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Method for domestication of 1,3-propanediol producing strain and use thereof

A technology for producing propylene glycol and bacteria, applied in the field of domestication 1, to achieve the effects of improving tolerance, increasing yield, and good application prospects

Inactive Publication Date: 2009-01-21
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Although the above methods can improve the tolerance of 1,3-propanediol-producing bacteria to substrates and / or certain products, in the actual fermentation process, cell growth is subject to the synergistic inhibition of substrates and various products, and these methods do not It cannot fundamentally solve the toxic effect of multiple product accumulation on cells and the huge negative impact on the production of 1,3-propanediol

Method used

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  • Method for domestication of 1,3-propanediol producing strain and use thereof
  • Method for domestication of 1,3-propanediol producing strain and use thereof
  • Method for domestication of 1,3-propanediol producing strain and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The domestication of embodiment 1, bacterial classification

[0028] The composition of the initial fermentation medium is (1L): K 2 HPO 4 4.45g, KH 2 PO 4 1.3g, (NH 4 ) 2 SO 4 2.0g, MgSO 4 ·7H 2 O 0.2g, CaCl 2 2H 2 O 0.02g, yeast powder 1.0g, glycerin 20g, iron solution 1.0mL, trace element solution 1.0mL, the balance is water. pH 7.0. Sterilize at 115°C for 20 minutes.

[0029] The composition of the above-mentioned iron solution is (1L): FeSO 4 ·7H 2 O 5g, 37% (volume percentage) HCl 4mL, the balance is water.

[0030] The composition of above-mentioned trace element solution is (1L): ZnCl 2 70mg, CuCl 2 2H 2 O 20mg, MnCl 2 4H 2 O0.1g, NiCl 2 ·6H 2 O 25 mg, H 3 BO 3 60 mg, Na 2 MO 4 2H 2 O 35mg, CoCl 2 2H 2 O 0.2g, 37% (volume percentage) HCl 4mL, the balance is water.

[0031] The domestication method of 1,3-propanediol producing bacteria specifically comprises the following steps:

[0032]1) Klebsiella K.oxytoca M5a1 (Ohta K, Beall...

Embodiment 2

[0039] Embodiment 2, fermentation effect contrast

[0040] Klebsiella K.oxytoca M5a1, K.pneumoniae AS1.1736, K.pneumoniae ATCC25955 and 24 strains of domesticated bacteria obtained from the domestication of the above-mentioned Example 1 were respectively inoculated into test tubes containing 5 mL of the above-mentioned initial fermentation medium, 37 Cultivate for 8 hours under the conditions of 180 r / min and a rotation radius of 12 mm to obtain seed liquid.

[0041] The above-mentioned seed solution was inserted into a 500-mL Erlenmeyer flask containing 100 mL of the above-mentioned initial fermentation medium according to the inoculation amount of 1% (volume percentage content), and cultured for 14 hours at 37 ° C, 180 r / m, and a radius of rotation of 12 mm. During the fermentation process, samples were taken at different time points to determine the OD of the fermentation broth 600nm value and concentration of 1,3-propanediol.

[0042] In the fermentation broth of domesti...

Embodiment 3

[0050] Embodiment 3, the domestication of bacterial classification

[0051] The composition of the initial fermentation medium is the same as in Example 1.

[0052] The domestication method of 1,3-propanediol producing bacteria specifically comprises the following steps:

[0053] 1) Klebsiella K.oxytoca M5a1 (Ohta K, Beall DS, Mejia JP, et al. Metabolic engineering of Klebsiella oxytoca M5A1 for ethanol production from xylose and glucose. Appl Environ Microbiol 1991, 157: 2810-2815.) (Tsinghua University), K.pneumoniae AS1.1736 (China General Microorganism Culture Collection Center CGMCC) and K.pneumoniae ATCC 25955 (American Culture Collection Center ATCC) were inoculated into the above-mentioned initial fermentation medium, 36 ° C 170r / m Under the condition that the radius of rotation is 11 mm, shake culture for 12 hours, centrifuge the fermentation broth to remove bacteria, and collect the supernatant. Glycerol was added to the supernatant so that the final concentration ...

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Abstract

The invention discloses a method for acclimating a 1, 3-propylene glycol producing strain and application of the 1, 3-propylene glycol producing strain. The method improves the tolerance of the 1, 3-propylene glycol producing strain on substrates and products through acclimation of the 1, 3-propylene glycol producing strain, makes the 1, 3-propylene glycol producing strain still grow well under the condition that the substrates with high concentration and a plurality of products exist, and also improves the capacity of the 1, 3-propylene glycol producing strain of producing 1, 3-propylene glycol. As shown by experimental verification, when the 1, 3-propylene glycol producing strain acclimated by the method is utilized for fermentation production under the same fermentation conditions, the synergistic inhibition function of the substrates of the plurality of products on cell growth is obviously less than that of an unacclimated strain, and the biomass (OD<600nm>) and the yield of the 1, 3-propylene glycol are obviously improved as well. The method has important function in the aspect of breeding the 1, 3-propylene glycol producing strain and has good application prospect.

Description

technical field [0001] The invention relates to a method for acclimating 1,3-propanediol producing bacteria and its application. Background technique [0002] 1,3-propanediol is an important chemical raw material, which can be used as an organic solvent for the preparation of inks, printing and dyeing, coatings, medicines, lubricants and antifreeze agents. A monomer with potential application value in the production of similar compounds. At present, there are many known chemical methods for producing 1,3-propanediol, such as using ethylene oxide as a raw material to synthesize 1,3-propanediol through catalytic hydrogenation and acylation; hydration of acrolein to obtain 3-hydroxypropanediol The aldehyde, 3-hydroxypropanal, is regenerated into 1,3-propanediol by catalytic hydrogenation. However, these methods require high temperature, high pressure and complex precious metal catalysts to realize, which not only require large investment, difficulty in separation and purifica...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/18C12R1/22C12R1/01C12R1/225
Inventor 杨光曹竹安裴海生张刚
Owner TSINGHUA UNIV