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Enzymatic synthesis method for L-selenium-methyl selenocysteine

A technology of methylselenocysteine ​​and selenocysteine, applied in biochemical equipment and methods, enzymes, transferases, etc., can solve the problem of difficulty in large-scale actual production, complicated steps and harsh reaction conditions. and other problems, to achieve the effect of high practical promotion value, specificity and efficiency, and simple steps

Inactive Publication Date: 2018-11-23
由永峰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] These artificial synthesis methods have their own shortcomings, such as: complicated steps, or lengthy process, or harsh reaction conditions, or high raw material prices, or involve highly toxic raw materials, etc.
Therefore, most of them are difficult to put into large-scale practical production
Therefore, L-selenium-methylselenocysteine ​​on the market is still very expensive, up to 300,000 yuan per kilogram

Method used

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  • Enzymatic synthesis method for L-selenium-methyl selenocysteine
  • Enzymatic synthesis method for L-selenium-methyl selenocysteine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 (continuous process)

[0035] Step 1: 1.0 g of selenocysteine-selenomethyltransferase was immobilized on the surface of 100 g of polymer microparticles in advance, and packed into a 200 mL glass column.

[0036] Step 2: Take 200mL of water and pass nitrogen gas at room temperature for 30min to remove oxygen. Add L-selenocystine (3.34g, 10mmol) and β-mercaptoethanol (14mL, 200mmol), and stir with a magnet for 30min until L-selenocystine is completely dissolved and reduced to L-selenocysteine . During the process, the liquid level was kept under nitrogen.

[0037] Step 3: Take 0.8L ammonium acetate buffer solution (50mM, pH 6.0), add S-methyl-L-methionine (3.60g, 22mmol), magnesium acetate (0.3g, 2.1mmol), stir to dissolve, and then add step 2 The resulting L-selenocysteine ​​solution.

[0038]Step 4: Keep the above solution (step 3) at 37°C and keep the liquid level under nitrogen, and pump it into the glass column (step 1) with the immobilized enzyme wi...

Embodiment 2

[0040] Embodiment 2 (continuous process)

[0041] Step 1: 0.5 g of selenocysteine-selenomethyltransferase was immobilized on the surface of 100 g of polyester fiber in advance, and packed into a 200 mL glass column.

[0042] Step 2: Take 200mL of water and pass nitrogen gas at room temperature for 30min to remove oxygen. Add L-selenocystine (1.67g, 5mmol) and sodium borohydride (3.78g, 100mmol), and stir with a magnet for 30min until L-selenocystine is completely dissolved and reduced to L-selenocysteine . During the process, the liquid level was kept under nitrogen.

[0043] Step 3: Take 0.8L ammonium acetate buffer solution (20mM, pH 7.0), add S-methyl-L-methionine (1.80g, 11mmol), magnesium acetate (0.3g, 2.1mmol), stir to dissolve, and then add step 2 The resulting L-selenocysteine ​​solution.

[0044] Step 4: Keep the above solution (step 3) at 30° C. with the liquid level under nitrogen, and pump it into the glass column (step 1) with the immobilized enzyme with a sm...

Embodiment 3

[0046] Embodiment 3 (batch process)

[0047] Step 1: Immobilize 0.25 g of selenocysteine-selenomethyltransferase on the surface of 50 g of polymer microparticles in advance, and set aside.

[0048] Step 2: Take 200mL of water and pass nitrogen gas at room temperature for 30min to remove oxygen. Add L-selenocystine (1.67g, 5mmol) and β-mercaptoethanol (7.0mL, 100mmol), and stir with a magnet for 30min until L-selenocystine is completely dissolved and reduced to L-selenocysteine acid. During the process, the liquid level was kept under nitrogen.

[0049] Step 3: Take 0.8L of ammonium acetate buffer solution (25mM, pH 6.0), add S-methyl-L-methionine (1.80g, 11mmol), magnesium acetate (0.3g, 2.1mmol), and stir with a magnet to dissolve.

[0050] Step 4: Mix immobilized enzyme (step 1), L-selenocysteine ​​solution (step 2) and S-methyl-L-methionine buffer solution (step 3). The liquid level was maintained under nitrogen at 37°C with slow agitation using a magnet. After 16 hour...

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Abstract

The invention provides an enzymatic synthesis method for L-selenium-methyl selenocysteine. In particular, L-selenocysteine and S-methyl-L-methionine are taken as substrates, and immobilized selenocysteine-selenium methyltransferase (EC 2.1.1.280) is taken as a catalyst to synthesize L-selenium-methyl selenocysteine in a continuous or batch process. The product can be taken as an organic selenium food additive or a supplement additive after purification. A synthesis reaction is performed under a mild condition in an environment-friendly way, and has the characteristics of specificity, high efficiency, simple steps, easiness in control, no production of harmful impurities in the reaction and low substrate cost. The enzymatic synthesis method is suitable for producing the L-selenium-methyl selenocysteine in an environment-friendly and low-cost manner.

Description

technical field [0001] The invention relates to the field of amino acid production, in particular to an enzymatic synthesis method of L-selenium-methylselenocysteine. Background technique [0002] Selenium is an essential trace element for the human body, and it is a constituent element of selenocysteine. Selenocysteine, known as the twenty-first amino acid, is used in the synthesis of more than twenty important proteins in the human body. Selenium is scarce in nature, and its geographical distribution is uneven. Selenium deficiency can lead to a variety of diseases, the more common ones are Keshan disease, Kaschin-Beck disease, heart disease, thyroid disease and low immune function. [0003] Selenium deficiency in the human body can be supplemented by selenium nutritional fortifiers. The newest selenium booster is L-selenium-methylselenocysteine. It has a clear chemical structure, is easily absorbed by the human body, has no obvious accumulation toxicity, and has good q...

Claims

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Application Information

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IPC IPC(8): C12P13/12C12N9/10
CPCC12N9/1007C12P13/12
Inventor 由永峰
Owner 由永峰
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