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A method for separating ornithine and forming ornithine hydrochloride from ornithine conversion liquid prepared by enzyme biotechnology

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: 2016-05-04
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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  • A method for separating ornithine and forming ornithine hydrochloride from ornithine conversion liquid prepared by enzyme biotechnology
  • A method for separating ornithine and forming ornithine hydrochloride from ornithine conversion liquid prepared by enzyme biotechnology

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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 arginase-producing Escherichia coli engineering bacteria E.coliBL21 into 500mL 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 solution 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 solution was added for fermentation. The culture temperature is 37° C., the rotation speed is 300 rpm, and the initial pH is 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 dryin...

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 ornithine and forming ornithine hydrochloride from ornithine conversion liquid prepared by enzyme biotechnology, and the ornithine conversion liquid is obtained by using arginase to convert arginine , first using a molecular weight cut-off of 300-1000Da nanofiltration membrane 1 to process the conversion liquid to obtain a filtrate 1; then using a molecular weight cut-off of 50-100Da to process the filtrate 2 to obtain a filtrate 2 containing ornithine; After the acid is formed into a salt, it is decolorized with activated carbon and separated by crystallization to obtain ornithine hydrochloride. The method of the present invention replaces the traditional ethanol extraction method, so that the extraction of L-ornithine salt does not need to use absolute ethanol, so that the production enterprise of L-ornithine salt does not need to build an explosion-proof workshop, which reduces the production technology. The threshold also makes the method of the present invention safe and reliable; in addition, the urea solution produced in the process of preparing L-ornithine salt using the inventive method can be directly used in agricultural production as chemical fertilizer.

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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Patent Type & Authority Patents(China)
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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