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Laccase, preparing method thereof and strain special for same

A strain and laccase technology, applied in the field of producing laccase, methods and special strains thereof, can solve the problem of low laccase enzyme activity, and achieve the effects of high laccase enzyme activity, simplified extraction process and easy extraction

Inactive Publication Date: 2007-04-04
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The laccase activity of the known domestic strains is relatively low, up to 21U / ml

Method used

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  • Laccase, preparing method thereof and strain special for same
  • Laccase, preparing method thereof and strain special for same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1, the isolation of Microporosa hemorrhoids mk528

[0020] P. haemoporus mk528 was isolated from the dikaryotic hyphae of P. haemopore G5, and G5 was obtained from the dikaryotic hyphae obtained from the fruiting body of P. haemorrhoids (Tang Shun, Lomascolo A, Guo Lin, etc. (2001) Isolation of white-rot fungi from China for screening laccase-hyperproducing strains. Fungi System 20:520-525).

[0021] Microporosa haemophilus G5 on agar containing 20g / l -1 and malt extract 20g / l -1 After culturing for 7 days, the culture dish was inverted and kept at room temperature for 3-4 weeks. Then use high-temperature sterilized distilled water to collect basidiospores from the lid of the petri dish, then dilute these basidiospore suspensions by an appropriate multiple, and spread them to contain agar 20g / l -1 and malt extract 20g / l -1 culture dish at room temperature, and on the second or third day, the visible basidiospore colonies were separated from the single spo...

Embodiment 2

[0022] Embodiment 2, fermenting and cultivating Microporosa haemorrhoids mk528 produces laccase

[0023] 1. Fermentation of Microporosa haemophilus mk528

[0024] Solid seed medium: 2% malt extract, 2% agar.

[0025] Fermentation medium: maltose 15g, ammonium tartrate 2g, KH 2 PO 4 1.333g, NaH 2 PO 4 12H 2 O 0.39g, MgSO 4 ·7H 2 O 0.5g, sodium succinate (CH 2 COONa) 2 ·6H 2 O 1.18g, FeSO 4 ·7H 2 O 0.0315g, CaCl 2 2H 2 O0.1g, MnSO 4 ·H 2 O 0.035g, CH 3 COONa·3H 2 O 0.408g, CoCl 2 ·6H 2 O 0.06g, ZnSO 4 ·7H 2 O 0.028g, CuSO 4 ·5H 2 O 0.168g, wheat bran 6g, Tween-80 1ml, vitamin B 1 10 μg, vitamin B 2 5 μg, vitamin B 6 5 μg, add water to 1L, and sterilize at 8 pounds for 30 minutes.

[0026]Pycnoporus sanguineus (L: Fr) Murr. (MK528)) was inoculated onto solid seed medium. Culture at 30°C for 6-8 days. Get the bacterium block (diameter 1cm, 10 blocks) on the edge of the colony, inoculate into a Erlenmeyer flask filled with 100ml of high-pressure moi...

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Abstract

The invention disclosing a laccase and its production process and the special production strain, relates to the microbe-fermenting industrial sphere. The provided strain in the invention is Pycnoporus sanguineus mk528 CGMCC No. 1124. The provided strain in the invention is Pycnoporus sanguineus mk528 CGMCC No. 1124. The method for production of laccase contains fermenting and culturing Pycnoporus sanguineus mk528 CGMCC No. 1124 and extracting laccase from fermentation and culture medium. The enzyme activity of fermentation liquor laccase can reach as high as 65 U / ml by fermenting and culturing the provided Pycnoporus sanguineus mk528 CGMCC No. 1124; neither lignin peroxidase nor manganese peroxidase produced in the culture process of strain, which is convenient to extract laccase, and the extraction-acquired laccase can be largely used in paper mill wastewater disposal and substitute chemicals such as formaldehyde in wood processing industry.

Description

technical field [0001] The invention relates to the field of microbial fermentation industry, in particular to a method for producing laccase and a special bacterial strain thereof. Background technique [0002] Laccase (p-diphenol oxygen oxidoreductase, EC 1.10.3.2.) is a blue phenol oxidase containing multiple copper ions, which can catalyze the redox reaction of phenolic substances. Its reaction substrates include: phenolic and Its derivatives, aromatic amines and their derivatives, aromatic carboxylic acids and their derivatives, etc. At present, through continuous research, the range of non-phenolic substrates that can be oxidized by laccase is still expanding. Laccase can degrade phenolic and non-phenolic structures in lignin, which plays an important role in the biodegradation of lignin, and can also be used to degrade aromatic compounds. Laccase can also oxidatively polymerize lignin polyphenols in a free radical manner, which can replace synthetic resin adhesives ...

Claims

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Application Information

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IPC IPC(8): C12N1/00C12N9/02C12P21/02
Inventor 郭林王文惠张虎成李伟
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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