Method for decolorizing nucleotide enzymatic hydrolysate

A nucleotide enzymatic solution and pigment technology, applied in the field of separation and purification of 5'-nucleotides, can solve problems affecting the purity and chroma of nucleotide products, reduce the reuse rate of resins, resin poisoning, etc., and achieve Improve appearance color, low price, large adsorption effect

Inactive Publication Date: 2013-10-02
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is a large amount of pigment in the RNA enzymatic hydrolysis solution. If it is directly separated by resin without pretreatment, although the resin will filter out some impurities, a large part of the pigment will remain in the eluent of nucleotides. The post-separation and crystallization procedures of nucleotides bring a lot of pressure, which affects the purity and color of nucleotide products, and the pigments adsorbed on the resin will cause the resin to be poisoned to varying degrees, thereby reducing the reuse rate of the resin

Method used

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  • Method for decolorizing nucleotide enzymatic hydrolysate
  • Method for decolorizing nucleotide enzymatic hydrolysate
  • Method for decolorizing nucleotide enzymatic hydrolysate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1: Preparation of nucleotide enzymatic hydrolysis solution.

[0048] The RNA enzymatic hydrolysis process uses nuclease p1 to act on the RNA solution to obtain 4 kinds of 5'-mononucleotides, namely 5'-adenylic acid (AMP), 5'-cytidylic acid (CMP), 5'- Guanylate (GMP) and 5'-Uridylic acid (UMP). Specific steps are as follows:

[0049] (1) Prepare 600ml of 6% RNA solution with pH=5.5: first use 1mol / L NaOH solution to adjust the pH to about 13, then add RNA powder (36g) to dissolve, then use 1mol / L NaOH solution to adjust the pH to 5.5, place in a 70°C water bath and preheat to 70°C.

[0050] (2) Add 50ml of enzyme solution preheated at 70°C for half an hour to the RNA solution, shake well, and enzymatically digest at 70°C for 3h50min.

[0051] (3) Add 0.2% (1.2g) activated carbon and continue enzymatic hydrolysis for 50 minutes.

[0052] (4) Cool to 45° C., centrifuge at 4000 rpm for 10 minutes, and filter with suction to obtain RNA enzymatic hydrolysis sol...

Embodiment 2

[0053] Embodiment 2: the acquisition of ultra-high cross-linked resin SX-01.

[0054] In a clean and dry 2000mL three-neck flask, add 200g of dry chloromethylated polystyrene resin (chlorine balls), add 1200g of o-nitroethylbenzene, stir and mix well, fully swell at room temperature for more than 2 hours, stir and add Methylamine 4 times the weight of chlorine balls, then add 20g of anhydrous ferric chloride as a catalyst, program temperature rise to 135-150°C under stirring, and then carry out Friedel-Crafts post-crosslinking reaction at this temperature for a certain period of time to control the residual chlorine content At 1-3%, stop the reaction, suck out the mother liquor after cooling, and repeatedly extract the resin with industrial alcohol until the extraction effluent is colorless and clear, then wash the resin with 4% hydrochloric acid and distilled water until it is neutral, and dry it for later use.

[0055] The obtained product is in the shape of brown round part...

Embodiment 3

[0056] Embodiment 3: the pretreatment of ultrahigh cross-linking resin SX-01.

[0057] Rinse the resin with 2BV of ethanol at a flow rate of 2BV / h→wash away the ethanol with deionized water→2BV0.1mol / L HCl solution wash the resin at a flow rate of 1BV / h→wash with water until neutral→use 2BV0.1mol / L Rinse the resin with NaOH solution at a flow rate of 1BV / h→wash with water to neutral→wash the resin with 2BV0.1mol / L HCl solution at a flow rate of 1BV / h→wash with water to neutral.

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Abstract

The invention relates to a method for decolorizing nucleotide enzymatic hydrolysate. The method comprises the following steps of: performing pigment adsorption on the nucleotide enzymatic hydrolysate through hyper-crosslinked resin SX-01; and after adsorption is saturated, and regenerating the hyper-crosslinked resin SX-01 by using a regenerant, wherein the hyper-crosslinked resin SX-01 has the following structure units: the skeleton of the resin is polystyrene, the average grain size is 0.3 to 1 mm, the water content is 32 to 49 percent, the aperture is 1.5 to 4 nm, the most probable aperture is 1.8 nm, the average specific surface area is 950 to 1,800 m<2> / g, and the average pore volume is 0.54 to 0.82 cm<3> / g. By adoption of the method, the decolorization ratio of the enzymatic hydrolysate reaches over 80 percent, the treatment quantity reaches 11 times of bed volume (BV), the loss ratio of four kinds of nucleotides is lower than 5 percent, and the consumption quantity of the regenerant is 5 BV.

Description

technical field [0001] The invention belongs to the field of separation and purification of 5'-nucleotides, and in particular relates to the application of super-high cross-linked resin SX-01 in the decolorization of nucleotide enzymatic hydrolysis solution. Background technique [0002] Nucleosides and nucleotides are the basis of biological genetic information DNA and RNA, and play an important role in the material metabolism and energy conversion of biological cells. 5'-mononucleotides and their derivatives have very important uses in the fields of genetic engineering, medicine, food, agricultural production and scientific research. Adding nucleotides in health care products and infant food is beneficial to the human body to supplement nucleotides, making nucleotide metabolism in a vigorous state, which can achieve the purpose of resisting aging, prolonging human life, improving human immune function and treating diseases; in agriculture On the one hand, nucleotides have...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/00C07H1/06C07H19/10C07H19/20
Inventor 应汉杰焦朋飞吴菁岚刘俊周精卫张宇
Owner NANJING TECH UNIV
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