Preparation method of 2,6-difluorobenzamide by utilizing rhodococcus ruber

A technology of difluorobenzamide and Rhodococcus erythrococcus, applied in the biological field, can solve problems such as high requirements for production equipment and production conditions, low conversion rate and selectivity of the method, and harsh reaction conditions of the production process, so as to reduce production costs, The effect of high substrate tolerance and fast reaction speed

Active Publication Date: 2014-01-15
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0005] In summary, the current production of 2,6-difluorobenzamide has the following deficiencies: 1. Part of the production process has harsh reaction conditions, which require high production equipment and production

Method used

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  • Preparation method of 2,6-difluorobenzamide by utilizing rhodococcus ruber
  • Preparation method of 2,6-difluorobenzamide by utilizing rhodococcus ruber

Examples

Experimental program
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Example Embodiment

[0028] Example 1:

[0029] A method for preparing 2,6-difluorobenzamide by using Rhodococcus rubrum, the preparation steps are:

[0030] ⑴Seed and fermentation culture: Inoculate the Rhodococcus ruber (CGMCC3090) strain into the seed culture medium, culture at 180r / min, 30℃, 28h to obtain the seed culture solution, and transfer it to the fresh fermentation at an inoculum of 3wt% Culture medium, 180r / min, 28°C, culture for 48h to obtain the bacterial fermentation broth:

[0031] Seed medium: glycerol 10g / L, peptone 5g / L, malt powder 3g / L, yeast powder 3g / L, NaCl1g / L, pH7.0;

[0032] Fermentation medium: glucose 20g / L, yeast powder 5g / L, MgSO 4 ·7H 2 O, KH 2 PO 4 , K 2 HPO 4 0.5g / L, urea 7g / L, NaCl 2g / L, CoCl 2 0.02g / L, sodium glutamate 1g / L, pH7.0. Among them, urea and inducer CoCl2 were sterilized separately for 3-5 minutes.

[0033] ⑵ Preparation of Rhodococcus rubrum suspension: centrifuge the above-mentioned bacterial cell fermentation broth, remove the supernatant, and use pH 7.5 ...

Example Embodiment

[0039] Example 2:

[0040] Steps (1), (2) and (4) in this embodiment are the same as those in Embodiment 1, and only step (3) is described below.

[0041] (3) Conversion of 2,6-difluorobenzonitrile: The concentration of substrate 2,6-difluorobenzonitrile is 1mol / L, the reaction volume is 10ml, the final concentration of Rhodococcus erythrococcus is 2.37g / L, and the final concentration is 1% methanol, the conversion system is 50mmol / LpH7.5KH 2 PO 4 -NaOH buffer solution, 30°C, 180r / min, 3h after transformation, prepare pure 2,6-difluorobenzamide.

[0042] After testing, the conversion rate of 2,6-difluorobenzonitrile in this step is 100%, and the selectivity to 2,6-difluorobenzamide is 100%.

Example Embodiment

[0043] Example 3:

[0044] Steps (1), (2) and (4) in this embodiment are the same as those in Embodiment 1, and only step (3) is described below.

[0045] ⑶ Conversion of 2,6-difluorobenzonitrile: the concentration of substrate 2,6-difluorobenzonitrile is 2.5mol / L, the reaction volume is 10ml, the final concentration of Rhodococcus rhodochrous cells is 2.37g / L, adding 1% Tween 80, the conversion system is 50mmol / LpH7.5KH 2 PO 4 -NaOH buffer solution, 30℃, 180r / min, after 24h conversion, prepare pure 2,6-difluorobenzamide.

[0046] After testing, the conversion rate of 2,6-difluorobenzonitrile in this step is 100%, and the selectivity to 2,6-difluorobenzamide is 100%.

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Abstract

The invention relates to a preparation method of 2,6-difluorobenzamide by utilizing rhodococcus ruber. The method comprises the following steps: (1) fermentation and culture of seeds; (2) preparation of rhodococcus ruber suspension fluid; (3) conversion of 2,6-difluorobenzonitrile; (4) preparation of finished products. The invention provides a preparation method of 2,6-difluorobenzamide by utilizing rhodococcus ruber to convert 2,6-difluorobenzonitrile into 2,6-difluorobenzamide. The microorganism conversion method has a very high catalytic efficiency, increases the substrate conversion rate, under a material concentration of 3.5 mol/L, the 2,6-difluorobentrinile conversion rate is as high as 100%, the 2,6-difluorobenamide selectivity is 100%, and no by product 2,6- difluorobenzoic acid is generated. The separation and purification technology of target product is simplified, and the method has the advantages of mild reaction conditions, low requirements on equipment, and easy application to production.

Description

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Claims

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Application Information

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Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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