Fungal laccase mutant PIE5 as well as expression strain and application thereof

A fungal laccase and mutant technology, applied in fungi, enzymes, oxidoreductases, etc., can solve the problems of inactivity, unfavorable treatment, environmental protection, and increased washing costs.

Active Publication Date: 2019-02-01
ANHUI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

The technical effect that this innovation solves by improving its performance over existing methods.

Problems solved by technology

Laccases from different genera found throughout nature produce hydroxycinnamyl alcohydroquinone compounds called lacillin A which plays an essential role for many metabolism processes including cellular signal transduction pathways. These enzymes play crucial roles during respiration by converting chemicals into energy needed for cells like mitochondria. They work with other organics containing sulfur groups attached to their side chains. However, these soluble metalloproteins cannot perform well when exposed alone due to low stability towards strong acids and bases. To address this problem, researchers developed methods to enhance lytic properties of certain types of lactas produced naturally while reducing unwanted effects on others involved in industrial production.

Method used

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  • Fungal laccase mutant PIE5 as well as expression strain and application thereof
  • Fungal laccase mutant PIE5 as well as expression strain and application thereof
  • Fungal laccase mutant PIE5 as well as expression strain and application thereof

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Embodiment Construction

[0029]The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art. In the following examples, various processes and methods not described in detail are conventional methods well known in the art.

[0030] 1. Preparation of fungal laccase PIE5

[0031] Strains: E.coli JM109, P.pastoris GS115

[0032] Vector: pPIC9K

[0033] Medium:

[0034] YPD medium (1L): 20g Peptone, 10g Yeast Extract dissolved in 900mL water, after autoclaving, add 100mL 10×D, add 1.5% agar to the solid medium.

[0035] The solvent of the seed liquid medium is PBS buffer solution with a pH value of 6.0 and 0.1M, and the solute and its concentration are as follows: YeastExtract 10g / L, Tryptone 20g / L, YNB 13.4g / L, glycerol 10g / L, biotin 4×10 -4 g / L.

[0036] The solvent of the fermentation liquid medium is PBS buffer solution with a pH value of 6.0 and 0.1M, and the solute and its concentration are as follows: YNB13.4 g / ...

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Abstract

The invention discloses a fungal laccase mutant PIE5 as well as an expression strain and application thereof, wherein the fungus laccase mutant PIE5 is a laccase mutant obtained by carrying out transformation on Coprinus cinereus laccase LCC 9 by virtue of a directed evolution means, constructing a mutation library and combining three rounds of screening, and the application capability of the laccase can be improved. The optimal catalytic pH of the fungal laccase mutant PIE5 is 8.5 (guaiacol is used as a substrate), and the fungal laccase mutant PIE5 can still play good activity when the pH value reaches 9.5, so that the fungal laccase mutant PIE5 has an application advantage in an alkaline environment, the application field of the fungal laccase is expanded, the fungal laccase mutant PIE5can be directly applied to a textile finishing process to replace stone mill water washing, the pH of wastewater does not need to be adjusted in a wastewater indigo degradation process, improvement of the textile finishing process of the fungal laccase is facilitated, the operation cost is reduced, and the ecological environment is protected.

Description

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Claims

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Application Information

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Owner ANHUI UNIVERSITY
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