Unlock instant, AI-driven research and patent intelligence for your innovation.

Process for producing pentose-5-phosphate ester

A technology of phosphate ester and acid phosphatase, which is applied in fermentation and other directions, and can solve the problems of not using acid phosphatase

Active Publication Date: 2006-12-20
MITSUI CHEM INC
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far there are no examples of pentose sugar phosphorylation using acid phosphatase

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Process for producing pentose-5-phosphate ester
  • Process for producing pentose-5-phosphate ester

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0039] In the preparation method of the present invention, cells derived from microorganisms and higher organisms with the ability to generate acid phosphatase, the cells themselves that have undergone shape and quality transformation using the gene encoding acid phosphatase, and the broken product of the above cells can also be used. The cells, the cell fragments, and the fractions containing the acid phosphatase activity purified by ammonium sulfate precipitation or column chromatography are used as immobilized products formed on the carrier.

[0040] The molar ratio of the pentose sugar and the phosphoric acid donor used in the above reaction is not particularly limited, and usually, the reaction is carried out under the condition that one of them is more than 1 times the molar ratio of the other. For example, the reaction is carried out under the conditions that the phosphoric acid donor is more than 1 times the mole and less than or equal to 20 times the mole of the pentos...

reference example 1

[0059] Based on the known acid phosphatase sequence derived from Shigella flexneri 2a YSH6000, the two primers shown in SEQ ID NO: 1 and SEQ ID NO: 2 were made to contain the acidic acid phosphatase encoding derived from Shigella flexneri 2a YSH6000 The phosphatase gene plasmid was used as a template for PCR. Prepare a reaction consisting of 10mM KOD-plus buffer, 1.5μM forward and reverse primers, 1mM magnesium sulfate, 0.2mM dNTPs, 2U KOD-plus polymerase (TOYOBO), 50ng / μL template DNA solution. After holding at 94° C. for 2 minutes, a thermal cycle of holding at 94° C. for 30 seconds, 60° C. for 30 seconds, and 68° C. for 1 minute was performed for 30 cycles, and finally, 68° C. was held for 10 minutes. As a result, an amplified fragment of about 0.75 kb was obtained. The resulting fragment was treated with PstI / BamHI and ligated with pUC19. Transform the E. coli DH5α strain with the prepared plasmid, so as to make an expression strain of phosphatase activity.

[0060] Pr...

Embodiment 1

[0062] In the solution containing 100mM acetate buffer (pH3.5), 100mM pyrophosphate and potassium pyrophosphate mixed solution (pH3.5), 100mM D-2-deoxyribose solution, add to make the reaction solution The activity value reached 7.3U / mL (5 mg wet bacteria / mL) and the bacteria liquid prepared by the cultured bacteria obtained in Reference Example 1 was reacted at 37°C for 1 hour, and the reaction liquid was analyzed by HPLC. 1.5 mM D-2-deoxyribose-5-phosphate.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

A pentose such as ribose, arabinose, 2-deoxyribose, 1-methoxy-2-deoxyribose or the like is reacted with a phosphoric acid donor such as a pyrophosphoric acid or an alkali metal salt thereof in the presence of an acid phosphatase. Thus, a pentose-5-phosphate ester can be produced, such as a ribose-5-phosphate ester, an arabinose-5-phosphate ester, a 2-deoxyribose-5-phosphate ester, a 1-methoxy-2-deoxyribose-5-phosphate ester or the like.

Description

technical field [0001] The present invention relates to the preparation method of pentose-5-phosphate. Specifically, it relates to a method for producing pentose-5-phosphate used as a starting material for synthesizing nucleosides as one of raw materials for the production of pharmaceuticals and functional chemicals. Background technique [0002] Pentose-5-phosphate is useful as a starting material for the synthesis of nucleosides. For example, it is disclosed to use phosphopentose mutase (phosphopentomutase) to change 2-deoxyribose-5-phosphate into 2-deoxyribose-1-phosphate, and then utilize nucleoside phosphorylase to make it react with various nucleic acid bases A method for preparing various deoxyribonucleosides by glycosylation of a group (Patent Document 1). The 2-deoxyribose-5-phosphate used in this production method is prepared by enzymatic hydrolysis of DNA, but this preparation method uses DNA as a raw material, which is expensive and requires many separation and...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/44
CPCC12P19/44C12P7/62C12P7/64C12P19/00
Inventor 甲斐庆一郎三宅仁基及川利洋
Owner MITSUI CHEM INC