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Synthesis method of 2'-deoxyguanosine by adopting nucleoside phosphorylase of brevibacterium acetylium

A technology of nucleoside phosphorylase and Brevibacterium acetyl, applied in the field of synthesizing 2'-deoxyguanosine, can solve the problems of long route and high cost, and achieve the effect of low cost

Inactive Publication Date: 2011-09-07
NANTONG QIUZHIYOU BIOSCI & BIOTECH +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Others also use thymidine and 2,6-diaminopurine to generate 2,6-diamino 2'-deoxynucleoside, and then treat it with adenosine deaminase to obtain 2'-deoxyguanosine, but the route is too long. higher cost

Method used

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  • Synthesis method of 2'-deoxyguanosine by adopting nucleoside phosphorylase of brevibacterium acetylium
  • Synthesis method of 2'-deoxyguanosine by adopting nucleoside phosphorylase of brevibacterium acetylium

Examples

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Effect test

Embodiment 1

[0024] Put 50ml of medium in a 250ml Erlenmeyer flask, which contains 1% beef extract, 1% peptone, 0.5% yeast extract, 0.5% NaCl, pH 7.0, sterilize at 118°C for 30min, and insert Brevibacterium acetyl QD96 after cooling , shake (200r / min) at 30°C for 16h, freeze and centrifuge the culture solution at 4000r / min for 30min, wash with 0.05mol / L phosphate buffer (pH7.0), and centrifuge again, and store the obtained bacteria in cold storage , as a nucleotide phosphorylase.

[0025] In the 100ml round bottom flask, pack 50ml substrate solution, wherein contain guanylic acid and thymidine concentration and be respectively 40mol / L, phosphate buffer is 25mmol / L, thalline addition is 5% (wet weight), in Stir and react in a constant temperature water bath at 60° C. for 2 hours. After the reaction is completed, the cells and a small amount of thymine precipitates are removed by refrigerated centrifugation. Then the solution is detected by HPLC to contain 24mmol / L deoxyguanosine, and its c...

Embodiment 2

[0028] In the 100ml round-bottomed flask, pack 50ml substrate solution, wherein contain guanylic acid and 2'-deoxyuridine concentration and be respectively 40mmol / L, phosphate buffer is 25mmol / L, and thalline addition is 5% (wet weight ), and then operate with embodiment 1. After the reaction, the solution was detected by HPLC, which contained 26 mmol / L deoxyguanosine, and its conversion rate was 65%.

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Abstract

The invention provides a synthesis method of 2'-deoxyguanosine by adopting nucleoside phosphorylase of brevibacterium acetylium, which comprises the following steps of: (1) culturing the brevibacterium acetylium QD96-CGMCCNo.0472; and (2) adding thalli obtained in the step (1) into a substrate solution or reaction, wherein deoxyribose receptors, deoxyribose donors and phosphate buffer solution are contained in the substrate solution, and collecting the 2'-deoxyguanosine from the reaction product. When the enzymes of the brevibacterium acetylium QD96 are used for synthesizing the 2'-deoxyguanosine, the cost can be reduced, and the 2'-deoxyguanosine can be effectively obtained; moreover, the conversion rate can reach more than 60%.

Description

technical field [0001] The present invention relates to a method for synthesizing 2'-deoxyguanosine. Background technique [0002] 2'-Deoxyguanosine is an important raw material for the synthesis of oligodeoxynucleotide and other antiviral and antitumor nucleic acid drugs. Most of the traditional manufacturing methods of nucleoside derivatives use chemical synthesis. The disadvantage of this method is that there are many chemical synthesis steps, high difficulty, long cycle and the generation of isomers. For example, to chemically modify natural nucleosides, it is often necessary to protect the base or ribose active group first, and remove the protecting group after the reaction, so that the overall yield of the synthesis reaction will decrease. Directly condensing bases and ribose by chemical methods will form α-isomers. To obtain the same natural β-compounds, complex separation methods are often required, and the yield is low. Chemical synthesis uses a variety of flammab...

Claims

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Application Information

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IPC IPC(8): C12P19/32C12R1/13
Inventor 邱蔚然曹静李喻周长林邱志云陆长德
Owner NANTONG QIUZHIYOU BIOSCI & BIOTECH
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