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Rhodococcus ruber and method for preparing 5-cyanovaleramide by utilizing same

A technology of cyanovaleramide and Rhodococcus erythrococcus, which is applied in the field of biochemical engineering, can solve the problems of waste of raw materials, harsh reaction conditions, and many by-products, and achieve the effects of simplified separation and purification process, high catalytic efficiency, and short reaction cycle

Active Publication Date: 2010-01-06
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In summary, the current production methods of 5-cyanopentamide have the following deficiencies: 1. The reaction conditions of some methods are harsh, and the requirements for production equipment and production conditions are relatively high, which increases production costs; 2. The conversion rate of some production methods and The selectivity is low, the raw material utilization rate is low, and the waste of raw materials is caused; 3. In the above-mentioned several production methods, there are many by-products produced while generating the target product, which increases the difficulty of product separation and purification

Method used

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  • Rhodococcus ruber and method for preparing 5-cyanovaleramide by utilizing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A kind of method utilizing Rhodococcus rubrum to prepare 5-cyanopentanamide, the preparation steps are:

[0032] (1) Seed culture: add 30mL seed medium to a 250mL shake flask, pick a ring from the preserved slant strains and inoculate it into the seed medium, cultivate at 30°C and 180rpm for 28 hours to obtain a seed solution;

[0033] Seed medium: 1% glycerol, 0.5% peptone, 0.3% malt powder, 0.3% yeast powder, 0.1% NaCl, pH 7.2, sterilized at 121°C for 20 minutes.

[0034] (2) Fermentation culture: Add 30mL fermentation medium into a 250mL shake flask, insert the seed solution according to the inoculum size of 3%, and cultivate at 28°C and 180rpm for 48h;

[0035] Fermentation medium: 2% glucose, 0.5% yeast powder, 0.05% K 2 HPO 4 , 0.05% KH 2 PO 4 , 0.4% MgSO4 ·7H 2 O, 0.7% urea, 0.2% NaCl, 0.002% CoCl 2 , 0.1% sodium glutamate, pH 7.2, sterilized at 115°C for 20min; the inducer CoCl 2 and urea are sterilized separately, CoCl 2 Sterilize at 121°C for 20 minut...

Embodiment 2

[0045] Steps (1), (2), (3) and (5) in this embodiment are all the same as in Embodiment 1, and only step (4) is described below.

[0046] (4) Preparation of the crude product of 5-cyanovaleramide: the substrate adiponitrile concentration is 0.5mol / L, the reaction volume is 30mL, the cell concentration of Rhodococcus erythrococcus is 0.53g / L, 2.5% methanol is added, and the transformation system is 20mmol / L pH7.5 Tris-HCl buffer solution, the temperature is 30°C, the whole reaction is carried out in a water bath reactor, the rotation speed is 180rpm, and the conversion time is 120min. After the transformation, the transformation solution was centrifuged to remove the bacterial cells, and 20% ethanol was added to remove the water by rotary evaporation to obtain the crude product of 5-cyanovaleramide;

[0047] It is detected that the conversion rate of adiponitrile in this step reaches 100%, and the selectivity to 5-cyanovaleramide is 100%.

Embodiment 3

[0049] Steps (1), (2), (3) and (5) in this embodiment are all the same as in Embodiment 1, and only step (4) is described below.

[0050] (4) Preparation of the crude product of 5-cyanovaleramide: the substrate adiponitrile concentration is 0.8mol / L, the reaction volume is 30mL, the cell concentration of Rhodococcus erythrococcus is 0.53g / L, 2.5% methanol is added, and the transformation system is 20mmol / L pH7.5 Tris-HCl buffer solution, the temperature is 30°C, the whole reaction is carried out in a water bath reactor, the rotation speed is 180rpm, and the conversion time is 240min. After the transformation, the transformation solution was centrifuged to remove the bacterial cells, and 20% ethanol was added to remove the water by rotary evaporation to obtain the crude product of 5-cyanovaleramide;

[0051] It is detected that the conversion rate of adiponitrile in this step reaches 100%, and the selectivity to 5-cyanovaleramide is 100%.

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Abstract

The invention relates to a rhodococcus ruber and a method for preparing 5-cyanovaleramide by utilizing the same. The strain needs oxygen, does not need power, spores and gram-positive bacteria, is already stored in the Common Microorganism Center of Chinese Microorganism Strain Storage Management Committee and has storage data of Jun. 4, 2009 and a preservation number of CGMCC No.3090. The method for preparing the 5-cyanovaleramide by utilizing the rhodococcus ruber comprises the following steps: (1) culturing seeds; (2) fermenting and culturing; (3) preparing a rhodococcus ruber suspension; (4) preparing a crude product of 5-cyanovaleramide; and (5) purifying the 5-cyanovaleramide. The invention provides a method for generating the 5-cyanovaleramide by selectively hydrolyzing adiponitrile in a rhodococcus ruber region, has high catalytic efficiency, high adiponitrile conversion rate up to 100 percent and 5-cyanovaleramide regioselectivity up to 100 percent, does not generate a side product of adipamide and simplifies the separation purification process of the 5-cyanovaleramide.

Description

technical field [0001] The invention belongs to the technical field of biochemical engineering, and relates to rhodococcus rubrum and its application, in particular to rhodococcus rubrum and a method for preparing 5-cyanopentamide by using the fungus. Background technique [0002] 5-cyanovaleramide (5-cyanovaleramide, 5-CVAM) can be used to synthesize azafenidin (Azafenidin) and 6-aminocaproamide, wherein azafenidin (Azafenidin) can be used for the weeding of the prepared plant seeds before emergence 6-aminocaproamide is an important intermediate for the synthesis of caprolactam, which is a commonly used organic chemical raw material. 5-cyanopentamide can be prepared by chemically or biocatalyzing the hydration of adiponitrile. Since the molecular formula of adiponitrile contains two cyano groups, it is required that the catalyst can regioselectively hydrolyze one of the cyano groups to generate 5 - cyanovaleramide. [0003] According to retrieval, U.S. Patent No. 5,347,03...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/01
Inventor 王敏李晓丹申雁冰骆健美郑宇孙华
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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