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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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The laccase activity of the known domesti

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

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[0019] Example 1. Isolation of Pycnoporus sanguineus mk528

[0020] Pycnoporus sanguineus mk528 is isolated from Pycnoporus sanguineus G5 duocytic hyphae, and Pycnoporus sanguineus G5 is a binucleated hyphae obtained from Pycnoporus sanguineus fruiting bodies (Tang Shun, Lomascolo A, Guo Lin et al. Isolation of white-rot fungi from China for screening laccase-hyperproducing strains. Bacteria system 20:520-525).

[0021] Pycnoporus sanguineus G5 contains agar 20g / l -1 And malt extract 20g / l -1 Incubate at 30°C in a petri dish. After culturing for 7 days, turn the petri dish upside down and place it at room temperature for 3-4 weeks. Then use high-temperature sterilized distilled water to collect the basidiospores from the lid of the petri dish, and then dilute these basidiospore suspensions by an appropriate multiple, and apply to agar containing 20g / l -1 And malt extract 20g / l -1 Incubate at room temperature in a petri dish. On the second or third day, separate the visible basidio...

Example Embodiment

[0022] Example 2: Fermentation and cultivation of Pycnoporus sanguineus mk528 to produce laccase

[0023] 1. Fermentation of Pycnoporus sanguineus mk528

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

[0025] Fermentation medium: 15g maltose, 2g ammonium tartrate, 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 10μg, vitamin B 2 5μg, vitamin B 6 5μg, add water to 1L, autoclave for 30 minutes at 8 lbs.

[0026]Pycnoporus sanguineus (L: Fr) Murr. (MK528)) was inoculated on the solid seed medium. Cultivate for 6-8 days at 30°C. Take the most edge of the colony (1cm in diameter, 10 pieces), inoculate it into a 100ml Erlenmeyer flask containing a fermentation medium sterilized by autoclaving, humid heat, and cultivate...

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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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