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Method for synthesizing nicotinamide under catalytic effect of high-molecular-weight nitrile hydratase engineering bacteria

A nitrile hydratase engineering and high molecular weight technology is applied in the field of high molecular weight nitrile hydratase engineering bacteria to catalyze the synthesis of nicotinamide, and can solve the problems of low production efficiency, long growth cycle of Rhodococcus, and low concentration of final product, and achieve the growth cycle the effect of shortening

Active Publication Date: 2017-05-31
JIANGNAN UNIV
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Problems solved by technology

[0003] At present, in industrial production, Rhodococcus rhodochrous J1 is mainly used to catalyze the production of nicotinamide, and the method of substrate feeding is adopted. However, the growth cycle of Rhodococcus is long, requiring 100h, and the production efficiency is not high, only 162g / (L·h)
[0004] At present, there are also reports on the production of nicotinamide by recombinant bacteria, but the concentration of the final product is low, only 240g / L

Method used

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  • Method for synthesizing nicotinamide under catalytic effect of high-molecular-weight nitrile hydratase engineering bacteria
  • Method for synthesizing nicotinamide under catalytic effect of high-molecular-weight nitrile hydratase engineering bacteria

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Embodiment 1

[0036] (1) High-density fermentation of recombinant genetically engineered bacteria

[0037] Seed culture: Take out the preserved glycerol tube from the -80°C refrigerator, inoculate 200 μL of bacterial liquid into 30 mL of LB medium containing 50 μg / mL kanapenicillin resistance, and place it in a constant temperature shaker at 37°C at 200 r / min for cultivation 7.5h.

[0038] Fermentation tank culture: 5L Winpact automatic fermentation tank is used, and the initial liquid volume is 2L. Set the initial pH to 7, the culture temperature to 37°C, the initial rotation speed to 200r / min, and the ventilation rate to 5L / min. Put the seed solution obtained in the previous step into the fermenter with an inoculum size of 6%, and add kanapenicillin at the same time to a final concentration of 50 μg / mL. The dissolved oxygen in the tank is maintained at 30%, and the dissolved oxygen rebound occurs 5-6 hours after inoculation, at this time, the specific growth rate μ = 0.2h -1 Exponentia...

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Abstract

The invention discloses a method for synthesizing nicotinamide under catalytic effect of high-molecular-weight nitrile hydratase engineering bacteria, and belongs to the field of fermentation engineering. Through building a escherichia coli high-density fermentation method, the bacteria growth period is greatly shortened to 48h from original 100h of original bacteria, so that the time is reduced by 52 percent; on the basis of an existing catalytic process, the concentration of a product obtained through catalyzing is improved by 110.8 percent to achieve 506g / L compared with existing recombinant bacteria; the production efficiency is improved by 212 percent to achieve 508g / (L.h) compared with original rhodococcus.

Description

technical field [0001] The invention relates to a method for catalyzing and synthesizing nicotinamide by high-molecular-weight nitrile hydratase engineering bacteria, and belongs to the field of fermentation engineering. Background technique [0002] Nitrile hydratase (NHase, EC 4.2.1.84) is an enzyme that catalyzes the conversion of nitriles to amides and is widely used in industrial production. Nicotinamide is an important intermediate in the synthesis of drugs and pesticides, and is also widely used in the food industry. [0003] At present, in industrial production, Rhodococcus rhodochrous J1 is mainly used to catalyze the production of nicotinamide, and the substrate feeding method is adopted. However, the growth cycle of Rhodococcus is long, requiring 100 hours, and the production efficiency is not high, only 162g / (L·h). [0004] At present, there are also reports on the production of nicotinamide by recombinant bacteria, but the concentration of the final product i...

Claims

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Application Information

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IPC IPC(8): C12P17/12C12R1/19
CPCC12P17/12
Inventor 周哲敏王哲刘中美郭军玲汪启
Owner JIANGNAN UNIV
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