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Laccase preparation method

A laccase and seed technology, applied in the biological field, can solve the problem of low enzyme activity of laccase

Inactive Publication Date: 2015-06-17
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] A variety of laccase-producing Pleurotus fungi have been found, such as Pleurotus pectinosa, Pleurotus erythrosum, Pleurotus rotundum, etc., but the laccase activity is generally not high

Method used

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Embodiment 1

[0028] Slant medium: glucose 20g / L, peptone 10g / L, yeast powder 2g / L, agar 15g / L, and the rest is peanut shell extract; the preparation method of the peanut shell extract is: peanut shell 100g, water 1L, boiling After 45 minutes, filter, and the filtrate is the peanut shell extract.

Embodiment 2

[0030] Slope medium: glucose 5g / L, peptone 20g / L, yeast powder 3g / L, agar 15g / L, and the rest is peanut shell extract; the preparation method of the peanut shell extract is: peanut shell 10g, water 1L, boiling 60min, filter, and the filtrate is the peanut shell extract.

Embodiment 3

[0032] Incline medium: glucose 15g / L, peptone 2.5g / L, yeast powder 5g / L, agar 15g / L, and the rest is peanut shell extract; the preparation method of the peanut shell extract is: peanut shell 100g, water 1L, Boil for 50 minutes, filter, and the filtrate is the peanut shell extract.

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Abstract

The invention discloses a laccase preparation method, which comprises the steps of: (1) inoculating a solid spawn of an inclined medium inoculated with Lentinus tigrinus with a preservation number of CGMCC No.8301 into a liquid seed medium to prepare a seed liquid; (2) inoculating the seed liquid into a fermentation medium, which is in a 500mL triangular flask with a liquid loading volume of 100-250mL, and conducting culture at 25-35DEG C in a shaking table of 150-350rpm for 7-20 days so as to obtain Lentinus tigrinus mycelium fermentation broth; and (3) centrifuging the mycelium fermentation broth at 10000*g for 10min, and taking the supernatant i.e. a laccase crude enzyme fluid. The method provided by the invention utilizes the edible fungus Lentinus tigrinus to conduct liquid submerged fermentation for high yielding of food grade laccase. According to the invention, the laccase in the Lentinus tigrinus mycelium fermentation broth has high enzyme activity, and the enzyme activity of laccase in the fermentation broth is up to 462.59U / mL.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, it relates to a method for producing laccase by submerged fermentation of a tiger skin shiitake mushroom (Lentinus tigrinus). Background technique [0002] Laccase is a copper-containing polyphenol oxidase in the same subfamily as ascorbate oxidase in plants and blue copper oxidases in mammals. According to its source, laccase can be divided into two categories: rhus laccase and fungal laccase. Plant laccase is closely related to the biosynthesis of lignin. Fungal laccase has a variety of physiological and biochemical functions, such as Morphogenesis, fungal-plant host interaction, stress defense, and lignin degradation, etc. According to statistics, there are more than 250 kinds of catalyzed substrates of laccase, mainly including phenols, aromatic amines, carboxylic acids, metal organic compounds, some steroid hormones and biological pigments. Due to the wide range of substrat...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12R1/645
CPCC12N9/0061C12Y110/03002
Inventor 姜文侠孙瑞雪刘琦李晓辉杨萍
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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