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Method for preparing uridine diphosphate

A technology of uridine diphosphate and uridine triphosphate, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as increased cost, low recovery rate, increased production cost, etc., and achieve shortened time. , The effect of improving product recovery rate and reducing burden

Inactive Publication Date: 2010-05-12
北京燕京中科生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] 1. The concentration of fermentation substrate is low, generally lower than 2% or 20mg / ml, and the amount of product obtained is not high, which leads to low utilization rate of fermentation equipment, low labor productivity, and increased production cost
[0004] 2. The pretreatment process of the fermentation broth is unreasonable, and the method of centrifugation and alcohol precipitation and redissolution is used, which consumes a lot of energy, takes a long time, and the recovery rate is low
[0005] 3. The separation and purification process is unreasonable, and the methods and equipment used in the separation and purification are outdated, which makes the cycle long, the recovery rate is low, the product quality is low, and the cost increases
[0008] But so far, there are still no high-efficiency, low-cost, and easy-to-operate UTP and UDP production processes in China

Method used

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  • Method for preparing uridine diphosphate
  • Method for preparing uridine diphosphate
  • Method for preparing uridine diphosphate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] figure 1 It is the production process flow of uridine diphosphate of the present invention; this embodiment adopts the method of primary fermentation and degradation to produce uridine diphosphate.

[0061] 1. Fermentation

[0062] First, adopt one-step method to ferment uridine monophosphate (6-liter scale), fermentation conditions: UMP318.7g (feeding according to electrophoretic purity 75.3%); Saccharomyces cerevisiae 3kg; Glu 432g; NaH 2 PO 4 280.8g; MgCl 2 24.4g; pH 6.7; H 2 Add O to 6 liters; keep warm at 37°C, and ferment statically for 6 hours.

[0063] Brewer's yeast is eliminated yeast in beer production, and the survival rate of yeast cells is more than 98%, and the content (percentage by weight) of yeast in the dried yeast after pressing is 25%.

[0064] Then use fast paper electrophoresis to track and detect the fermentation products.

[0065] The pH value was adjusted to 2.0 with trichloroacetic acid, the fermentation was terminated, and the ferment...

Embodiment 2

[0105] 1. Fermentation

[0106] First, adopt one-step method to ferment uridine monophosphate (5 liter scale), fermentation conditions: UMP 240g (feeding according to electrophoretic purity 75.3%); Saccharomyces cerevisiae 2.4kg; Glu 312g; NaH 2 PO 4 216g; MgCl 2 57.1g; pH 6.2; H 2 Add O to 5 liters; keep warm at 32°C and ferment statically for 8 hours.

[0107] Brewer's yeast is eliminated yeast in beer production, and the survival rate of yeast cells is more than 90%, and the yeast content (percentage by weight) in the dried yeast after pressing is 20%.

[0108] Then use fast paper electrophoresis to track and detect the fermentation products.

[0109] The pH value was adjusted to 3.0 with trichloroacetic acid to terminate the reaction, and the fermentation conversion rate was 78.9%.

[0110] 2. Microfiltration to remove protein:

[0111] Use an inorganic ceramic membrane with a pore size of 0.1 μm to treat the fermentation broth. Before microfiltration, pass through ...

Embodiment 3

[0148] The basic steps are the same as in Example 1, except that during one-step fermentation, fermentation conditions: UMP 500g (feeding according to electrophoretic purity 75.3%); brewer's yeast (fresh) 3.5kg; Glu 493.8g; NaH 2 PO 4 360g; MgCl 2 5.71g; pH7.0; H 2 Add O to 8 liters; keep warm at 39°C, and ferment statically for 7 hours. The fermentation conversion rate was 77.5%.

[0149] When using 717 chlorine type strong base anion exchange resin for UTP separation and purification, the upper column: pH 2.5, upper column flow rate 26L / hr; low salt elution: 0.02N, pH 2.0NaCl solution; elution flow rate 26L / hr; Salt elution: 0.5N NaCl, pH 2.0 solution; elution flow rate 2.8L / hr.

[0150] Loading volume: 2.36% of the total exchange volume of 717 resin, that is, 40mg UTP / ml resin,

[0151] For acid-thermal degradation, the pH value was first adjusted to 1.2 with 6N hydrochloric acid, heated and boiled for 30 minutes, and detected by high performance liquid phase and pap...

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Abstract

The invention discloses a preparing method of uridine diphosphite, which comprises the following steps: blending uridine monophosphate and beer yeast to ferment; terminating fermenting to obtain the ferment liquid of uridine triphosphate; predisposing ferment liquid; separating and purifying; proceeding acid heat to decompose purified uridine triphosphate; filtering; separating; refining. The invention shortens the predisposing time by two thirds and separating purifying time by one third, which makes receiving rate by over 30%.

Description

technical field [0001] The invention relates to a preparation method of uridine diphosphate, specifically, a method for producing uridine diphosphate by using uridine monophosphate (UMP) as a raw material through a two-step method of fermentation and degradation. Background technique [0002] In the 1960s and 1970s, my country began to use nucleoside monophosphate as raw material and use yeast fermentation to produce nucleoside triphosphate (NTP), mainly producing adenosine triphosphate (ATP) and cytidine triphosphate (CTP). There are many studies on the production of ATP by yeast fermentation. In the 1970s and 1980s, there was a relatively stable fermentation production process, and small-scale production could be carried out. However, there are still many disadvantages in the process of producing ATP, CTP, etc. by fermentation, such as complex process, cumbersome operation, high production cost, and not suitable for large-scale production. The main reasons are as follows:...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/26C12R1/865
Inventor 曾滨饶林凡黎高沃
Owner 北京燕京中科生物技术有限公司
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