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A kind of culture medium and method for producing laccase by fermentation of leather ear fungus

A culture medium formula and culture medium technology are applied in the field of laccase production by fungal culture, and can solve the problems of low laccase yield, long culture period, easy to be polluted, etc. high vitality effect

Inactive Publication Date: 2015-10-21
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, although people have a better understanding of the mechanism of fungal enzyme production, compared with actual needs, most test strains have low synthetic laccase production when grown on artificial and natural media.
Moreover, the culture period of fungi is long, and they are easily polluted. Some require the induction of toxic aromatic compounds, and the enzyme production activity is unstable and relatively low, which makes it difficult to realize the industrial production of laccase.

Method used

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  • A kind of culture medium and method for producing laccase by fermentation of leather ear fungus
  • A kind of culture medium and method for producing laccase by fermentation of leather ear fungus
  • A kind of culture medium and method for producing laccase by fermentation of leather ear fungus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 single factor experiment determines optimum medium composition

[0026] (1) Preparation of seed medium and basal medium

[0027] (a) Seed medium: based on the weight of 1L medium, it contains 15% potato juice, 1% glucose, 0.2% ammonium tartrate, and the pH is natural. 1.01MPa, high pressure steam sterilization for 20min.

[0028] (b) Basal medium: based on the weight of 1L medium, containing 15% potato juice, 1% glucose, 0.2% ammonium tartrate, and natural pH. 1.01MPa, high pressure steam sterilization for 20min.

[0029] (c) Potato agar medium: PDA basal medium

[0030] (2) Production of fermented seeds: inoculate the Auricularia fungus FG-35 (Panus rudis FG-35, preservation number: CCTCC NO: M2013106) on potato agar medium, culture at 31°C for 5 days, and wait for it to produce spores , that is, activation; inoculate the activated spores of the strain into the seed medium with an inoculation needle, and cultivate for 96 hours at 31°C with a rotation s...

Embodiment 2

[0047] Example 2 Response Surface Determination of Optimal Fermentation Medium

[0048] (1) Experiment to determine the best medium

[0049] On the basis of determining the single-factor experimental medium, determine the best components, change the concentration of each component, and configure different mediums, the method is the same as the steps (1) and (2) in Example 1. See Table 1, inoculate the same amount of inoculum (0.5%) seed solution into a 300mL conical flask filled with 50-90mL medium, and the number of spores of the strains inserted is 2×10 6 -5×10 6 About one, cultivated at 28-32°C for 4-8d at a rotational speed of 160-180r / min. Then measure the enzyme activity according to the steps (3) and (4) of Example 1. Three groups of experiments, 1, 5, and 14 with the best results of Plackett-Burman and the steepest climbing experiment and the highest enzyme production, were selected for verification experiments.

[0050] (2) Plackett-Burman screening factors affect...

Embodiment 3

[0074] Embodiment 3 leather ear fungus produces laccase fermentation method

[0075] (1) Preparation of seeds

[0076] (A) Strain and culture medium

[0077] Strain: Auricularia FG-35

[0078] Slant medium: PDA basal medium

[0079] Ingredients of seed liquid medium: 15% potato juice (1L), 1% glucose, 0.2% ammonium tartrate, natural pH.

[0080] (B) Preparation of seed solution

[0081] Transfer the bacteria on the inclined surface into a 300mL Erlenmeyer flask with 50mL of medium, and culture it in a constant temperature shaking incubator at 31°C and 160r / min for 96h, so that the bacterial liquid becomes viscous, indicating that the seeds have grown well up.

[0082] (2) Fermentation culture

[0083] Fermentation medium: the optimal medium optimized according to the response surface: containing soluble starch 10.04g / L, peptone 0.2g / L, K 2 HPO 4 0.50g / L, KH 2 PO 4 0.50g / L, ZnSO 4 ·7H 2 O 0.01g / L, MgSO 4 ·7H 2 O 0.5g / L, CuSO 4 ·7H 2 O 0.01g / L, FeSO 4 ·7H 2 O...

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Abstract

The invention discloses a culture medium for fermenting panus rudis to produce laccase and a laccase production method. A culture medium formula comprises 10.04g / L of soluble starch, 0.2g / L of peptone, 0.50g / L of K2HPO4, 0.50g / L of KH2PO4, 0.01g / L of ZnSO4.7H2O, 0.5g / L of MgSO4.7H2O, 0.01g / L of CuSO4.7H2O, 0.005g / L of FeSO4.7H2O, 0.05g / L of MnSO4, 0.08g / L of CaCl2, 0.1g / L of VB1, 0.4% of tween-40, and 0.05g / L of veratryl alcohol; the initial pH value of the culture medium is 6.8. A fermentation seed solution is inoculated in the culture medium so as to culture and produce the laccase at the temperature of 31 DEG C and the rotating speed of 160r / min. The culture medium for fermenting the panus rudis to produce the laccase, provided by the invention, has the advantages of high laccase productivity, high laccase activity and the like; the culture method is short in period and mild in conditions.

Description

technical field [0001] The invention belongs to the technical field of fungal culture to produce laccase, and in particular relates to a laccase-producing fungal fermentation medium and a process. More specifically, the present invention relates to a culture medium and method for producing laccase by germ fermentation. Background technique [0002] Laccase is a copper-containing polyphenol oxidase that can oxidize a variety of phenols and aromatic amines and simultaneously reduce oxygen molecules to water. Laccase can directly act on lignin and oxidatively degrade phenolic lignin structural units. In the presence of some small molecule mediators capable of transferring electrons, it can oxidatively degrade non-phenolic lignin structural units. Therefore, laccase has shown great research value and application potential in pulp biological bleaching, industrial wastewater treatment, organic dye decolorization and polymer catalytic synthesis. Laccase is considered to be the mos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12R1/645
CPCC12N9/0061C12Y110/03002
Inventor 周国英刘剑杨菁刘君昂李河左杰田媛媛李冬琴刘慧娟
Owner CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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