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Method for separating L-ornithine from L-ornithine conversion solution prepared by using enzyme biotechnology and forming L-ornithine hydrochloride

A technology of ornithine hydrochloride and biotechnology, which is applied in the field of separating ornithine and forming ornithine hydrochloride, which can solve the problems of increasing production costs due to production requirements, increasing production costs of enterprises, and reducing the technical threshold Effect

Active Publication Date: 2014-12-10
WUXI JINGHAI AMINO ACID
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The process diagram of domestic enterprises using enzymatic method to prepare L-ornithine is as attached figure 1 As shown, two disadvantages of the existing technology are found from the figure: 1, the extraction of L-ornithine needs to be carried out in an explosion-proof workshop, because absolute ethanol is used in the extraction process, which increases the production requirements and Increased production costs; 2. The waste liquid (including ethanol + urea) generated during the extraction process is a hazardous waste liquid that must be specially treated by a qualified company, which significantly increases the production cost of the enterprise

Method used

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  • Method for separating L-ornithine from L-ornithine conversion solution prepared by using enzyme biotechnology and forming L-ornithine hydrochloride
  • Method for separating L-ornithine from L-ornithine conversion solution prepared by using enzyme biotechnology and forming L-ornithine hydrochloride
  • Method for separating L-ornithine from L-ornithine conversion solution prepared by using enzyme biotechnology and forming L-ornithine hydrochloride

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Experimental program
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Effect test

Embodiment 1

[0025] The method steps of separating ornithine and forming ornithine hydrochloride from the ornithine conversion liquid prepared by enzyme biotechnology of the present invention are as follows:

[0026] 1. Preparation of arginase solution:

[0027] (1) Seed liquid culture:

[0028] Inoculate the arginase-producing Escherichia coli engineering bacteria E.coli BL21 in 500mL of LB liquid medium, and cultivate it to OD at 37°C and 200rpm 600 The value is 2.71, spare.

[0029] (2) Fermentation culture:

[0030] The seed liquid was added in a ratio of 1:100 (v / v), that is, 50 L of LB liquid medium was added to a 70 L fermenter, and then 0.5 L of seed liquid was added for fermentation. The culture temperature was 37°C, the rotational speed was 300 rpm, and the initial pH was 7.2. Sampling every 1 hour, when OD 600 When the value is about 4.0, the tank is lowered and the bacteria are collected.

[0031] (3) The fermented liquid in the lower tank first passes through a 200-mesh ...

Embodiment 2

[0051] The difference between this embodiment and embodiment 1 is:

[0052] Step 3: Separating ornithine in the ornithine conversion solution and forming a salt

[0053] Take 100L of the conversion solution and separate it with nanofiltration membrane 1, the molecular weight cut off of the nanofiltration membrane 1 is 300Da. Thereby obtain filtrate 1; Then filtrate 1 is filtered with nanofiltration membrane 2, and the molecular weight cut-off of this nanofiltration membrane 2 is 100Da, also obtained urea solution simultaneously, and filtrate 2 is adjusted pH5.0-6.0 with hydrochloric acid, forms ornithine For the hydrochloride solution, use 767 activated carbon to decolorize and absorb the trace impurities remaining in ornithine hydrochloride. The decolorization temperature is 80°C, and the stirring is continued for 30 minutes. The amount of activated carbon added is 20% of the solid content of the ornithine hydrochloride solution. , decarburization to obtain filtrate 3; above...

Embodiment 3

[0058] The difference between this embodiment and embodiment 1 is:

[0059] Step 3: Separating ornithine in the ornithine conversion solution and forming a salt

[0060] Take 100L of the conversion solution and separate it with nanofiltration membrane 1, the molecular weight cut off of the nanofiltration membrane 1 is 300Da. Thereby obtain filtrate 1; Then filtrate 1 is filtered with nanofiltration membrane 2, the molecular weight cut-off of this nanofiltration membrane 2 is 60Da, obtains filtrate 2, also obtained urea solution simultaneously, filtrate 2 is regulated PH5.0-6.0 with hydrochloric acid, Form ornithine hydrochloride solution, use 767 type activated carbon to decolorize and absorb trace impurities remaining in ornithine hydrochloride, decolorize temperature 80°C, keep stirring for 30min, the amount of activated carbon added is the solid content of ornithine hydrochloride solution 20% of content, decarburization obtains filtrate 3; above-mentioned filtrate 3 is sub...

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Abstract

The invention provides a method for separating L-ornithine from an L-ornithine conversion solution prepared by using an enzyme biotechnology and forming L-ornithine hydrochloride. The L-ornithine conversion solution is obtained by converting arginase into arginine. The method comprises the concrete steps of firstly, treating the conversion solution by using a nanofiltration membrane 1 with the molecular weight cutoff of 300-1000Da to obtain filtrate 1; then, treating the filtrate 1 by using a nanofiltration membrane 2 with the molecular weight cutoff of 50-100Da to obtain filtrate 2 containing L-ornithine; and after adding acid to form salt, decoloring by using activated carbon, and separating a crystal to obtain the L-ornithine hydrochloride. The traditional ethanol extraction method is replaced with the method provided by the invention, so that the L-ornithine hydrochloride is extracted without absolute ethyl alcohol, furthermore, an anti-explosion workshop does not need to be built by a production enterprise of the L-ornithine hydrochloride, the technical threshold for producing the L-ornithine hydrochloride is reduced, and the method is also safe and reliable; in addition, a urea solution generated in the process that the L-ornithine hydrochloride is prepared by using the method provided by the invention can be used as a fertilizer to be directly used for agricultural production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for separating ornithine from ornithine conversion liquid prepared by enzyme biotechnology and forming ornithine hydrochloride. Background technique [0002] Ornithine was discovered by Jeff in the hydrolyzate of bird urine in 1877, so it was named ornithine. L-Ornithine is a non-protein amino acid in free form. In organisms, it mainly participates in the urea cycle: ornithine combines with a molecule of ammonia and a molecule of carbon dioxide to form citrulline, which then reacts with a molecule of ammonia to form arginine, which is hydrolyzed by arginase. A molecule of urea and a molecule of ornithine are produced, so that the ammonia in the body is repeatedly excreted. Ornithine is a kind of diaminovaleric acid, which has D-form and L-form, and mostly exists in L-form. [0003] Ornithine is very important to the liver. The protective function of ornithine ...

Claims

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Application Information

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IPC IPC(8): C07C227/40C07C227/18C07C229/26C07C273/16
Inventor 宁健飞侯一鸣蔡立明
Owner WUXI JINGHAI AMINO ACID
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