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Preparation method of uridine diphosphate glucose

A technology of uridine diphosphate glucose and uridine diphosphate, which is applied in the field of preparation of uridine diphosphate glucose, and can solve problems such as difficult separation and aggravated separation process burden

Active Publication Date: 2015-04-29
SHANGHAI ZHAOWEI TECH DEV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of using the yeast system to synthesize UTP and UDPG, a certain amount of UDP-Gal will be produced, and it is difficult to separate UDPG and UDP-Gal, which increases the burden on the subsequent separation process

Method used

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  • Preparation method of uridine diphosphate glucose
  • Preparation method of uridine diphosphate glucose
  • Preparation method of uridine diphosphate glucose

Examples

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preparation example Construction

[0055] The method for preparing uridine diphosphate glucose (UDPG) comprises: in an in vitro (extracellular) reaction system, using uridine triphosphate (UTP) or its salt (such as sodium salt) and maltodextrin as substrates , add inorganic ions, dTT and Tris, and use uridine diphosphate glucose pyrophosphorylase and maltodextrin phosphorylase recombinantly expressed in Escherichia coli as catalysts to produce uridine diphosphate glucose through biotransformation.

[0056] In the preparation process of UDPG, the UDPG pyrophosphorylase and maltodextrin phosphorylase added in the reaction system can be derived from animals, plants, microorganisms, etc., and the source is not particularly limited, and enzymes from any source can be used . With the development of genetic engineering technology, the use of Escherichia coli to recombine and express the target protein is simpler and feasible than animal and plant extraction. Escherichia coli cells expressing UDPG pyrophosphorylase an...

Embodiment 1

[0077] Embodiment 1, press yeast to prepare UDPG

[0078] (1) Pretreatment of waste beer yeast

[0079] Take 20 kg of fresh waste beer yeast from the brewery, press filter with plate and frame, blow dry, and collect 2.5 kg of filter cake.

[0080] (2) Pressed yeast permeability treatment

[0081] According to the amount of 5ml of xylene added per kilogram of filter cake, 12.5ml of xylene was added to 2.5kg of yeast filter cake, dispersed and mixed at a high speed, and stored at -20°C.

Embodiment 2

[0082] The preparation of embodiment 2, UTP

[0083] UMP·Na is prepared in a reactor with a capacity of 20L 2 300 grams, 10L of reaction solution consisting of 150 grams of glucose, 150 grams of potassium dihydrogen phosphate, 25 grams of magnesium sulfate, and 1.8 kg of pressed yeast, adjust the pH to 7.0 with sodium hydroxide, and react at 37°C for 8 hours, then add hydrochloric acid to adjust PH=2.5, after resting for 30 minutes, high-speed refrigerated centrifugation (12000rpm, 30min), got UTP Na 3 358 grams.

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Abstract

The invention relates to a preparation method of uridine diphosphate glucose (UDPG), which comprises the steps of simply, conveniently, economically, reasonably and greatly synthesizing the UDPG by the combined action of yeast and genetically engineered bacteria, performing purification via resin separation, alcohol precipitation and the like, and obtaining a target compound with very high purity.

Description

technical field [0001] The invention belongs to the field of chemical and biological engineering; more specifically, the invention relates to a preparation method of uridine diphosphate glucose (UDPG). Background technique [0002] Sugar nucleotides play a very good role in the synthesis of functional oligosaccharides and structural modification of proteins, antibiotics, flavonoids, and artemisinin. [0003] Uridine diphosphate glucose (UDPG) is the earliest studied sugar nucleotide, which can be used as a precursor for the synthesis of other sugar nucleotides, for example, UDP-Gal can be generated under the action of UDPG-4′-epimerase, UDP-GlcA is generated under the action of UDPG dehydrogenase, etc.; the modification of flavonoids is mainly carried out with UDPG as the substrate, under the action of glycosyltransferase, such as luteolin, quercetin, naringenin, Catechin, etc. [Chinese Natural Medicine, 2010, 8(5), 389-400]. [0004] The synthesis of UDPG is mainly chemic...

Claims

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Application Information

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IPC IPC(8): C12P19/30C12N9/12C12N9/10C12N15/70C12R1/19
Inventor 李新亮姚峰
Owner SHANGHAI ZHAOWEI TECH DEV
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