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Method for preparing nucleotide using phosphoric acid circulation

A manufacturing method and nucleotide technology, applied in the field of nucleotide manufacturing, can solve problems such as difficulty in obtaining high phosphorylation yield and the like

Active Publication Date: 2008-09-17
AJINOMOTO CO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At this time, if the pH is only adjusted before the reaction, the pH is biased toward the alkaline side, and there is a problem that it is difficult to obtain a high phosphorylation yield.

Method used

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  • Method for preparing nucleotide using phosphoric acid circulation
  • Method for preparing nucleotide using phosphoric acid circulation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Using basic sodium condensed phosphate with an average degree of polymerization of about 3 to 4 to increase the phosphorylation yield when adding phosphoric acid to nucleosides

[0040] (1) Preparation of bacteria containing acid phosphatase

[0041] Escherichia coli JM 109 / pEPI370 carrying the plasmid pEPI370 shown in Reference Example 1 described in JP-A-2002-289 was inoculated in 50 ml of L medium, cultured at 37° C. for 16 hours, and the culture solution was used as Enzyme solution is used. The enzyme solution contains bacterial cells containing acid phosphatase. Enzyme solutions in the following examples all use enzyme solutions prepared by this method.

[0042] (2) Preparation of phosphoric acid addition reaction raw material slurry

[0043] The method disclosed in JP 2002-289 A was used. That is, the guanosine crystal is made into a 27.1wt% slurry in water, and the pulverization process of the crystal is carried out by a pulverizer (DYNO MILL produ...

Embodiment 2

[0047] Embodiment 2: reclaim the mixture of orthophosphate and pyrophosphate from the manufacturing process of 5'-ribonucleotide disodium, and regenerate the recovered phosphate into condensed sodium phosphate with an average degree of polymerization of about 3 to 4, Re-use in the production of 5'-ribonucleotide disodium

[0048] (1) Response to nucleoside addition of phosphate

[0049] Using the guanosine slurry after the same crushing treatment as in Example 1, 92g inosine, 79g guanosine and 228g sodium tripolyphosphate were suspended in 760g water, and then 95g35% hydrochloric acid was added to adjust the pH to 4.5. Add 228ml of enzyme solution at 0.5°C to start the reaction. During the reaction, 30 g of 35% hydrochloric acid was intermittently added to maintain the pH at 4.5. About 19 hours after the start of the reaction, an enzyme reaction solution containing 289 g of 5'-ribonucleotides was obtained.

[0050] (2) Phosphate crystallization removal process

[0051] The...

Embodiment 3

[0056] Embodiment 3: Reclaim the mixture of orthophosphate and pyrophosphate from the manufacturing process of 5'-disodium inosinate, condense and regenerate the recovered phosphate into condensed sodium phosphate with an average degree of polymerization of about 3 to 4, and again When used in the production of disodium 5'-inosinate

[0057] (1) Response to nucleoside addition of phosphate

[0058] Suspend 55g of inosine and 54g of sodium tripolyphosphate in 97g of water, add 15g of 35% hydrochloric acid, adjust the pH to 4.5, add 50ml of enzyme solution at 35±0.5°C, and start the reaction. During the reaction, 12 g of 35% hydrochloric acid was intermittently added to maintain the pH at 4.5. About 26 hours after the start of the reaction, an enzyme reaction liquid containing 101 g of disodium 5'-inosinate was obtained.

[0059] (2) Phosphate crystallization removal process

[0060] The pH of the above reaction solution was adjusted to 9.7 with 34% sodium hydroxide aqueous s...

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PUM

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Abstract

The invention aims to provide a manufacturing method of nucleotide by phosphorus cycle. In a manufacturing method that adds phosphoric acid molecules of condensed phosphoric acid to nucleotide mixture or inosine by acidic phosphatase in particular manufacturing of 5'-disodium ribonucleotide and 5'-disodium inosinate, orthophosphoric acid after reaction is recycled and condensed again as raw materials of addition reaction of phosphoric acid, which improves phosphorylation yield (nucleic acid yield) and utilization rate of phosphoric acid. The manufacturing method of nucleotide is characterized in that when the nucleotide is phosphorylated by polyphosphoric acid and acidic phosphatase, the condensed phosphate is used as polyphosphoric acid, the reaction of phosphorylation processed at a certain pH controlled continuously in a scale of 4.10 to 5.15.

Description

technical field [0001] The present invention relates to a method for producing nucleotides, such as 5'-ribonucleotide disodium or 5'-inosinate disodium, which uses acid phosphatase to make condensed phosphate act on nucleosides such as guanosine and inosine and Manufactured by adding phosphoric acid to it. Background technique [0002] Acid phosphatase (JP-A-7-231793, JP-A-9-37785, JP-A 2001-245676, JP-A 2001-136984) can cleave one of the condensed phosphoric acids and add it to the nucleus Glycosides. Orthophosphoric acid, on the other hand, cannot be used for such purposes. Therefore, when using condensed phosphoric acid such as acid pyrophosphoric acid with a degree of condensation of 2, after the reaction is over, a large amount of orthophosphoric acid produced by the reaction remains in the liquid at the end of the reaction, and the orthophosphoric acid in the waste liquid after separating and purifying nucleotides is Massive, unmanageable problems. [0003] On the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/30
Inventor 永井秀忠星田和纪小池功东森郁彦
Owner AJINOMOTO CO INC
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