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Immobilized lysine decarboxylase, preparation thereof, 1, 5-pentane diamine preparation method and product

A lysine decarboxylase, amine-based technology, applied in biochemical equipment and methods, immobilized on/in organic carriers, enzymes, etc., can solve problems such as low stability and low reuse rate

Active Publication Date: 2018-03-09
CATHAY R&D CENT CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In order to overcome the defects of low stability and low reuse rate of lysine decarboxylase in the field of biochemical production, the present invention provides the following technical solutions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0132] Preparation Example Preparation of Lysine Decarboxylase

[0133] The microbial glycerol preservation bacterium liquid of expressing lysine decarboxylase is inoculated in 500 milliliters of seeds that 100 milliliters of liquid culture medium (LB medium, comprise peptone 1%, yeast powder 0.5 percent, sodium chloride 1 percent, pH7.0) are housed Cultivate in the bottle at 37°C and 200rmp shaker for 15h to obtain seed solution. In a 5L fermenter, add 3 liters of LB medium, sterilize at 121°C for 20 minutes and then insert the above seed liquid to start fermentation (the formula of the fermentation medium is LB medium: peptone 1%, yeast powder 0.5%, sodium chloride 1 %, pH7.0), start fermentation at 30°C and 300 rpm, control the ventilation rate to 0.3vvm, and tank pressure to 0.04MPa; control the pH value of the fermentation broth to 7.0 during the fermentation process, and stop the fermentation after 25 hours of fermentation. After the fermentation, collect the cells (tha...

Embodiment 1

[0135] Modification of Polyvinyl Alcohol Carrier and Preparation of Immobilized Lysine Decarboxylase

[0136] (1) Take SESA (β-ethylsulfonyl aniline sulfate), add it to 2% sodium carbonate aqueous solution, add sodium hydroxide to a final concentration of 0.1mol / L, grind and dissolve, and after stabilization, use filter paper Filtrate, and take the clear liquid as etherified liquid, the concentration of etherified liquid is 20wt%.

[0137] (2) Take 100g of filamentous polyvinyl alcohol (PVA) with a degree of polymerization of 3000, add 300g of the etherification solution prepared in step (1), and carry out etherification reaction under the condition of heating in a water bath at 60°C, and add dropwise 4mol / L sodium hydroxide solution, stirred manually until the pH was about 11, stopped adding alkali, and continued stirring for 1 hour. After the reaction, the etherified polyvinyl alcohol was washed with hot alkaline water (0.1N NaOH) below 60°C, and then washed repeatedly with...

Embodiment 2

[0144] Modification of Polyvinyl Alcohol Carrier and Preparation of Immobilized Lysine Decarboxylase

[0145] (1) Take SESA (β-ethylsulfonyl aniline sulfate), add it to 2% sodium carbonate aqueous solution, add sodium hydroxide to a final concentration of 0.1mol / L, grind and dissolve, and after stabilization, use filter paper Filtrate, and take the clear liquid as etherified liquid, the concentration of etherified liquid is 10wt%.

[0146] (2) Take 100g of filamentous polyvinyl alcohol with a degree of polymerization of 3000, add 300g of the etherification solution prepared in step (1), and carry out etherification reaction under the condition of heating in a water bath at 60°C, and add 4mol / L dropwise at the same time The sodium hydroxide solution was manually stirred until the pH was about 11, the addition of alkali was stopped, and the stirring reaction was continued for 1 h. After the reaction, wash the etherified PVA (polyvinyl acetate) silk with hot alkaline water (0.1N...

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Abstract

The invention relates to immobilized lysine decarboxylase, a preparation thereof, a 1, 5-pentane diamine preparation method and a product. The invention specifically relates to immobilized lysine decarboxylase comprising a diazo group-modified polymer carrier and lysine decarboxylase, and a method for preparing immobilized lysine decarboxylase by the diazo group-modified polymer carrier; a methodfor preparing 1, 5-pentane diamine by the immobilized lysine decarboxylase, and the prepared 1, 5-pentane diamine thereof. The immobilization method has high enzyme immobilization efficiency and goodstability in use, improves the use efficiency of enzyme, solves the problem of poor use stability of free lysine decarboxylase (cells) in traditional processes, greatly reduces the use cost of the carrier and the biological production cost of 1, 5-pentanediamine, simplifies the separation step of 1, 5-pentane diamine solution and enzyme, increases the degree of automation of the production of 1, 5-pentane diamine, and promotes the process of industrialization of biological production of 1, 5-pentanediamine.

Description

technical field [0001] The present invention relates to an immobilized lysine decarboxylase and a preparation method thereof, a preparation method of 1,5-pentanediamine and the obtained 1,5-pentanediamine; in particular, it relates to a high Molecular carrier and immobilization of lysine decarboxylase lysine decarboxylase, and chemical modification of macromolecular substances to obtain diazo-modified macromolecular carriers, and preparation of immobilized diazo-modified macromolecule carriers The method of lysine decarboxylase. The invention also relates to a method for preparing 1,5-pentanediamine by using the immobilized lysine decarboxylase and the prepared 1,5-pentanediamine. Background technique [0002] Biocatalytic reactions have the characteristics of mild conditions, high selectivity, and high reaction efficiency, so they are increasingly used in the field of chemical synthesis. However, due to the poor stability of biological enzymes in in vitro industrial envir...

Claims

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Application Information

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IPC IPC(8): C12N11/12C12N11/08C12P13/00
CPCC12N9/88C12N11/08C12N11/12C12P13/001C12Y401/01018
Inventor 徐冠珠于丽珺刘修才
Owner CATHAY R&D CENT CO LTD
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