Preparation method of bionic immobilized 3-cyanopyridine nitrile hydratase

A technology of cyanopyridine nitrile hydration and nitrile hydratase, which is applied in the field of microorganisms, can solve the problems of many reaction steps, cumbersome preparation of enzyme-producing cells, and large amount of catalyst used in chemical methods, so as to achieve high recovery rate and improve the stability of repeated use Effect

Inactive Publication Date: 2015-09-09
ANHUI COSTAR BIOCHEM CO LTD
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Problems solved by technology

[0006] Chinese patent (CN1424402A) reports a method for the production of nicotinamide by microbial catalysis, which uses the nitrile hydratase of Corynebacterium propionici to convert 3-cyanopyridine into nicotinamide, avoiding the use of a large amount of chemical catalyst and the reaction There are many steps, but the preparation of enzyme-producing cells is relatively cumbersome

Method used

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Embodiment Construction

[0026] A preparation method of biomimetic immobilized nitrile hydratase adopts biomimetic immobilized enzyme, the enzyme is formed by Brevibacterium flavum nitrile hydratase embedded in indium oxide nano shell, the specific preparation method comprises the following steps:

[0027] (1) Preparation of liposome template: Add 15 g of soybean lecithin and 5.5 g of cholesterol into 85 mL of ethanol solution, stir evenly; remove ethanol by rotary evaporation at 40°C until a uniform and transparent film is formed on the reactor; then add 10 mL of 0.5 g / L Brevibacterium flavum nitrile hydratase solution (prepared with 0.1 mol / L, pH=7 disodium hydrogen phosphate-citrate buffer) was used to dissolve the film, and the final concentration was 20 g / L liposome solution;

[0028] (2) Preparation of liposome microcapsules: take the above liposome solution, and slowly drop 1 mL of the inducer trimellitic acid (the concentration of trimellitic acid is 1.5 g / L) under the condition of vigorous s...

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Abstract

The invention discloses an indium oxide nanosphere coating brevibacterium flavum nitrile hydratase, a preparation method and application. The multilayered structure of cells is simulated by referring to the existence form of the hydratase in a living body, and the indium oxide nanosphere is prepared through bionic design of components, functions and processes and used for embedding of the brevibacterium flavum nitrile hydratase. The characteristics that the immobilized nitrile hydratase is high in stability, high in activity, easy to recycle and the like are used for catalyzing 3-cyanopyridine to be transformed into nicotinamide in an efficient hydrolysis mode. The preparation method of the bionic immobilized hydratase can provide reference significance for preparation of a microcyst immobilized carrier, and provide a novel research idea for biotransformation of the 3-cyanopyridine.

Description

technical field [0001] The invention belongs to the technical field of microorganisms. Specifically relates to a preparation method of bionic immobilized 3-cyanopyridine nitrile hydratase. Background technique [0002] Enzymes are a class of highly efficient and specific biocatalysts. However, the high-level structure of the enzyme is very sensitive to the environment. High temperature, high pressure, metal ions and extreme pH environments may inactivate the enzyme. Even if it is not under the optimal reaction conditions, it will gradually become inactivated with the extension of the reaction time. and the enzyme has the disadvantage of being difficult to recycle and reuse, and the existence of the enzyme also brings certain difficulties to the separation of the product, so the application of the enzyme in vitro is severely limited. Immobilization of enzymes can effectively solve the above problems. [0003] The existing immobilization methods include physical adsorption,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02
Inventor 王红伟孔军军强金凤史玉龙
Owner ANHUI COSTAR BIOCHEM CO LTD
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