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Method for preparing L-ornithine-L-aspartate salt

A technology of aspartate, aspartic acid, applied in directions such as being fixed on/in organic carrier, fermentation, etc., can solve problems such as separation and purification of unfavorable products, increase in production cost, reduce product yield and the like, and achieve The effect of less impurities, low production cost and high yield

Inactive Publication Date: 2011-06-22
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-L-aspartate by enzymatic conversion, characterized in that the method comprises the steps of:

[0031] (1) Preparation of immobilized enzyme

[0032] The amount of raw materials is as follows: 50g of triethanolamine anionic styrene resin, 400mg of arginase, and saturated saline subject to being able to submerge and immobilize the enzyme, take 50g of triethanolamine anionic styrene resin, soak it 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, wherein the concentration of triethanolamine anion styrene resin is 40g / ml. Wash with cold 0.1M sodium bicarbonate until the pH value reaches 8.5, add 100ml of arginase solution (4g / l by weight), and react at 4°C for 24h. Wash with 0.1M sodium bicarbonate, 1M sodium chloride, and deionized water three times in sequence, 200ml each time, th...

Embodiment 2

[0038] (1) Preparation of immobilized enzyme

[0039] Take 40g of triethanolamine anionic styrene resin, soak in 1M potassium carbonate solution for 8h, filter and dry, add 10ml of 1M potassium carbonate solution, add 5ml of hydrogen bromide (1g / ml) under ice bath and react for 5min under ice bath , wash with cold 0.05M sodium bicarbonate until the pH value reaches 7, add 400ml of arginase solution (weight percent concentration is 1g / l) and react at 1°C for 12h. Wash with 100ml of 0.05M sodium bicarbonate solution, 100ml of 0.5M sodium chloride solution, and 100ml of deionized water for 2 times, and finally soak the immobilized enzyme in saturated saline for 0.5h, and wash until no chloride ion is detected in the solution. Obtain immobilized enzyme, store it in water, and set aside;

[0040] (2) Optimization of transformation conditions

[0041] Arginine 60g, L-aspartic acid 18.5g and manganese acetate 0.2g were mixed, dissolved in 240g deionized water to obtain a mixed solu...

Embodiment 3

[0045] (1) Preparation of immobilized enzyme

[0046] Take 50g of triethanolamine anionic styrene resin, soak in 2M potassium carbonate solution for 12h, filter and dry, add 100ml of 2M potassium carbonate solution, 10ml of hydrogen bromide (2g / ml) and react in ice bath for 15min, then use cold 0.1M Wash with cold sodium bicarbonate until the pH value reaches 8.5, add 100ml of arginase solution (4g / l by weight) 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 for 3 times, and finally soak the immobilized enzyme in saturated saline for 1 hour, and wash until no chloride ion is detected in the solution to prepare the immobilized enzyme. Catase, stored in water, for subsequent use;

[0047] (2) Optimization of transformation conditions

[0048] Arginine 60g, L-aspartic acid 20g and manganese acetate 0.2g are mixed, dissolve in 240g deionized water, the immobilized enzyme (wet) 50.4g...

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Abstract

The invention relates to a method for preparing L-ornithine-L-aspartate salt. The L-ornithine-L-aspartate salt is produced through conversion of arginase. The method comprises the following process steps of: (1) preparing immobilized enzyme; (2) optimizing conversion conditions; and (3) extracting product and refining. Compared with the prior art, the method has the advantages that: production cost is low, the production conditions are mild, impurities in a conversion system are a few, the process steps are simple, the production operation is safe, the purity is high, 300 to 320g of L-ornithine-L-aspartate salt is contained in each liter of reaction solution, the yield is high, the conversion rate of arginase is over 95 percent and the like.

Description

technical field [0001] The invention relates to a preparation method of amino acid salt, in particular to a preparation method of L-ornithine-L-aspartate. Background technique [0002] L-ornithine-L-aspartate is an important medicine and a nutritional supplement for liver cirrhosis, which can promote urea production, reduce blood ammonia, accelerate the tricarboxylic acid cycle, start intracellular respiration, and improve liver function. It is an effective drug for reducing blood ammonia and is used to treat hyperammonemia. [0003] At present, the preparation method for L-ornithine-L-aspartate is mainly to crystallize after the direct reaction of L-ornithine and L-aspartic acid, and the preparation of L-ornithine is to prepare L-ornithine Acid-L-Aspartate Key. [0004] At present, there are different 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 met...

Claims

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Application Information

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IPC IPC(8): C12P21/02C12N11/08
Inventor 苏建勇郑南华周会友
Owner 湖南天成生化科技有限公司
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