Method for directly immobilizing tannase in fermentation liquor

A technology of immobilizing enzymes and tannases, which is applied in the direction of immobilizing on or in inorganic carriers, can solve the problems of restricting the large-scale application of tannases, the high cost of tannases, and the high cost of tannases. Achieve good reusability and magnetic manipulation performance

Active Publication Date: 2015-10-07
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The purification of tannase is a complex and expensive process, which is also the key reason for

Method used

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  • Method for directly immobilizing tannase in fermentation liquor
  • Method for directly immobilizing tannase in fermentation liquor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Preparation of immobilized tannase by direct immobilization of magnetic nanoparticles

[0028] (1) Will Aspergillus tubingensis CICC 2651 slant strains were transferred to PDA slant and activated for 96 h, and the slant spores were washed with 0.8% sterile Tween-80 solution to prepare the concentration of 1.5×10 7 spore suspension per mL, inoculate 5 mL of spore suspension in a 250 mL shake flask (the medium is PD, that is, PDA-removed agar, the filling volume is 100 mL, and 5 glass beads with a diameter of 0.5 cm are added to each bottle) 28 Cultivate for 72 h at ℃ to obtain the fermented seed liquid of Aspergillus tubingensis.

[0029] (2) Take the dried Tieguanyin tea stalk powder and pass it through a 40-mesh sieve, add 9% sucrose, 3% glucose, 6% ammonium sulfate and 0.5% yeast extract, and the solid-water ratio is 1:2 (m:v) , sterilized at 121°C for 15 min and cooled to room temperature to obtain the fermentation medium. Each gram of fermentation me...

Embodiment 2

[0033] Example 2: Preparation of immobilized tannase by glutaraldehyde cross-linking method of magnetic nanoparticles

[0034] (1) Will Aspergillus tubingensis CICC 2651 slant strains were transferred to PDA slant and activated for 96 h, and the slant spores were washed with 0.8% sterile Tween-80 solution to prepare the concentration of 1.5×10 7 spore suspension per mL, inoculate 5 mL of spore suspension in a 250 mL shake flask (the medium is PD, that is, PDA-removed agar, the filling volume is 100 mL, and 5 glass beads with a diameter of 0.5 cm are added to each bottle) 28 Cultivate for 72 h at ℃ to obtain the fermented seed liquid of Aspergillus tubingensis.

[0035] (2) Take the dried Tieguanyin tea stalk powder and pass it through a 40-mesh sieve, add 9% sucrose, 3% glucose, 6% ammonium sulfate and 0.5% yeast extract, and the solid-water ratio is 1:2 (m:v) , sterilized at 121°C for 15 min and cooled to room temperature to obtain the fermentation medium. Each gram of f...

Embodiment 3

[0039] Example 3: Preparation of immobilized tannase by covalent binding of magnetic nanoparticles carbodiimide

[0040] (1) Will Aspergillus tubingensis CICC 2651 slant strains were transferred to PDA slant and activated for 96 h, and the slant spores were washed with 0.8% sterile Tween-80 solution to prepare the concentration of 1.5×10 7 spore suspension per mL, inoculate 5 mL of spore suspension in a 250 mL shake flask (the medium is PD, that is, PDA-removed agar, the filling volume is 100 mL, and 5 glass beads with a diameter of 0.5 cm are added to each bottle) 28 Cultivate for 72 h at ℃ to obtain the fermented seed liquid of Aspergillus tubingensis.

[0041] (2) Take the dried Tieguanyin tea stalk powder and pass it through a 40-mesh sieve, add 9% sucrose, 3% glucose, 6% ammonium sulfate and 0.5% yeast extract, and the solid-water ratio is 1:2 (m:v) , sterilized at 121°C for 15 min and cooled to room temperature to obtain the fermentation medium. Each gram of ferment...

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Abstract

The invention discloses a method for directly immobilizing tannase in fermentation liquor. The method includes (1), fermenting Tie Guanyin tea stems by the aid of aspergillus tubingensis to obtain free crude tannase; (2), utilizing hydrophilic super-paramagnetic nano-particles as carriers for immobilizing free tannase; (3), immobilizing the free tannase on the nano-particles by a direct immobilizing process, a glutaraldehyde cross-linking process and a carbodiimide covalent binding process so as to obtain liquid immobilized tannase; (4), carrying out vacuum freeze-drying on the liquid immobilized tannase to obtain powder immobilized tannase. The aspergillus tubingensis is used as an enzyme producing strain. The method has the advantages that the magnetic nano immobilized tannase is directly immobilized in the fermentation crude enzyme liquor and accordingly can be easily and quickly prepared; the immobilized tannase is excellent in enzyme activity, storage stability, reusability and magnetic control performance.

Description

Technical field [0001] The present invention involves biological fermentation and enzyme fixed technology fields, especially a method that directly fixed tanninase directly from the fermentation crude enzyme solution. Background technique [0002] Tannase has extremely important application value in food, medicine, feed, especially in the tea beverage industry.At present, tannine enzymes are mainly obtained through microbial fermentation, and a series of purification tannin enzymes are obtained through a series of separate purification operations.The purification of tannine enzyme is a complex and expensive process, which is also a key reason for the high cost of tanninase, and ultimately restricts the large -scale application of tanninase in the industry. [0003] Enzyme fixation technology is a technology that fix free enzymes on a specific carrier by adsorbing, cross -linking, binding of covalent or embedding, which can significantly improve the stability and reuse rate of enz...

Claims

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Application Information

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IPC IPC(8): C12N11/14
Inventor 肖安风吴昌正倪辉蔡慧农杨秋明杜希萍李利君黄高凌陈艳红
Owner JIMEI UNIV
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