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A kind of production method of nucleoside drug intermediate 2'-deoxyguanosine

A technology of deoxyguanosine and its production method, applied in the biological field, can solve problems such as low solubility and increase the solubility of guanine, and achieve the effect of increasing solubility and improving conversion rate

Active Publication Date: 2018-07-27
乐山市佰尔特生物工程合伙企业(有限合伙)
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method applies the two-phase aqueous system to the process of biocatalytic preparation of 2'-deoxyguanosine, which solves the bottleneck problem of low solubility of guanine in the conversion system in the past, can greatly increase the solubility of guanine, and greatly improve the conversion rate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Biocatalytic synthesis of 2'-deoxyguanosine

[0030] 1) Collection and pretreatment of enzyme-producing bacteria

[0031] Lactobacillus fermentum (Lactobacillus fermentum)CGMCC 1.1880 was propagated and grown under the above-mentioned known enzyme-producing culture conditions and enriched in deoxynucleoside ribosyltransferase, and 1 liter of enzyme-producing fermentation broth was obtained, which was collected by centrifugation (centrifugation conditions: 6000 rpm, 20 minutes) to obtain 57 grams of thalline slurry, then thalline was remade into bacterial suspension with 500 milliliters of deionized water and stirred for 10 minutes to wash the thalline, then centrifuged to collect the thalline under the same centrifugal conditions as above to obtain 53 grams of washed thalline bacterial pulp. Then wash with 1 liter of 0.5M phosphate buffer (pH 8.0), and then centrifuge under the same conditions to obtain 50 g of thalline slurry. The bacterium slurry is the ...

Embodiment 2

[0036] Embodiment 2: Biocatalytic method produces 2'-deoxyguanosine

[0037] PEG10000 in Example 1 was replaced with PEG8000, and other conditions were the same as in Example 1. After the reaction, HPLC analysis showed that the conversion rate of β-thymidine was 60%, and the yield of product 2'-deoxyguanosine was 10.8 g / L.

Embodiment 3

[0038] Embodiment 3: Biocatalytic method produces 2'-deoxyguanosine

[0039] PEG10000 in Example 1 was replaced with PEG4000, and other conditions were the same as in Example 1. After the reaction, HPLC analysis showed that the conversion rate of β-thymidine was 46%, and the yield of product 2'-deoxyguanosine was 8.2%. g / L.

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Abstract

The invention belongs to the field of biotechnology, relates to the technical field of preparing 2'-deoxyguanosine by a biocatalysis method, and specifically relates to a production method of a nucleoside drug intermediate 2'-deoxyguanosine. The method uses β-thymidine Using guanine as a raw material, using Lactobacillus fermentum cell slurry as a biocatalyst, the biocatalytic reaction is carried out under the polyethylene glycol 10000 / dipotassium hydrogen phosphate two-phase system to obtain the 2'-deoxyguanosine product. Under the optimal conditions, the amount of substrate β-thymidine added reached 16.5g / L, the conversion rate of thymidine reached 89%, and the product 2'-deoxyguanosine reached 16.1g / L.

Description

technical field [0001] The invention belongs to the field of biotechnology, relates to the technical field of preparing 2'-deoxyguanosine by a biocatalysis method, and specifically relates to a production method of a nucleoside drug intermediate 2'-deoxyguanosine. Background technique [0002] 2'-Deoxyguanosine is a natural deoxynucleoside, which can be directly used to prepare combined deoxynucleoside drugs or as a chemical reagent for biochemical research; at the same time, it can be used as an intermediate for the synthesis of some antiviral nucleoside drugs and Molecular markers, such as literature (Journal of Medicinal Chemistry, 1985, 28: 1194-1198) Synthesis of 8-bromo-2'-deoxyguanosine and 8-hydroxy-2'-deoxyguanosine from 2'-deoxyguanosine raw materials and other important molecular markers; literature (Tetrahedron Letters, 1982, 23 (22): 2253-2256, 1982 Tetrahedron Letters, 1990, 31 (3): 319-322) reported the 6-position derivatization of 2'-deoxyguanosine substance...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/40C12R1/225
Inventor 李竹谌京梅
Owner 乐山市佰尔特生物工程合伙企业(有限合伙)
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