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Trametes gallica, culture method and application thereof

A technology of Trametes cruzi and a culturing method, applied in the field of microorganisms, can solve the problems of low yield, dependence on metal copper ions, rare fungal strains and the like

Inactive Publication Date: 2008-10-08
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the heat-resistant laccases discovered so far are concentrated in bacteria, but they have limitations such as low yield and dependence on metal copper ions, and high-yield fungal strains are rare

Method used

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  • Trametes gallica, culture method and application thereof
  • Trametes gallica, culture method and application thereof
  • Trametes gallica, culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Isolation and screening of high heat-resistant laccase strains:

[0028] 1. Medium:

[0029] Separation and purification medium: add 200g bran into 1L water, boil for 30min, filter with 6 layers of gauze; weigh 20g agar strips; put the filtered bran liquid and weighed agar strips into a 1000ml Erlenmeyer flask, add water to volume to 1000ml; sterilize the bran juice culture medium and the washed petri dish at 121°C for 20 minutes, add 200mg / L penicillin and 200mg / l streptomycin, and pour it into a plate for later use.

[0030] Laccase selective medium: Add 100g bran to 1L water, boil for 30min, filter with 6 layers of gauze; weigh 20g agar strips; put the filtered bran liquid and weighed agar strips into a 1000ml Erlenmeyer flask, Add water to make the volume to 1000ml; sterilize the bran juice culture medium and the washed petri dish at 121°C for 20 minutes, add 200mg / L guaiacol, and pour it into a plate for later use.

[0031] 2. Screening steps:

[0032] (1) Take ...

Embodiment 2

[0041] Identification of high thermostable laccase producing strains:

[0042] 1. Morphological and physiological and biochemical identification

[0043] Firstly, the collected fruiting body or the fruiting body produced by culture and spore morphology were compared with the fungal taxonomy manual "Fungi Identification Manual" (written by Wei Jingchao) to preliminarily identify that the strain's fruiting body is sessile, lateral, and suberin. Cap 1.5-5.0cm×2-8cm, 5-25mm thick, with white or yellow-white coarse hair bundles and concentric rings. The flesh of the fungus is white to yellowish white, basidiospore club-shaped, about 10μm×3μm, very similar to the characteristics of the genus Trametes, and should belong to the genus Trametes. The specific species name needs to be identified by molecular biology methods.

[0044] 2. Sequence Analysis of ITS

[0045] (1) Medium for sequence analysis:

[0046] Mycelia culture medium: mix 10% bran with 90% deionized water by weight, b...

Embodiment 3

[0084] Preparation of heat-resistant laccase and determination of heat resistance

[0085] Firstly, the strains stored on the slant of the bran extract liquid stored in the refrigerator at 4°C were transferred and activated, then transferred to fresh PDA solid slant medium or bran extract liquid medium for 5 days, and then the transfer expansion culture was carried out. The inoculum size depends on the volume of the expanded culture. Enzyme production medium is bran 200±5g added to 1L water, boiled for 30 minutes, filtered through six layers of gauze, natural pH, sterilized at 121°C for 20 minutes. The culture time is 6-8 days, the culture temperature is 28-30° C., and the shaking condition is an oscillating shaker at 70 rpm or other shakers with similar conditions. The culture is suction-filtered with a G4 sand core funnel or centrifuged to remove the mycelium to obtain a crude enzyme liquid, and a purified laccase can be obtained by conventional purification methods.

[00...

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Abstract

The invention discloses bacterial strain trametes hirsuta 1g-9 produced by heat resistance laccase and a culturing method and the application thereof, which belongs to the technical field of the microorganism. The preservation number of the bacterial strain trametes hirsute 1g-9 is CGMCC No.2422; the bacterial strain trametes hirsute 1g-9 can be cultured through mill offal leach liquor slant culture or activated through PDA solid slant culture medium, to ferment the culture medium through the mill offal leach liquor to enlarge the culture; the bacterial strain trametes hirsuta 1g-9 can be applied to the industrial papermaking, the paper pulp modification, the papermaking sewage treatment, the dyeing and printing sewage treatment, the food industry, the beverage industry, the clothing industry and the organic synthesis industry.

Description

(1) Technical field [0001] The invention relates to a basidiomycete, in particular to a heat-resistant laccase-producing fungal strain Trametes tenaci, which belongs to the technical field of microorganisms. (2) Background technology [0002] Laccase is a metal ion-containing glycoprotein belonging to the multi-copper oxidase family, which is distributed in animals, plants, fungi, and bacteria, especially in fungi and bacteria. Laccase can use oxygen as an electron donor to directly oxidize aromatic compound substrates or mediators, and can also oxidize macromolecular lignin or non-phenolic aromatic compounds with high redox potential under the mediation of mediators. [0003] Thermostability is a bottleneck that limits the wide application of laccase, and the isolation of strains with good thermostability is of great significance for expanding the application range of laccase, prolonging the application time of laccase, and reducing the application cost of laccase. Laccase...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/04C02F3/34A23L1/03A23L2/84A23L3/3571D21C9/00D21C3/00C12S13/00C12R1/645A23L29/00C12S99/00
Inventor 黄峰张海波张应龙
Owner SHANDONG UNIV
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