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Method for preparing laccase by liquid fermentation of Pleurotus ferulae

A technology of liquid fermentation and Ferulicum ferulae, applied in the field of microbial fermentation, can solve the problem of never producing laccase

Active Publication Date: 2013-06-12
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] At present, a variety of Pleurotus fungi with the ability to produce laccase have been found, such as Pleurotus pectinosa (Pleurotus ostreatus), Pleurotus eryngii (Pleurotus eryngii), Pleurotus ring handle (Pleurotus eryngii), etc. Related studies and reports on the production of laccase by fermentation of Pleurotus ferulae (A. ferulae)

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  • Method for preparing laccase by liquid fermentation of Pleurotus ferulae

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Embodiment 1, the tissue separation of ferulae ferulae bacterial classification

[0018] The fresh unopened fruiting bodies of wild ferulae ferulae collected from the Altay region of Xinjiang, the base of the stipe and the outer skin of the stipe were cut off, and the surface of the fruiting body was disinfected with 75% ethanol, and then moved into the ultra-clean workbench for 20 hours of ultraviolet light irradiation. minute. Divide the fruiting body in two with a sterilized and cooled scalpel, cut the epidermal tissue in the stipe or in the stipe, at the junction of the stipe and cap, at the cap and gills, and cut the tissue into soybeans Small pieces of small size, the epidermal tissue of the fruiting body cannot be cut through when cutting. Use sterile tweezers to insert the test tube slant medium (i.e. PDA medium), inoculate 10 test tube slants for each tissue part, culture at 25°C, observe the mycelial growth and contamination every day, and when it grows to 7 ...

Embodiment 2

[0019] Embodiment 2, ferulae ferulae liquid fermentation produces laccase

[0020] (1) Slant culture: inoculate the Ferula ferulae strain obtained in Example 1 on the slant medium (ie PDA medium), and cultivate it at 25°C for about 7 days until the mycelium covers the slant;

[0021] (2) Seed culture: Inoculate the slant bacteria into a 250 ml Erlenmeyer flask containing 60 ml of seed medium. The composition of the seed medium (unit g / L): glucose 20, corn flour 20, bran 10, KH 2 PO 4 3. MgSO 4 ·7H 2 o 2. The pH is natural, sterilized at 121°C for 20 minutes; cultivated at 25°C for 7 days on a shaker with a rotation speed of 150 rpm;

[0022] (3) Liquid fermentation culture: Take 6 ml of cultured seeds and inoculate them into a 500 ml Erlenmeyer flask containing 150 ml of fermentation medium. Fermentation medium composition (unit g / L): glucose 15, corn flour 10, bran 10, KH 2 PO 4 3. MgSO 4 ·7H 2 O 2, the initial pH is 7.0, sterilized at 121°C for 20 minutes; culti...

Embodiment 3

[0030] Embodiment 3, ferulae ferulae liquid fermentation produces laccase

[0031] Inoculate the cultivated seeds into the fermentation medium, the composition of the fermentation medium (unit g / L): glucose 20, corn flour 20, bran 15, KH 2 PO 4 3. MgSO 4 ·7H 2 O 2 , the initial pH is 9.0, cultured at 26° C. for 7 days, and the other conditions are the same as in Example 2. At the end of the fermentation, the activity of laccase in the fermentation broth reached 8629.5 U / L.

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Abstract

The invention discloses a method for preparing laccase by liquid fermentation of Pleurotus ferulae, belonging to the technical field of microbial fermentation. The method provided by the invention takes Pleurotus ferulae as a production strain, generates laccase by liquid fermentation, and is realized by the following main steps: (1) slant culture; (2) seed culture; and (3) liquid fermentation culture. Cultivated seed culture is inoculated into a liquid fermentation medium at an inoculum size of 4%, and cultivated at 22-26 DEG C for 7 days under the shaker rotation speed of 150rpm, wherein the fermentation medium comprises the following components: 15-20g / L of glucose, 10-20g / L of corn flour, 10-15g / L of wheat bran, 3g / L of KH2PO4 and 2g / L of MgSO4.7H2O, and the initial pH value is 7.0-9.0. The method realizes a goal of generating laccase with Pleurotus ferulae, and the fermentation level of Pleurotus ferulae reaches or exceeds the fermentation level of other Pleurotus fungi. The new method for generating laccase with the microbial fermentation is provided, and in addition, a new direction for application of Pleurotus ferulae in fermentation industry is developed.

Description

technical field [0001] The invention discloses a method for producing laccase by liquid fermentation of Ferula ferulae, which belongs to the field of microbial fermentation. Background technique [0002] Laccase (benzonediol:oxygen oxidoreductase, EC 1.10.3.2), a copper-containing polyphenol oxidase, is widely distributed in plants, bacteria and fungi. In addition, laccase has also been found in some insects. Laccase is a one-electron oxidoreductase, and its catalytic substrates include phenol and its derivatives, arylamine and its derivatives, carboxylic acid and its derivatives, and some metal ions. Due to its wide range of substrates, laccase has great value in the fields of food industry, pulp and paper making, decolorization of artificial dyes, and development of new environmentally friendly materials. [0003] In the food industry, laccases are widely used to control the clarification and color of beverages. The traditional method is to adsorb the beverage through su...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12R1/645
Inventor 丁重阳陈友枝彭林顾正华张梁石贵阳章克昌
Owner JIANGNAN UNIV