1. A process for producing high-purity ribose-1-phosphate,

the method comprises the following steps: adding nucleoside and nucleoside phosphorylase into purified water, regulating pH and temperature of the reaction system, stirring for reaction, passing the reaction solution through strong acidic cation resin, and collecting the effluent.

2. The process for producing high-purity ribose 1-phosphate according to claim 1,

the adding amount mass ratio of the nucleoside to the nucleoside phosphorylase is (3-7.5): 1, preferably 5: 1.

3. the process for producing high-purity ribose 1-phosphate according to claim 1,

the pH of the reaction system is controlled to 3 to 10, preferably 4.5 to 9.

4. The process for producing high-purity ribose 1-phosphate according to claim 1,

the temperature of the reaction system is adjusted to 0-100 ℃, preferably 32-58 ℃.

5. The process for producing high-purity ribose 1-phosphate according to any of claims 1 to 4,

the nucleoside is one of cytidine, adenosine, guanosine, uridine, inosine, xanthosine and thymidine.

6. The process for producing high-purity ribose 1-phosphate according to claim 5,

the nucleoside phosphorylase is one of cytidine phosphorylase, adenosine phosphorylase, guanosine phosphorylase, uridine phosphorylase, inosine phosphorylase, xanthosine phosphorylase and thymidine phosphorylase.

7. The process for producing high-purity ribose 1-phosphate according to any of claims 1 to 6,

the addition mass of the purified water is 6 to 15 times, preferably 7.5 to 9 times that of the nucleoside.

8. The process for producing high-purity ribose 1-phosphate according to any of claims 1 to 7,

and detecting the reaction conversion rate in real time during the stirring reaction process, stopping the reaction when the reaction conversion rate reaches 55-65%, and allowing the reaction solution to pass through strong-acid cation resin.

9. A ribose-1-phosphate of high purity obtained by the method of any one of claims 1 to 8.

Background

Ribose-1-phosphate is an important substance, can be effectively used as a raw material for synthesizing nucleoside, and has great application value in the field of medicine.

In the prior art, the preparation method of ribose-1-phosphate mainly comprises the following two methods:

a method for preparing various deoxyribonucleosides by converting 2-deoxyribose-1-phosphate into 2-deoxyribose-1-phosphate by using pentosemutase, and then glycosylating the 2-deoxyribose-1-phosphate with various nucleic acid bases by using nucleoside phosphorylase; the 2-deoxyribose-1-phosphate used in this production method is prepared by enzymatic hydrolysis of DNA, but this preparation method uses DNA as a raw material and is expensive, and the steps of separation and purification are complicated.

Another production method is a method in which deoxyribokinase is allowed to act on 2-deoxyribose and Adenosine Triphosphate (ATP) as a phosphate donor to produce-2-deoxyribose-5-phosphate; ATP used in this method is expensive, and if polyphosphoric acid, which is cheaper than ATP, is used as a phosphate donor, the method of phosphorylating an organic hydroxy compound with alkaline phosphatase derived from calf intestine cannot clearly identify which of the hydroxyl groups present in the saccharide molecule is introduced with a phosphate group.

In view of this, how to further optimize the preparation method of ribose-1-phosphate based on the prior art, improve the overall yield and purity of ribose-1-phosphate product, and reduce the cost is a key technical problem to be solved urgently by the technical personnel in the field.

Disclosure of Invention

Aiming at the technical problems in the prior art, the invention provides a preparation method of high-purity ribose-1-phosphate.

The invention adopts the following technical scheme:

a method for preparing high-purity ribose-1-phosphate, comprising:

adding nucleoside and nucleoside phosphorylase into purified water, regulating pH and temperature of the reaction system, stirring for reaction, passing the reaction solution through strong acidic cation resin, and collecting the effluent.

The specific process route is as follows:

in the above-mentioned process for producing high-purity ribose-1-phosphate, the amount of the nucleoside to the nucleoside phosphorylase added is (3 to 7.5): 1, preferably 5: 1.

in the above-mentioned process for producing high-purity ribose-1-phosphate, the pH of the reaction system is controlled to 3 to 10, preferably 4.5 to 9.

In the above-mentioned process for producing high-purity ribose-1-phosphate, the temperature of the reaction system is adjusted to 0 to 100 ℃ and preferably 32 to 58 ℃.

Further, in the above method for producing high-purity ribose 1-phosphate, the nucleoside is one of cytidine, adenosine, guanosine, uridine, inosine, xanthosine, and thymidine.

Still further, in the above method for producing high-purity ribose 1-phosphate, the nucleoside phosphorylase is one of cytidine phosphorylase, adenosine phosphorylase, guanosine phosphorylase, uridine phosphorylase, inosine phosphorylase, xanthosine phosphorylase, and thymidine phosphorylase.

Still further, in the above-mentioned process for producing high-purity ribose-1-phosphate, the added mass of the purified water is 6 to 15 times, preferably 7.5 to 9 times the added mass of the nucleoside.

Still further, the above-mentioned process for producing high-purity ribose-1-phosphate comprises the steps of detecting the reaction conversion rate in real time during the course of the stirring reaction, stopping the reaction when the reaction conversion rate reaches 55 to 65%, and passing the reaction solution through a strongly acidic cationic resin.

The invention also provides the high-purity ribose-1-phosphate prepared by the preparation method.

