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Method for fixing nitrile hydratase strain by sodium alginate-polyvinyl alcohol

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

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

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specific Embodiment approach 1

[0008] Specific embodiment one: the method for sodium alginate-polyvinyl alcohol immobilization nitrile hydratase bacterial strain in the present embodiment is as follows: one, the sodium alginate that mass concentration is 3.0%-4%, mass concentration is 6%-9% Mix polyvinyl alcohol, nitrile hydratase bacterial suspension with a concentration of 50000cfu / ml to 80000cfu / ml, diatomaceous earth and activated carbon, and then drop the mixed solution into CaCl with a mass concentration of 3.5%-8%. 2 In the saturated boric acid solution, obtain pellets with a diameter of 3 to 4 mm, then solidify the pellets at a temperature of 4°C for 24 hours, and then filter out the pellets; Cross-linking in a glutaraldehyde solution below 2% for 20 minutes to obtain sodium alginate-polyvinyl alcohol immobilized pellets; wherein the concentration of nitrile hydratase bacteria suspension and mass The volume ratio of sodium alginate with a concentration of 3.0%-4% is 1:10-30, the nitrile hydratase ba...

specific Embodiment approach 2

[0012] Embodiment 2: This embodiment differs from Embodiment 1 in that the mass concentration of sodium alginate described in step 1 is 3.5%. 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 1:11-28. 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 polyvinyl alcohol, 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, polyvinyl alcohol, Nitrile hydratase suspension, diatomite and active carbon are uniformly mixed, the mixture is dipped into saturated boric acid solution containing CaCl2; 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 glutaraldehyde solution to be crosslinked for 20 minutes. The relative enzymatic activity of the sodium-alginate-and-polyvinyl-alcohol-fixed pellets produced by the method is increased by 24.8 percent compared with the sodium alginate embedding method, and increased by 30.8 percent compared with the polyvinyl alcohol embedding method. Meanwhile, the fixed Nitrile hydratase obtained by the method for fixing Nitrile hydratase strain by adopting the sodium alginate and the polyvinyl alcohol can carry out seven batches of continuous reactions, thus realizing 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. The advantages of immobilized cells prepared by polyvinyl alcohol embedding method are strong resistance to microbial decomposition, high mechanical stress, and stable chemical properties. Microorganisms immobilized by sodium alginate embedding method have the advantages of less enzyme activity loss, higher mechanical strength, simple preparation, and less toxic...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N11/08
Inventor 秦智张蕾孙晓君
Owner HARBIN UNIV OF SCI & TECH
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