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Method for catalyzing and synthesizing uridine phosphinylidyne compound in an oriented way

A uridine phosphoryl compound, uridine technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., to achieve the effects of low production cost, enhanced enzyme activity, and simple reaction system

Inactive Publication Date: 2009-10-14
NANJING UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are currently no domestic reports on the biocatalytic synthesis of UTP and UDPG from uridine or orotic acid, nor have there been reports on the directional catalytic synthesis of the above three uridine phosphoryl compounds using the same reaction system by fine-tuning the reaction conditions.

Method used

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  • Method for catalyzing and synthesizing uridine phosphinylidyne compound in an oriented way
  • Method for catalyzing and synthesizing uridine phosphinylidyne compound in an oriented way
  • Method for catalyzing and synthesizing uridine phosphinylidyne compound in an oriented way

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Directed production of UMP using uridine.

[0046] In a reaction tank with a capacity of 15L, it is composed of 600mMol of uridine, 5Mol of glucose, 50mMol of magnesium sulfate, 2400g of Brevibacterium ammoniagenes, 20mMol of ammonium chloride, 300mMol of potassium chloride, 2.0Mol of sodium dihydrogen phosphate, 50ml of toluene and water. 10L of the reaction solution, the pH was adjusted to 7.0 with sodium hydroxide, the dissolved oxygen was controlled at 70%, and the temperature was 30°C. After 8 hours of reaction, the reaction was terminated, and the product was precipitated with perchloric acid, and the product was quantitatively analyzed by HPLC. The main product is UMP with a content of 457mMol (208g) and a yield of 76.1%. At this time, the content of UTP is 68.3mMol (33.1g) and the content of UDPG is 24.6mMol (15g).

Embodiment 2

[0047] Example 2: Directed production of UTP using uridine.

[0048] In a reaction tank with a capacity of 15L, it is composed of 600mMol of uridine, 5Mol of glucose, 50mMol of magnesium sulfate, 2400g of Brevibacterium ammoniagenes, 20mMol of ammonium chloride, 300mMol of potassium chloride, 2.0Mol of sodium dihydrogen phosphate, 50ml of toluene and water. 10L of the reaction solution, adjust the pH to 7.0 with sodium hydroxide, control the dissolved oxygen at 70%, and the temperature at 30°C. After 8 hours of reaction, adjust the dissolved oxygen to 25%, adjust the temperature to 37°C, continue the reaction for 6 hours, and end Reaction, precipitation with perchloric acid, carry out quantitative analysis to product with HPLC, main product is UTP in the conversion liquid, and its content is 398mMol (193g), and yield is 66.3%, and UMP content is now 57.3mMol (18.6g), The UDPG content was 80.2 mMol (48.9 g).

Embodiment 3

[0049] Example 3: Directed production of UDPG using uridine.

[0050] In a reaction tank with a capacity of 15L, it is composed of 600mMol of uridine, 5Mol of glucose, 50mMol of magnesium sulfate, 2400g of Brevibacterium ammoniagenes, 20mMol of ammonium chloride, 300mMol of potassium chloride, 2.0Mol of sodium dihydrogen phosphate, 50ml of toluene and water. 10L of the reaction solution, the pH was adjusted to 7.0 with sodium hydroxide, the dissolved oxygen was controlled at 70%, and the temperature was 30°C. After 8 hours of reaction, the dissolved oxygen was adjusted to 25%, and the temperature was adjusted to 37°C. After continuing the reaction for 6 hours, Dissolved oxygen is adjusted to 5%, temperature is adjusted to 25 ℃, continue to react for 4 hours, finish reaction, precipitate with perchloric acid, carry out quantitative analysis to product with HPLC, main product is UDPG in the conversion liquid, and its content is 361mMol (220g) , the yield was 60.2%. At this time,...

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Abstract

The invention discloses a method for catalyzing and synthesizing uridine phosphinylidyne compound in an oriented way; the method takes uridine or saratin and phosphate radical ion as substrate as well as glucose as ribose radical donor and energy donor, utilizes microbial cells with permeability as enzyme source, and promotes the reaction system to biologically catalyzing and synthesizing the uridine phosphinylidyne compound in an oriented way by means of changing reaction temperature and dissolved oxygen periodically. According to the demand, the method can be used for producing any of three matters, and the reaction system is simple, nontoxic, production cost is low and control method is easy, so that the production control in industrialization is facilitated. When the uridine is used as substrate, the yields of UMP, UTP and UDPG are respectively 77%, 67% and 61%; when the saratin is used as substrate, the yields of UMP, UTP and UDPG are respectively 70%, 61% and 53%.

Description

technical field [0001] The invention belongs to the technical field of biocatalysis, and in particular relates to a method for respectively directional catalytically synthesizing uridine phosphoryl compounds by using regulation technology. Background technique [0002] As a derivative of uracil, uridine acid (UMP) is an important biochemical reagent, and it is also a large amount of pharmaceutical intermediates, baby food additives and feed additives, and also such as uridine triphosphate (UTP), urine diphosphate Glucose (UDPG), cytidine triphosphate (CTP), citicoline (CDPC) and other synthetic precursors of uridine and cytidine phosphoryl compounds, a large number of pharmacological and clinical studies have shown that such compounds play a role in the brain It has good clinical effects in the treatment and adjuvant therapy of difficult diseases such as blood vessels, hepatitis, diabetes, anti-virus (SARS) and neurodegeneration (Parkinson's disease, Alzheimer's disease). It...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/30C12R1/185C12R1/265C12R1/72C12R1/84C12R1/78C12R1/645C12R1/01
Inventor 应汉杰杨鹏陈勇张磊黄小权熊健柏建新
Owner NANJING UNIV OF TECH
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