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Method of preparing cadaverine by virtue of immobilized hafnia alvei

A technology of Hafnia haffonia and cadaverine, which is applied in the field of immobilized Hafnia haffonia to produce cadaverine, can solve the problems of difficult separation of bacteria and fermentation broth, low conversion efficiency of cadaverine, and low concentration of fermentation strains. Achieve the effect of shortening fermentation cycle, increasing stability and reducing production cost

Inactive Publication Date: 2014-04-16
天津北洋百川生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional fermentation method uses Hafnia hives to produce cadaverine, because of the disadvantages of low concentration of fermentation strains, low tolerance of strains to substrate concentration, low activity of LDC, and difficulty in separating bacteria and fermentation broth after fermentation transformation, etc. Make the transformation efficiency of cadaverine low, increase the production cost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for producing cadaverine by embedding Hafnia alvei, the steps are as follows:

[0026] 1. Activate the Hafnia alvei AS1.1009 strain for 6-8 hours, that is, take 1 ring of bacteria from the slant medium and transfer it into a 500mL Erlenmeyer flask containing 50mL seed medium, at 35°C, Shake culture at 170r / min for 12h to obtain seed solution. The composition of the seed medium is (g / L): peptone 10, beef extract 5, NaCl 5, corn steep liquor 5, pH 7.2.

[0027] 2. Put the seed liquid obtained in step 2 into a 5L (3L liquid) fermenter at a volume ratio of 5%, at 35°C, with a ventilation rate (v / v) 1:1.5, and a stirring speed of 300-600rpm , dissolved oxygen 20%, cultivated for 24h. The composition of the fermentation medium is (g / L): glucose 20, corn steep liquor 20, yeast extract 5, beef extract 3, peptone 10, NaCl5, ammonium sulfate 2, vitamin B 6 1, pH7.0.

[0028] 3. Strain embedding and fixing: Weigh 68g of sodium alginate and add it to 1000mL of distille...

Embodiment 2

[0031] 1, the preparation of seed liquid and the bacterial classification used are with embodiment 1.

[0032] 2. The preparation of the fermented liquid is the same as in Example 1.

[0033] 3. Strain embedding and fixation: Weigh 70g of sodium alginate and add it to 1000mL distilled water, then add 2g of polyvinyl alcohol and mix evenly, put it in an autoclave at 121°C, sterilize for 30min, and cool it in an ultra-clean bench to Gel formed at room temperature. Centrifuge 500mL of Hafnia alvei fermentation broth to collect the bacteria, then resuspend it with 10mL of normal saline and add it to the gel, stir and mix evenly, add dropwise to 2% sterilized CaCl with a sterile 10mL syringe 2In the solution, after the dropwise addition, place it in a clean bench at room temperature to continue cross-linking for 10 hours to form immobilized pellets, and then wash with sterilized normal saline for 3 times to obtain the embedded microbial agent. All the above operations must be per...

Embodiment 3

[0036] 1, the preparation of seed liquid and the bacterial classification used are with embodiment 1.

[0037] 2. The preparation of the fermented liquid is the same as in Example 1.

[0038] 3. Fermentation tank seed solution embedding: Weigh 75g of sodium alginate into 1000mL of distilled water, then add 4g of polyvinyl alcohol and mix evenly, put it in an autoclave at 121°C, sterilize for 30min, and cool it in an ultra-clean bench to room temperature to form a gel. Centrifuge 500mL of Hafnia alvei fermentation broth to collect the bacteria, then resuspend it with 10mL of normal saline and add it to the gel, stir and mix evenly, add dropwise to 2% sterilized CaCl with a sterile 10mL syringe 2 In the solution, after the dropwise addition, place it in a clean bench at room temperature to continue cross-linking for 10 hours to form immobilized pellets, and then wash with sterilized normal saline for 3 times to obtain the embedded microbial agent. All the above operations must...

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Abstract

The invention relates to a method of preparing cadaverine by virtue of immobilized hafnia alvei (Hafnia Alvei) AS1.1009, belongs to the field of microbial fermentation and aims to increase the production rate of the cadaverine by utilizing the microbial embedding technology. The method comprises the steps: (1) carrying out third-stage culturing on a hafnia alvei strain; (2) embedding and fixing a bacteria solution by utilizing an embedding carrier under the aseptic condition; and (3) adding an immobilized thallus into a fermentation tank, and carrying out fermentation conversion to prepare the cadaverine, wherein the conversion rate of the cadaverine after the fermentation reaches 95.31-97.42 percent. With the adoption of the embedded high-concentration hafnia alvei, a strain microorganism with high concentration is provided; and the stability of the strain and the tolerance of the concentration of a substrate are improved by embedding microorganism particles, the enzymatic activity is improved, and the productivity of the cadaverine is increased.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to a method for producing cadaverine by immobilized Hafnia hives. Background technique [0002] Cadaverine, that is, 1,5-diaminopentane, belongs to the diamines in polyamines, and is one of the components in the odor of corpse decay. The five-carbon compound 1,5-pentanediamine (cadaverine) has received increasing attention as a bio-based platform compound, especially as a building monomer for novel high-performance bio-based polymers. Research on its production strains and production process, and the huge economic benefits that can be brought by the supply of bio-based 1,5-pentanediamine and its derivatives have also aroused great interest. L-lysine decarboxylase (L-lysinedecarboxylase, referred to as LDC, EC4.1.1.15), is a reversible decoction of lysine 2 An enzyme that generates cadaverine (1,5-pentanediamine), an inducible intracellular enzyme, needs pyridoxal ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12R1/01
Inventor 乔长晟彭巧林青石漫漫
Owner 天津北洋百川生物技术有限公司
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