Preparation process of 1,5-pentamethylene diamine

A technology of pentamethylenediamine and seeds, which is applied in the field of preparation of 1,5-pentamethylenediamine, can solve the problems of short fermentation period, toxicity of pentamethylenediamine microbial strains, etc., and achieve short production period, low cost and reduced toxicity impact Effect

Pending Publication Date: 2019-03-29
CATHAY R&D CENT CO LTD +1
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Problems solved by technology

[0005] In order to overcome the above technical deficiencies, the present invention provides a method of mixed fermentation to solve the problem of the toxicity of pentamethylenediamine to microbial strains. The process is simple, the cost is low, and the fermentation period is short. The preparation method of diamine

Method used

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  • Preparation process of 1,5-pentamethylene diamine
  • Preparation process of 1,5-pentamethylene diamine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1 prepares lysine fermented liquid

[0040] (1) Inclined surface: use LB medium

[0041] LB medium: peptone 1%, yeast extract 1%, sodium chloride 0.5%, pH 7.2.

[0042] (2) Primary seed medium (shake flask seed medium)

[0043] The lysine producing strain used is Escherichia coli (Escherichia coli) ELa6116 (preservation number: CCTCCNO: M 2018736).

[0044] Get a ring of Escherichia coli (Escherichia coli) ELA6116 slant bacterial classification, inoculate in 100 milliliters of liquid culture medium (broth culture medium: beef extract 1%, peptone 1%, yeast extract 0.5%, sodium chloride 0.5%, pH Value 7.0) in the 500 milliliter seed bottle, at 33 ℃, 200rmp shaker culture 15-20 hour, obtain lysine primary seed liquid.

[0045] (3) Lysine fermentation conditions

[0046] In a 500mL Erlenmeyer flask, add 100mL of fermentation medium, insert 4mL of the above-mentioned lysine primary seed solution, and start fermentation at a temperature of 33°C and a shaker spe...

Embodiment 2

[0048] Embodiment 2 prepares the seed liquor of producing lysine decarboxylase bacterial strain

[0049] The strain used: Hafnia alvei strain Am607 (deposit number: CCTCC NO: M2018737).

[0050] Method: Take 200uL of the glycerol-preserved bacterial solution of the strain, inoculate it into a 500mL seed bottle filled with 100mL liquid seed medium, and culture it on a shaker at 35°C and 170rmp for 15 hours to obtain the seed solution of the lysine decarboxylase-producing strain. After testing, the content of lysine decarboxylase in the seed liquid is no more than 20U / mL.

[0051] LB medium: peptone 1%, yeast powder 0.5%, sodium chloride 1%, pH 7.0.

[0052] If no special instructions, following examples and comparative example adopt embodiment 1,2 method to prepare lysine fermented liquid and produce the seed liquid of lysine decarboxylase bacterial strain.

Embodiment 3

[0054] Take 1mL of the seed liquid of the lysine decarboxylase-producing strains cultured for 15 hours, respectively, and transfer them into the lysine fermentation broth fermented to 20h, 30h, 40h, 50h, 60h, 70h and 80h, without controlling the pH value of the fermentation process. Shake culture at 200rpm at 35°C. When the content of pentamethylenediamine no longer increases, it is judged that the fermentation is over. The actual incubation time is 20h, and the content of pentamethylenediamine in the fermentation broth is measured at the end of the fermentation. The results are shown in Table 1.

[0055] Table 1

[0056]

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Abstract

The present invention relates to a preparation process of 1,5-pentamethylene diamine. According to the method, a seed liquid containing lysine decarboxylase strains is introduced during lysine fermentation, and mixed fermentation is carried out. The method is suitable for industrial production of 1,5-pentamethylene diamine, minimizes toxicity of pentamethylene diamine to microorganisms to avoid adverse effects on fermentation, makes the subsequent process more simplified, and has certain commercial value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of 1,5-pentanediamine. Background technique [0002] 1,5-Diaminopentane (also known as 1,5-pentanediamine, referred to as pentamethylenediamine) is an important 5-carbon platform compound in the chemical industry. Its main purpose is to manufacture polyamide, polyurethane, etc., and it can also be used To manufacture isocyanate, pyridine, piperidine and other important chemical raw materials. [0003] There are two main methods of biosynthesizing pentamethylenediamine. One is that microorganisms use glucose to synthesize pentamethylenediamine through complex metabolic regulation. This method is complex in metabolic regulation, and pentamethylenediamine is toxic to microorganisms, which affects production efficiency. In 2007, Takashi et al. reported that by overexpressing lysine decarboxylase in Corynebacterium glutamicum with high lysine synthesis ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12R1/19C12R1/01
CPCC12P13/001
Inventor 路飞李乃强于丽珺周豪宏刘修才
Owner CATHAY R&D CENT CO LTD
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