Method using thallus whole cells to catalyze arginine to convert into agmatine

A technology of agmatine and whole cells, which is applied in the field of bioengineering, can solve the problems of cell crushing, purification and extraction, and increase the burden, and achieve the effects of high conversion efficiency, easy crystallization and purification, and short fermentation cycle

Active Publication Date: 2019-09-20
JINAN GUOLI BIOLOGICAL SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese invention patent application CN104911223A discloses a biological preparation method of agmatine sulfate, although its yield can reach 200g / L, but it breaks the cells, which undoubtedly increases the burden on the subsequent purification and extraction

Method used

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  • Method using thallus whole cells to catalyze arginine to convert into agmatine
  • Method using thallus whole cells to catalyze arginine to convert into agmatine
  • Method using thallus whole cells to catalyze arginine to convert into agmatine

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Experimental program
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Effect test

Embodiment 1

[0037] Embodiment 1: Optimization of crystallization conditions of agmatine

[0038] Since the existing scientific research materials have not yet found relevant information on the crystallization of agmatine, the present invention investigates the influence of various organic solvents on the crystallization of agmatine, including methanol and ethanol. Although these organic solvents are all beneficial to the crystallization of agmatine to a certain extent, considering the safety factor of agmatine in application, the present invention focuses on the influence of ethanol on the crystallization of agmatine. Because low temperature is conducive to the precipitation of agmatine, the temperature was controlled at 4-15°C during the experiment, and the concentration of agmatine in the conversion supernatant was 200-300g / L, and concentrated in vacuum to 1 / 1 of the original volume. 2. Final result shows that the ethanol that adds 55% ratio is more conducive to the recovery of agmatin...

Embodiment 2

[0040] Example 2: A method for converting arginine into agmatine by catalyzing whole cells of bacteria

[0041] The method comprises the steps of:

[0042] (1) Preparation of whole-cell arginine decarboxylase concentrate

[0043] a. Construction of Arginine Decarboxylase Genetic Engineering Bacteria

[0044] The genetically engineered bacterium uses Escherichia coli K12 as the host, pET28A as the carrier, and adiA as the target gene. The nucleotide sequence of the adiA gene is shown in SEQ ID NO.1, which can be constructed by conventional molecular cloning techniques.

[0045] b. Arginine decarboxylase genetic engineering bacteria pilot test fermentation culture

[0046] the s 1 Take the glycerol tube of the engineering bacteria and inoculate it on the culture dish plate to activate the strains, and the culture time is 24 hours; store the culture dish plate in a refrigerator at 4°C; use an inoculation loop to dig out a ring of plate seeds under aseptic conditions and inocul...

Embodiment 3

[0059] Example 3: A method for converting arginine into agmatine by catalyzing whole cells of bacteria

[0060] The method comprises the steps of:

[0061] (1) Preparation of whole-cell arginine decarboxylase concentrate

[0062] a. Construction of Arginine Decarboxylase Genetic Engineering Bacteria

[0063] The genetically engineered bacterium uses Escherichia coli K12 as the host, pET28A as the carrier, and adiA as the target gene. The nucleotide sequence of the adiA gene is shown in SEQ ID NO.1, which can be constructed by conventional molecular cloning techniques.

[0064] b. Arginine decarboxylase genetic engineering bacteria pilot test fermentation culture

[0065] the s 1 Take the glycerol tube of the engineering bacteria and inoculate it on the culture dish plate to activate the strains, and the culture time is 24 hours; store the culture dish plate in a refrigerator at 4°C; use an inoculation loop to dig out a ring of plate seeds under aseptic conditions and inocul...

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Abstract

The method relates to a method using thallus whole cells to catalyze arginine to convert into agmatine and belongs to the technical field of bioengineering. The method includes: preparing whole-cell arginine decarboxylase concentrate, mixing the whole-cell arginine decarboxylase concentrate with initial conversion liquid, converting, crystalizing and recrystallizing. The method has the advantages that by controlling the carbon source, nitrogen source and dissolved oxygen during fermentation induction, the obtained arginine decarboxylase genetically engineered thallus whole-cell concentrate is high in arginine catalytic conversion efficiency; the concentrate does not contain byproducts, so that the agmatine obtained by conversion can be conveniently crystalized and purified; the thallus whole-cell concentrate is short in fermentation period, a large amount of the thallus whole-cell concentrate can be obtained in a short time, and large-scale preparation of the agmatine is benefited; agmatine product concentration on conversion liquid is 200-300g / L, conversion rate is 92-99%, agmatine product purity can reach 97-100%, and product yield can reach 90-100%.

Description

technical field [0001] The invention relates to a method for converting arginine into agmatine by catalyzing whole cells of bacteria, belonging to the technical field of bioengineering. Background technique [0002] As an important biogenic amine, agmatine has great physiological functions and medical value. At present, the preparation of agmatine in my country mainly comes from chemical methods. The chemical synthesis of agmatine has cumbersome steps, low efficiency, high cost, heavy pollution, and does not meet the requirements of environmental protection. The use of biological methods to synthesize agmatine has attracted people's attention because of its advantages of fast reaction rate, simple operation, and no pollution. [0003] Agmatine is produced by the decarboxylation of the substrate arginine catalyzed by arginine decarboxylase (see figure 1 ). The concentration of agmatine in the blood is 0.2-0.4ng / ml, and agmatine is distributed in almost all organs in the bo...

Claims

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Application Information

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IPC IPC(8): C12P13/00C12N9/88C12R1/19
CPCC12N9/88C12P13/001C12Y401/01019
Inventor 袁建国张言慧高先岭
Owner JINAN GUOLI BIOLOGICAL SCI & TECH
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