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A method for separating and extracting 2-ketobutyric acid from enzyme conversion solution

A technology for ketobutyric acid and enzymatic conversion, applied in the separation/purification of carboxylic acid compounds, organic chemistry, etc., can solve the problems of high production cost, low yield, low product purity, etc., to improve yield, avoid volatilization, good effect

Inactive Publication Date: 2017-07-11
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a novel separation and extraction process of 2-ketobutyric acid, which can change the problems of complex separation and extraction process, low yield, low product purity and high production cost in the existing production process.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] a. Take 5.0L of 2-ketobutyrate conversion solution (-ketobutyric acid content 42.3g / L), adjust the pH to 4.0 with hydrochloric acid, enter the microfiltration membrane separation system for microfiltration, and obtain the -ketobutyrate that removes the bacteria Acid microfiltrate; the microfiltration membrane separation system adopts a hollow fiber membrane separation system, the microfiltration membrane pore size is 0.2um, the operating temperature is 35°C, the pressure difference between the inlet membrane and the outlet membrane is 0.1Mpa, and the filtration process is divided into several times to add Top wash with purified water until the 2-ketobutyric acid content in the dope is lower than 2.0g / L, then shut down to obtain the 2-ketobutyric acid microfiltrate. The recovery rate of supernatant 2-ketobutyric acid reaches 96.2%, the cell removal rate is greater than 99.9%, and the membrane flux reaches 89L·(m 2 h) -1 .

[0024] b. Pump the clarified 2-ketobutyric ac...

Embodiment 2

[0031] Get 2-ketobutyrate enzyme conversion solution 6.8L (2-ketobutyric acid content 35.3g / L), regulate pH with hydrochloric acid and be 4.0, carry out microfiltration and ultrafiltration to remove protein according to the method for embodiment 1 (wherein , microfiltration conditions: the microfiltration membrane pore size is 0.1um, the temperature is 50 ° C, and the pressure difference between the inlet and outlet membranes is 0.05 MPa; the ultrafiltration conditions: the temperature is 20 ° C, the operating pressure difference is 0.05 MPa), and the weight of the ultrafiltrate is 1.5 % activated carbon for decolorization, temperature 45°C, stirring and decolorizing for 25min, filtering to obtain 2-ketobutyric acid decolorization solution, adjusting the pH to 11.0 with sodium hydroxide, adding 1.5% of the weight of the decolorization solution by activated carbon to adsorb flocculent protein, and then decompressing Evaporate to 18% of the original volume liquid to obtain 2-keto...

Embodiment 3

[0033] Get 8.4L of 2-ketobutyrate conversion solution (2-ketobutyric acid content 37.2g / L), adjust pH with hydrochloric acid to be 4.0, carry out microfiltration and ultrafiltration to remove protein according to the method for embodiment 1 (wherein , microfiltration conditions: the microfiltration membrane pore size is 0.1um, the temperature is 10°C, and the pressure difference between the inlet and outlet membranes is 0.1Mpa; the ultrafiltration conditions: the temperature is 50°C, the operating pressure difference is 0.08MPa), and the weight of the ultrafiltrate is 1.8 % activated carbon for decolorization, temperature 40°C, stirring and decolorizing for 30min, filtering to obtain 2-ketobutyric acid decolorization solution, adjusting the pH to 11.0 with sodium hydroxide, adding 1.8% of the weight of the decolorization solution by activated carbon to adsorb flocculent protein, and then decompressing Evaporate to 20% of the original volume liquid to obtain 2-ketobutyric acid c...

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Abstract

Provided is a method for extracting 2-ketobutyric acid from an enzymatic conversion solution. When an enzyme catalysis method is adopted to produce 2-ketobutyric acid, an enzyme catalysis solution also contains thallus, water-soluble protein, pigment, threonine, sodium salt and other impurities besides the 2-ketobutyric acid, and due to the fact that the 2-ketonebutyric acid has the characteristics of easy dissolution, low boiling point and the like in a water phase and an organic phase, separation and extraction are difficult. According to the method, first system cross flows are separated and the thallus, most of the protein and the pigment filtered and removed through double membranes, namely a micro-filtration membrane and an ultra-filtration membrane, a membrane filtration solution is sequentially subjected to the operation steps of decoloring by means of powdered activated carbon, alkalified protein removal, reduced-pressure concentration, impurity removal by means of a cation exchange column and frozen vacuum drying, and 2-ketone butyric acid crystals with the purity above 95.0% are obtained, and the yield is above 80%. The method adopts the micro-filtration and ultra-filtration membranes to filter the thallus and the protein, the thallus is removed thoroughly, and a filtration solution is clear and transparent; the impurities are removed by adopting a cation exchange process, and the threonine, sodium salt and other impurities difficult to separate are efficiently removed.

Description

technical field [0001] The invention relates to a method for separating and extracting 2-ketobutyric acid from enzyme conversion liquid, and belongs to the technical field of organic acid production by fermentation. Background technique [0002] 2-ketobutyric acid is also known as a-ketobutyric acid (a-Ketobutyric acid), 2-oxobutyric acid (2-Oxobutyric acid), 3-methylpyruvate, the molecular formula is C 4 h 6 o 3 , with a relative molecular mass of 102.09, a melting point of 31-32°C, and a boiling point of 82-84°C (16mmHg). It is easily soluble in water and alcohol, and has a sweet, roasted, creamy, caramel and fat flavor. It is a precursor substance for the synthesis of many useful compounds, such as isoleucine, 1-propanol, n-butanol, 2-hydroxybutyric acid, baolomycin, furanone and some food additives, etc., especially, 2- Ketobutyric acid can synthesize L-homoalanine, which is an important precursor of antiepileptic drugs levetiracetam (LEV) and buvaracetam tablets. An...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C51/42C07C51/47C07C59/347
CPCC07C51/42C07C51/47C07C59/347
Inventor 陈宁齐俊生徐庆阳范晓光谢希贤张成林李燕军
Owner TIANJIN UNIV OF SCI & TECH
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