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Culture medium and method for producing laccase by Ascheronia placenta fermentation

A technology of Ascospora and culture medium, which is applied in the field of medium for fermenting and producing laccase from Ascosporidium, achieving the effects of high laccase yield, easy control and mild conditions

Inactive Publication Date: 2013-03-27
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The fungus belongs to Ascomycota (Ascomycota), Faecalimycetes (Sordariomycetes), Hypocreales (Hypocreales), Ergotaceae ( Clavicipitaceae) Ascheronia genus (Aschersonia), but there are few domestic reports on the production of laccase from Ascheronia platina, which has great development prospects

Method used

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  • Culture medium and method for producing laccase by Ascheronia placenta fermentation
  • Culture medium and method for producing laccase by Ascheronia placenta fermentation
  • Culture medium and method for producing laccase by Ascheronia placenta fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Single factor experiment to determine the optimal composition of culture medium

[0023] (1) Preparation of seed medium and basal medium

[0024] (a) Seed medium: 1L of 20% potato juice, 2% glucose, 1% yeast extract, natural pH. 1.01MPa, sterilize for 20min.

[0025] (b) Basal medium: 1L of 20% potato juice, 2% glucose, 1% yeast extract, natural pH. 1.01MPa, sterilize for 20min.

[0026] (2) Production of fermented seeds: inoculate Sophora platinospores on potato agar medium, culture at 25°C for 5 days, and activate after spore production; use the inoculation needle to inoculate the spore powder to the seeds of the activated strain The culture medium was cultured at 25°C for 24 hours at a rotational speed of 160 r / min, which was the fermentation broth.

[0027] (3) Preparation of enzyme liquid: take the fermentation liquid in step (2) and centrifuge at 5000 r / min for 15 min at 4°C, and the supernatant is the crude enzyme liquid for use. The absorbance was measured ...

Embodiment 2

[0045] Orthogonal experiment to determine optimal fermentation conditions

[0046] (1) Orthogonal experiment to determine the best medium

[0047] On the basis of the culture medium determined by the single factor experiment, the solubility and pH value of each component were changed, and different culture media were configured, and the method was the same as that of steps (1) and (2) in Example 1. See Table 1. Inoculate 250mL of the same amount of inoculum (10%) into 250mL of culture solution, and culture at 25°C with a rotation speed of 160 r / min for 3 days. Then measure the enzyme activity according to the steps (3) and (4) of Example 1. Select the experimental group number with the optimal result of the orthogonal experiment and the highest enzyme production, and carry out the verification experiment. In order to avoid being cumbersome, the preparation of the culture medium, the preparation of the crude enzyme solution, and the enzyme activity assay method in the embodim...

Embodiment 3

[0065] Fungal Laccase Fermentation Method

[0066] (1) Preparation of fermented seeds

[0067] (A) Strain and culture medium

[0068] Bacteria: Cyclospora

[0069] Slant medium: PDA medium

[0070] Liquid seed medium: 1L of 20% potato juice, 2% glucose, 1% yeast extract, natural pH.

[0071] (B) Preparation of seed solution

[0072] Transfer the pure Septoria platina on the slant to several 250mL Erlenmeyer flasks filled with 100mL of culture medium, and then cultivate them at 25°C for 3 days on a reciprocating shaker at a speed of 160 r / min. When the silk grows vigorously and the bacterial liquid is viscous, it means that the seeds have grown well.

[0073] (2) Fermentation culture

[0074] Fermentation medium: According to the results of the orthogonal experiment of medium optimization, that is, containing 0.5w% glucose, (NH 4 ) 2 SO 4 4w%, Zn 2+ 0.5×10 -4 mol / L, V B4 5×10 -4 w %, the initial pH of the medium is 4.4, and it is prepared.

[0075] With the Se...

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Abstract

The invention relates to a fungal fermentation culture medium and process. More particularly, the invention relates to a culture medium and method for producing laccase by Ascheronia placenta fermentation. The raw materials of the culture medium comprise 1-4wt% of glucose, 1-4wt% of (NH4)2SO4, 0.5*10<-4>-4*10<-4> mol / L of Zn<2+>, and 1*10<-4>-1*10<-3> wt% of VB4; and the initial pH of the culture medium is 3.8-4.4. According to the method, a fermented seed solution is inoculated to the culture medium, the liquid containing quantity of a 250 mL triangular flask is 50 mL, the inoculum size is 1mL, the number of glass beads is zero, and laccase is cultured at 25 DEG C at the rotational speed of 160 r / min. The fermentation method is short in fermentation period, mild in condition, and easy to control; and the culture medium provided for producing laccase by Ascheronia placenta fermentation has the advantages that the laccase yield is high, the laccase activity is high, and the like.

Description

technical field [0001] The invention relates to a fungal fermentation medium and a process. More specifically, the present invention relates to a culture medium and method for fermenting and producing laccase from S. platinospores. Background technique [0002] Laccase is a copper-containing polyphenol oxidase that catalyzes the one-electron oxidation of a range of organic and inorganic compounds, accompanied by the aerobic reduction to water. It has been found that a variety of organisms can produce laccase, including plants, fungi, insects, bacteria and so on. Among them, white rot fungi are the most widely distributed (Chao Yapeng et al., 2001). [0003] Laccase has a wide range of substrates and can catalyze the oxidation of a variety of phenols and aromatic amines to degrade polycyclic aromatic hydrocarbons. Therefore, laccase has important application value in biodegradation, pulping and papermaking, wastewater treatment, biobleaching, detoxification of pollutants, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12R1/645
Inventor 邱君志苏礼超吴宝杰李小霞涂洁宋飞飞邱云锋郭庆丰何肖云毛丽慧
Owner FUJIAN AGRI & FORESTRY UNIV
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