Compared with the prior art, the invention has the beneficial effects that:

the invention adopts cheap and easily available nucleoside as raw material, realizes the preparation of ribose-1-phosphate by one-step enzyme method under the catalysis of nucleoside phosphorylase, and has low cost and high yield; in addition, the reaction solution obtained by the reaction is passed through strong acid cation resin, so that the high-purity ribose-1-phosphate can be obtained, and the practical application prospect is good.

Detailed Description

The present invention is further described in detail below with reference to specific examples so that those skilled in the art can more clearly understand the present invention.

The following examples are provided only for illustrating the present invention and are not intended to limit the scope of the present invention.

All other embodiments obtained by a person skilled in the art based on the specific embodiments of the present invention without any inventive step are within the scope of the present invention.

In the embodiments of the present invention, all components are commercially available products well known to those skilled in the art unless otherwise specified.

In the examples of the present invention, unless otherwise specified, all technical means used are conventional means well known to those skilled in the art.

Example 1

The embodiment of the invention provides a preparation method of high-purity ribose-1-phosphate.

1000g of cytidine, 559g of potassium dihydrogen phosphate, 200g of cytidine phosphorylase and 8000g of purified water are added into a 10L glass reaction kettle, stirring is started, dissolution is carried out, the temperature is controlled at 37 ℃, the reaction conversion rate is detected by HPLC to reach 60%, and the reaction is stopped.

The reaction solution was passed through 20L of cationic resin, and the effluent was collected, i.e., an aqueous solution of ribose-1-phosphate having a purity of 95.8 wt%.

Example 2

The embodiment of the invention provides a preparation method of high-purity ribose-1-phosphate.

Adding 1000g of adenosine, 509g of monopotassium phosphate, 200g of adenosine phosphorylase and 8000g of purified water into a 10L glass reaction kettle, starting stirring and dissolving, controlling the temperature to 37 ℃, detecting the reaction conversion rate to 60% by HPLC, and stopping the reaction.

The reaction solution was passed through 20L of cationic resin, and the effluent was collected, i.e., an aqueous solution of ribose-1-phosphate having a purity of 96.2 wt%.

Example 3

The embodiment of the invention provides a preparation method of high-purity ribose-1-phosphate.

1000g of guanosine, 480g of monopotassium phosphate, 200g of guanosine phosphorylase and 8000g of purified water are added into a 10L glass reaction kettle, stirring is started, dissolution is carried out, the temperature is controlled at 37 ℃, the reaction conversion rate is detected by HPLC to reach 60%, and the reaction is stopped.

The reaction solution was passed through 20L of cationic resin, and the effluent was collected, i.e., an aqueous solution of ribose-1-phosphate having a purity of 97.1 wt%.

Example 4

The embodiment of the invention provides a preparation method of high-purity ribose-1-phosphate.

1000g of uridine, 557g of monopotassium phosphate, 200g of uridine phosphorylase and 8000g of purified water are added into a 10L glass reaction kettle, stirring is started, dissolution is carried out, the temperature is controlled at 37 ℃, the reaction conversion rate is detected to be 60% by HPLC, and the reaction is stopped.

The reaction solution was passed through 20L of cationic resin, and the effluent was collected, i.e., an aqueous solution of ribose-1-phosphate having a purity of 95.6 wt%.

Example 5

The embodiment of the invention provides a preparation method of high-purity ribose-1-phosphate.

1000g of inosine, 507g of monopotassium phosphate, 200g of inosine phosphorylase and 8000g of purified water are added into a 10L glass reaction kettle, stirring is started, dissolution is carried out, the temperature is controlled at 37 ℃, the reaction conversion rate is detected to be 60% by HPLC, and the reaction is stopped.

The reaction solution was passed through 20L of cationic resin, and the effluent was collected, i.e., an aqueous solution of ribose-1-phosphate having a purity of 97.3 wt%.

Example 6

The embodiment of the invention provides a preparation method of high-purity ribose-1-phosphate.

1000g of xanthosine, 478g of potassium dihydrogen phosphate, 200g of xanthosine phosphorylase and 8000g of purified water are added into a 10L glass reaction kettle, stirring is started, dissolution is carried out, the temperature is controlled at 37 ℃, the reaction conversion rate is detected by HPLC to reach 60%, and the reaction is stopped.

The reaction solution was passed through 20L of cationic resin, and the effluent was collected, i.e., an aqueous solution of ribose-1-phosphate having a purity of 96.4 wt%.

Example 7

The embodiment of the invention provides a preparation method of high-purity ribose-1-phosphate.

1000g of thymidine, 561g of potassium dihydrogen phosphate, 200g of thymidine phosphorylase and 8000g of purified water are added into a 10L glass reaction kettle, stirring is started, dissolution is carried out, the temperature is controlled at 37 ℃, the reaction conversion rate is detected by HPLC to reach 60%, and the reaction is stopped.

The reaction solution was passed through 20L of cationic resin, and the effluent was collected, i.e., an aqueous solution of ribose-1-phosphate having a purity of 95.7 wt%.

It should be noted that the above examples are only for further illustration and description of the technical solution of the present invention, and are not intended to further limit the technical solution of the present invention, and the method of the present invention is only a preferred embodiment, and is not intended to limit the protection scope of the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.