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Preparation method for L-ornithine-alpha-ketoglutarate

A technology of ketoglutarate and ornithine, which is applied in the direction of fixing on/in organic carriers, fermentation, etc., can solve the problems of increased production cost, reduced product yield, separation and purification of unfavorable products, and achieves production The effect of low cost, less impurities and safe production operation

Inactive Publication Date: 2012-03-14
湖南天成生化科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Because the above-mentioned L-ornithinase production process uses free cells to transform the substrate L-arginine, so the transformation liquid will contain a small amount of bacterial protein and other impurities, which is not conducive to the separation and purification of the product; and the transformation must be removed. Bacteria, reducing product yield, resulting in increased production costs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A method for producing L-ornithine-α-ketoglutarate by enzymatic conversion is characterized in that the method comprises the following steps:

[0031] (1) Preparation of immobilized enzyme

[0032] The amount of raw materials is as follows: 50g of triethanolamine anion styrene resin, 400mg of arginase, saturated saline water as long as it can immerse the immobilized enzyme, take 50g of triethanolamine anion styrene resin, soak in 2M potassium carbonate solution for 12h, filter, After filtering to dryness, add 100ml 2M potassium carbonate solution, add 10ml hydrogen bromide (2g / ml) under ice bath and react for 15min under ice bath, in which the concentration of triethanolamine anion styrene resin is 40g / ml. Wash with cold 0.1M sodium bicarbonate to pH 8.5, add 100ml of arginase solution (weight percentage 4g / l), react at 4°C for 24h. Wash with 0.1M sodium bicarbonate, 1M sodium chloride, deionized water 3 times, 200ml each time, and then soak the immobilized enzyme in satura...

Embodiment 2

[0038] (1) Preparation of immobilized enzyme

[0039] Take 40g of triethanolamine anion styrene resin, soak it in 1M potassium carbonate solution for 8h, filter and filter dry, add 10ml 1M potassium carbonate solution, add 5ml hydrogen bromide (1g / ml) under ice bath and react for 5min under ice bath , Wash with cold 0.05M sodium bicarbonate to pH 7, add 400ml arginase solution (weight percent concentration is 1g / l) and react at 1°C for 12h. Wash twice with 0.05M sodium bicarbonate solution 100ml, 0.5M sodium chloride solution 100ml, 100ml deionized water, and finally soak the immobilized enzyme in saturated saline for 0.5h, wash until no chloride ions can be detected in the solution. Obtain the immobilized enzyme, store it in water for later use;

[0040] (2) Optimization of conversion conditions

[0041] Mix 60 g of arginine, 6.4 g of acetic acid and 0.2 g of manganese acetate, and dissolve them in 90 g of deionized water to obtain a mixed solution. Add 40.5 g of the wet immobiliz...

Embodiment 3

[0045] (1) Preparation of immobilized enzyme

[0046] Take 50g of triethanolamine anion styrene resin, soak it in 2M potassium carbonate solution for 12h, filter and filter dry, add 100ml 2M potassium carbonate solution, 10ml hydrogen bromide (2g / ml) and react for 15min in ice bath, with cold 0.1M Wash with cold sodium bicarbonate until the pH value reaches 8.5, add 100ml of arginase solution (weight percentage 4g / l) and react at 4°C for 24h. Wash with 200ml of 0.1M sodium bicarbonate solution, 200ml of 1M sodium chloride solution, and 200ml of deionized water successively for 3 times, and finally soak the immobilized enzyme in saturated saline for 1h, and wash until no chloride ion is detected in the solution. Chemical enzymes, stored in water for later use;

[0047] (2) Optimization of conversion conditions

[0048] Mix 60 g of arginine, 7 g of acetic acid and 0.2 g of manganese acetate, and dissolve them in 90 g of deionized water. Add 50.4 g of the immobilized enzyme (wet) prep...

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Abstract

The present invention relates to a preparation method for L-ornithine-alpha-ketoglutarate. The L-ornithine-alpha-ketoglutarate is produced by arginase transformation. The preparation method comprises the following steps: (1) preparation of immobilized enzyme; (2) optimization of transformation conditions; (3) product extraction and refining process. Compared to the prior art, the preparation method of the present invention has the following advantages that: the preparation method of the present invention has characteristics of low production cost, mild production conditions, less impurities in the transformation system, simple process steps, safe production operation and high purity; qualified products meeting the light transmission requirements can be synthesized; each liter of the reaction solution contains 300-320 g of the L-ornithine-alpha-ketoglutarate, and the transformation efficiency of the alpha-ketoglutaric acid is more than 90%; the preparation method further has other advantages.

Description

Technical field [0001] The invention relates to a preparation method of amino acid salt, in particular to a preparation method of L-ornithine-α-ketoglutarate. Background technique [0002] L-ornithine-α-ketoglutarate is a raw material widely used in medicine and health care. It is a muscle energy mixture. It can also help accelerate the growth of children with growth hormone deficiency and accelerate the wound healing of trauma patients. [0003] The method for preparing L-ornithine-α-ketoglutarate is mainly to crystallize after the direct salt reaction between L-ornithine and α-ketoglutarate. However, due to the different preparation methods of L-ornithine, L-ornithine-α-ketoglutarate can be prepared in different ways. [0004] At present, there are many reports on the preparation of L-ornithine at home and abroad. There are three main methods for preparing L-ornithine: chemical method, fermentation method, and enzymatic hydrolysis. [0005] Chemical method: The most commonly used p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/02C12N11/02
Inventor 苏建勇郑南华张建宇
Owner 湖南天成生化科技有限公司
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