Method for preparing 1,5-pentanediamine

A technology for the fermentation of pentamethylenediamine and lysine, which is applied in directions such as fermentation to achieve the effects of simple process, cost saving and reduction of purification steps

Active Publication Date: 2015-07-08
CATHAY R&D CENT CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In summary, there are many studies on the catalysis of lysine to pentamethylenediamine by lysine decarboxylase produced by microorganisms, which is characterized by the use of purified lysine Amino acid or lysine salt, lysine needs to be purified, the process is complicated, the environment is polluted, and the production cost of pentamethylenediamine is high, which limits the industrial production of pentamethylenediamine

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  • Method for preparing 1,5-pentanediamine
  • Method for preparing 1,5-pentanediamine

Examples

Experimental program
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Effect test

preparation Embodiment 1

[0048] Preparation Example 1. Preparation of lysine fermentation stoste

[0049] Prepare lysine fermentation stock as follows

[0050] (1) Slope: use LB medium

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

[0052] (2) Seed cultivation

[0053] The corynebacterium slant with lysine production ability was inoculated in 200 ml of liquid medium (broth medium: beef extract 1%, peptone 1%, yeast extract 0.5%, sodium chloride 0.5%, pH 7.0 ) in a 500ml seed bottle at 33°C and 200rmp shaker for 10-15 hours.

[0054] (3) Lysine fermentation

[0055] In a 10L fermenter, add 5L of fermentation medium. Insert above-mentioned seed liquor and start to ferment. The fermentation medium of the fermenter contains: glucose 8%, molasses 1.2%, corn steep liquor 1%, ammonium sulfate 0.6%, dipotassium hydrogen phosphate 2%, magnesium sulfate heptahydrate 0.08%, iron sulfate heptahydrate 0.002%, sulfuric acid tetrahydrate Manganese 0.002%, the balanc...

preparation Embodiment 2

[0059] Preparation Example 2. Preparation of Lysine Decarboxylase Fermentation Broth

[0060] The lysine decarboxylase fermentation broth was prepared as follows:

[0061] (1) Seed cultivation

[0062] Inoculate the liquid medium (LB medium containing 1% of peptone, 0.5% of yeast powder, and 1% of sodium chloride) with 100 ml of liquid liquid culture medium (LB medium, containing 1% of peptone) by inoculating the glycerol-preserved bacterial liquid of haffney genetically engineered bacteria expressing lysine decarboxylase , pH7.0) in a 500ml seed bottle at 35°C, 170rmp shaker for 15 hours.

[0063] (2) Lysine decarboxylase fermentation

[0064] In a 5L fermenter, add 3 liters of LB medium, and start fermentation by inserting the above seed liquid after sterilizing at 121°C for 20 minutes (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 air flow rate to 0.3...

Embodiment 1

[0065] Embodiment 1. Lysine decarboxylase catalyzes lysine fermentation stoste

[0066] The lysine fermentation stoste that takes a certain amount of Preparation Example 1 method to obtain joins in the 500ml Erlenmeyer flask, and the lysine content is 11%; SO in the lysine fermentation stoste 4 2- The concentration is about 0.23mol / kg, which also contains sugar and free NH 4 + , total sugar content 350ppm, free NH 4 +The content is 0.032mol / kg. Add lysine decarboxylase cells (obtained by the method of Preparation Example 2, the same below) and coenzyme (5'-pyridoxal phosphate) to this lysine fermentation stoste, and shake the reaction for a certain period of time at a stirring speed of 200rpm, The reaction temperature was set at 38°C; the weight ratio of lysine decarboxylase in the reaction system to the weight ratio of lysine in the lysine fermentation stock solution, the coenzyme concentration (the ratio of the coenzyme added to the weight of all reaction materials exc...

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Abstract

The invention relates to a method for preparing 1,5-pentanediamine. The method comprises the step of adding lysine decarboxylase into a lysine fermentation broth to synthesize 1,5-pentanediamine. The method utilizes the lysine fermentation broth but not purified lysine as the substrate, so that the process is simpler and more economical.

Description

technical field [0001] The invention relates to a preparation method of 1,5-pentanediamine, in particular to a method for catalytically synthesizing 1,5-pentanediamine with lysine decarboxylase. Background technique [0002] 1,5-Diaminopentane (also known as 1,5-pentanediamine, referred to as pentamethylenediamine) is an important 5-carbon compound in the chemical industry. Its main purpose is to manufacture polyamide, polyurethane, etc., and it can also be used Manufacture of isocyanate, pyridine, piperidine and other important chemical raw materials. [0003] To date, diamines have been chemically produced from petroleum-based raw materials via dicarboxylic acid intermediates or by chemical decarboxylation of amino acids (Albrecht, Klaus et al.; Plastics; Winnacker-Kuechler (5th edition) (2005)). The rise in oil prices makes it an ideal way to synthesize diamines using renewable raw materials through bioengineering methods. [0004] There are two main methods of biosynt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00
Inventor 于丽珺刘驰秦兵兵李乃强
Owner CATHAY R&D CENT CO LTD
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