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Method for preparing inositol through enzymic catalysis and inositol

A technology for catalyzing the preparation of inositol, which is applied in the direction of fermentation, can solve the problems of complex components of cell fermentation liquid, low conversion rate of inositol, complex purification of inositol, etc., and achieve the effect of low production cost, high purity and simple production process

Inactive Publication Date: 2016-09-07
浙江欣诺医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in microbial cells, glucose participates in multiple metabolic pathways to maintain cell growth. In addition, inositol-1-phosphate synthase catalyzes glucose-6-phosphate to generate inositol-1-phosphate, and glucose-6-phosphate is produced by Obtained by the phosphorylation of glucose, which requires energy consumption (ATP or phosphoenolpyruvate), and the conversion rate of glucose to inositol is low
In addition, due to the complex composition of the cell fermentation broth, the process of purifying inositol from it is also relatively complicated and the cost is high

Method used

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  • Method for preparing inositol through enzymic catalysis and inositol
  • Method for preparing inositol through enzymic catalysis and inositol
  • Method for preparing inositol through enzymic catalysis and inositol

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Expression of inositol-1-phosphate synthase (INO1) in Escherichia coli

[0043] Search for the inositol-1-phosphate synthase (INO1) gene or protein of thermophilic bacteria on NCBI, screen for microorganisms with an optimal growth temperature of 60-90°C, purchase strains to extract genomes and amplify related genes, or directly synthesize them Related genes, taking inositol-1-phosphate synthase (INO1) derived from Archaeoglobusfulgidus as an example, its protein sequence is numbered GeneBank on NCBI: AIG98795.1, and the sequence is handed over to Gene Synthesis Company to synthesize the corresponding DNA sequence, and Add EcoR I-Nde I and Xho I enzyme cutting sites at its two ends respectively. After the synthetic DNA and the E. coli expression vector such as pET-26b(+) were digested and recovered by Nde I and Xho I respectively, they were ligated overnight at 16°C with T4 DNA ligase, and 5 μL of the ligation product was transformed into E. coli DH5α, and the transforma...

Embodiment 2

[0069] The enzyme prepared in Example 1 was used for experiments. The starch substrate is adjusted with water to make starch milk with a dry matter content of 10%, 0.2M sodium dihydrogen phosphate is added, the pH is adjusted to 6.5, and gelatinization is carried out at 65°C. Add the enzyme powder obtained by spray drying, wherein αGP, PGM, INO1, and MIP are mixed in equal proportions according to the enzyme activity unit, and the addition amount is 1000U-10000U / L, and the four enzymes of αGP, PGM, INO1, and MIP are added per liter of reaction system Take 10000U as an example, according to the data of activity determination, at pH 7.0 and 70°C, the enzyme activities are 3U / mg, 275U / mg, 1.7U / mg, 1.7U / mg respectively. That is, the enzyme amounts of the four enzymes in the mixed enzyme are 3333 mg, 36 mg, 5882 mg, and 5882 mg, respectively. Add the mixed enzyme into the starch milk, keep the temperature constant, and react for 72 hours, and the inositol conversion rate can reach...

Embodiment 3

[0071] Adjust the starch raw material with water to make starch milk with a dry matter content of 10%, add 0.1M sodium dihydrogen phosphate, adjust the pH to 7.0, and conduct gelatinization at 75°C. Add the enzyme powder that spray-drying obtains, wherein αGP (derived from Aquifex aeolicus), PGM (derived from Thermococcus kodakaraensis), INO1 (derived from Archaeoglobusfulgidus), MIP (derived from Archaeoglobusfulgidus) add-on amount is 10000U, maintains temperature constant, After reacting for 72 hours, the inositol content was 67g / L, according to the starch molecular formula (C 6 h 10 o 5 )n, if 100g / L of starch is completely converted into inositol, 111g / L of inositol can be obtained, and the conversion rate of inositol can be calculated as 62÷111×100%=60%.

[0072] αGP and PGM were derived from Thermus thermophilus, INO1 was derived from Archaeoglobusfulgidus, MIP was derived from Thermotoga maritima, and the conversion rate was 62%. Replacing MIP with a Thermus thermop...

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Abstract

The invention relates to a method for preparing inositol through enzymic catalysis. The method comprises the following steps: performing enzyme-catalyzed conversion on a polysaccharide or an oligosaccharide which takes glucose as a unit to obtain glucose-1-phosphate; performing enzyme-catalyzed conversion on the glucose-1-phosphate to obtain glucose-6-phosphate; performing enzyme-catalyzed conversion on the glucose-6-phosphate to obtain inositol-1-phosphate; performing enzyme-catalyzed conversion on the inositol-1-phosphate to obtain inositol. The invention also provides an inositol product which is prepared according to the method. According to the method, the conversion efficiency of the inositol is effectively improved; moreover, a production process is simple, pollution-free, and low in production cost.

Description

technical field [0001] The invention relates to a method for preparing inositol by enzymatic catalysis and a product thereof. Background technique [0002] Cyclohexanol, also known as inositol, is an essential growth factor for animals and one of the components of cell membranes. In the field of medicine, inositol and its derivatives can promote cell growth, prevent and treat arteriosclerosis, hypercholesterolemia and other diseases. In the field of food, inositol is often used as a nutritional enhancer, which can promote development, has anti-oxidation function, and can be used in combination with carnitine to reduce weight. In 1987, the United States recommended that a certain amount of inositol should be added to milk-containing baby food. In addition, inositol is also used in feed to promote animal growth and prevent hair loss. [0003] At present, the main method of inositol production is to extract calcium phytate from substrates such as rice bran or bran, and then ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/24C12P19/18C12P7/02C12P19/02
CPCC12P19/24C12P7/02C12P19/02C12P19/18
Inventor 刘文山汪小锋汪卫
Owner 浙江欣诺医药有限公司
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