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Method for fixing nitrile hydratase strain by sodium alginate-chitosan microcapsules

A technology of nitrile hydratase bacteria and sodium alginate, which is applied in the direction of fixing on/in the organic carrier, can solve the problems of less reaction batches and large loss of enzyme activity, and achieve the effect of good operational stability

Inactive Publication Date: 2009-02-04
HARBIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
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  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to provide a method for immobilizing nitrile hydratase strains with sodium alginate-polyvinyl alcohol in order to solve the problems of large enzyme activity loss and few reaction batches of the nitrile hydratase strains fixed by the existing method

Method used

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  • Method for fixing nitrile hydratase strain by sodium alginate-chitosan microcapsules
  • Method for fixing nitrile hydratase strain by sodium alginate-chitosan microcapsules
  • Method for fixing nitrile hydratase strain by sodium alginate-chitosan microcapsules

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specific Embodiment approach 1

[0008] Specific embodiment one: the method for sodium alginate-chitosan microcapsule immobilization nitrile hydratase bacterial strain in the present embodiment is as follows: one, the sodium alginate that mass concentration is 3.0%-4%, concentration is 50000cfu / ml~80000cfu The nitrile hydratase bacterial suspension, diatomaceous earth and activated carbon of / ml are mixed evenly, then the mixed solution is dripped into the CaCl with a mass concentration of 3.5%-8%. 2 In the solution, after obtaining pellets with a diameter of 2.5-3.5mm, solidify the pellets at a temperature of 4°C for 24 hours, and then filter out the pellets; 2% and chitosan mass fraction of 1.2%-1.8% chitosan acetic acid solution for 20min-40min, then filter out the pellets, and then wash 3 times with distilled water to obtain sodium alginate-chitosan Microcapsules; wherein the volume ratio of the nitrile hydratase bacterial suspension with a concentration of 50000cfu / ml to 80000cfu / ml and the sodium algina...

specific Embodiment approach 2

[0012] Specific embodiment two: the difference between this embodiment and specific embodiment one is that the concentration in step one is the volume of the nitrile hydratase bacterial suspension of 50000cfu / ml~80000cfu / ml and the mass concentration of 3.0%-4% sodium alginate The ratio is 1:15-25. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0013] Specific embodiment three: the difference between this embodiment and specific embodiment one is that the concentration in step one is the volume of the nitrile hydratase bacterial suspension with a concentration of 50000cfu / ml~80000cfu / ml and the sodium alginate with a mass concentration of 3.0%-4%. The ratio is 1:20. Others are the same as in the first embodiment.

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Abstract

The present invention discloses a method for fixing Nitrile hydratase strain by adopting sodium alginate and chitosan microcapsules, which relates to a method for fixing Nitrile hydratase strain. The present invention resolves the problems that the loss of the enzymatic activity of the Nitrile hydratase strain fixed by the prior method is large and that the number of reaction batches is little. The method includes the following steps: after sodium alginate, Nitrile hydratase suspension, diatomite and active carbon are uniformly mixed, the mixture is dipped into CaCl2 solution; the produced pellets are solidified under the temperature of 4 DEG C for 24 hours; and the pellets are filtered out and then put into chitosan acetic acid solution to be filmed. The sodium alginate and the chitosan microcapsules, which are obtained by the method, have the highest relative enzymatic activity, reaching 91.6 percent, increased by 23.3 percent compared with the relative enzymatic activity of the sodium alginate and the chitosan microcapsules obtained by the sodium alginate embedding method. The Nitrile hydratase strain obtained by the method can realize ten batches of reactions, and compared with the sodium alginate embedding method, the method has better operation stability and continuity and can realize industrialized continuous production.

Description

technical field [0001] The invention relates to a method for immobilizing nitrile hydratase bacterial strains. Background technique [0002] Nitrile hydratase (Nitrile hydratase, Nhase) enzymatic classification number is EC4.2.1.84, is a class of enzymes that can catalyze the conversion of nitriles into corresponding amides. The main industrial application is to catalyze the hydration of acrylonitrile to produce acrylamide . At present, the immobilization of nitrile hydratase mainly adopts polyacrylamide gel embedding and cutting method in Japan, and sodium alginate embedding method is generally used in my country. Microorganisms immobilized by sodium alginate embedding method have the advantages of less enzyme activity loss, higher mechanical strength, simple preparation, and less toxicity to organisms, but they are unstable in solutions containing polyvalent anions and high-concentration electrolytes, especially in phosphoric acid. In the salt system, calcium ions are ea...

Claims

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Application Information

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IPC IPC(8): C12N11/10
Inventor 秦智张蕾孙晓君
Owner HARBIN UNIV OF SCI & TECH
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