solve the problems of no industrial value, high price, high production cost, etc., and achieve large-scale production, low production cost and high utilization rate of raw materials Effect

Inactive Publication Date: 2019-04-19
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high production cost and high price of glucose-6-phosphate, the production of inositol-1-phosphate from glucose-6-phosphate has no industrial value

Method used

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  • Method for preparing inositol-1-phosphate

Examples

Experimental program
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experiment example 1

[0036] Experimental example 1 in vitro multi-enzyme catalyzed conversion of starch into inositol-1-phosphate

[0037] Starch was converted into inositol-1-phosphate ( figure 1 ). These key enzymes include: (1) starch phosphorylase (αGP, EC 2.4.1.1), which releases glucose-1-phosphate from starch; (2) glucose phosphomutase (PGM, EC 5.4.2.2), which catalyzes Glucose-1-phosphate generates glucose-6-phosphate; (3) inositol-1-phosphate synthase (IPS, EC 5.5.1.4), which catalyzes the generation of inositol-1-phosphate from glucose-6-phosphate. Since the last enzyme reaction is an irreversible reaction, the conversion rate of the enzyme-catalyzed system is very high.

[0038] In the present invention, starch phosphorylase and glucose phosphate mutase are derived from Thermotoga maritima, and the gene numbers on KEGG are TM1168 and TM0769 respectively, and inositol-1-phosphate synthase is derived from Archaeoglobusfulgidus, and the gene numbers on KEGG For AF1794, these genomic DN...

experiment example 2

[0045] Experimental example 2 in vitro multi-enzyme catalyzed conversion of starch into inositol-1-phosphate (reaction at different temperatures)

[0046] The preparation of starch phosphorylase, glucose phosphomutase and inositol-1-phosphate synthase was the same as in Experimental Example 1.

[0047] In a 1.0mL reaction system containing 200mM sodium phosphate buffer (pH 7.2), 5mM divalent magnesium ions, 0.5U / mL starch phosphorylase, 0.1U / mL glucose phosphomutase, and 0.5 U / mL inositol-1-phosphate synthase, 20g / L soluble starch, catalyzed reaction at 80°C, and the reaction time was 72 hours.

[0048] After the reaction, the final concentration of inositol-1-phosphate was 6.4g / L, and the conversion rate to starch was 32%.

experiment example 3

[0049] Experimental Example 3 Improve the yield of inositol-1-phosphate through process optimization and adding enzymes that promote starch hydrolysis

[0050] Since starch phosphorylase cannot completely hydrolyze starch, the yield of inositol-1-phosphate is limited. Therefore, we added isoamylase (isoamylase, EC 3.2.1.68) that can help starch hydrolysis in the reaction system. Under the double action of starch phosphorylase and isoamylase, the end products of starch hydrolysis are maltose and glucose. In order to convert these products into inositol-1-phosphate, maltose phosphorylase (maltose phosphorylase, EC 2.4.1.8) and polyphosphate glucokinase (polyphosphate glucokinase, EC 2.7.1.63) need to be added.

[0051] In the present invention, the isoamylase is derived from Sulfolobus tokodaii, and the number of the gene on KEGG is ST0928, and the genome DNA of this bacterial strain is German Albert-Ludwigs- Courtesy of Prof. Georg Fuchs, Freiburg. Polyphosphoglucokinase is...

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Abstract

The invention discloses a method for preparing inositol-1-phosphate, which belongs to the field of enzyme catalysis preparation of inositol-1-phosphate. The preparation method of the inositol-1-phosphate disclosed by the invention uses starch and its derivative and phosphate as substrates to efficiently convert the substrate into inositol-1-phosphate by in vitro multi-enzyme molecular machine catalysis. The method adds the enzyme capable of promoting hydrolysis of starch and its derivative and the enzyme utilizing by-product glucose to obviously increase the yield of the inositol-1-phosphate.The method of the invention has high yield of inositol-1-phosphate, simple production equipment, and low production cost, and can realize large-scale prepration of inositol-1-phosphate.

Description

technical field [0001] The invention relates to a method for preparing inositol-1-phosphate, in particular to a method for converting starch and its derivatives into inositol-1-phosphate by catalyzing an in vitro multi-enzyme molecular machine, and belongs to the field of enzyme-catalyzed production of inositol-1-phosphate . Background technique [0002] Inositol is one of the water-soluble vitamin B family, an essential substance for the growth of humans, animals and plants, and is widely used in medicine, food, feed and other industries. Inositol can promote cell metabolism and improve cell nutrition, so it can promote development, increase appetite and restore physical strength. And it can prevent the accumulation of fat in the liver, accelerate the removal of excess fat in the heart, and has a synergistic fat-dispelling effect with choline, so it is used to treat hyperlipidemia and cirrhosis. In addition, inositol can also prevent animal hair from falling out. Inosito...

Claims

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Application Information

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IPC IPC(8): C12P9/00
CPCC12P9/00
Inventor 张以恒李运杰
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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