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Armillariella tabescens laccase gene, recombined pichia pastoris engineering bacteria thereof and application

A Pichia pastoris, engineering bacteria technology, applied in genetic engineering, application, plant genetic improvement and other directions, can solve the problems of high cost of lignocellulose-degrading enzymes and lignin-degrading enzymes, inability to meet industrial needs, and low yields.

Active Publication Date: 2016-05-04
GUANGXI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that lignocellulose degrading enzymes and lignin degrading enzymes are high in cost and low in yield, which cannot meet industrial needs

Method used

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  • Armillariella tabescens laccase gene, recombined pichia pastoris engineering bacteria thereof and application
  • Armillariella tabescens laccase gene, recombined pichia pastoris engineering bacteria thereof and application
  • Armillariella tabescens laccase gene, recombined pichia pastoris engineering bacteria thereof and application

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Embodiment Construction

[0027] For a better understanding of the present invention, the following are specifically described by examples. Wherein, unless otherwise specified, the methods used are conventional methods; the materials used are all purchased from conventional biochemical reagent stores; the % in the examples, unless otherwise specified, are mass percentages, and the quantitative tests are all set to repeat the experiment three times , and the results are averaged.

[0028] 1. Material preparation

[0029] 1. The species of bright bacteria used

[0030] Bright bacteria (A. tabescens) strains were preserved by the Institute of Biopharmaceuticals of Zhejiang Medical College. The bright bacteria in the experiment of the present invention was donated by Professor Ling Qingzhi of Zhejiang Medical College.

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Abstract

The invention discloses armillariella tabescens laccase and a gene thereof. The inventor constructs recombined pichia pastoris engineering bacteria according to armillariella tabescens laccase and the gene thereof. By using the engineering bacteria, efficient fermentation can be achieved for obtaining recombined armillariella tabescens laccase. On the basis of pichia pastoris heterologous expression, the inventor carries out purification and enzymology activity study on zymoprotein. The result shows that laccase has laccase activity, armillariella tabescens laccase or recombined armillariella tabescens laccase can catalyze 2,4-dichlorophenol degradation effectively, metallic copper ions have a function of promoting laccase activity, while manganese ions, zinc ions, magnesium ions, calcium ions, silver ions and SDS have a function of inhibiting activity. Stability analysis of prepared recombined laccase shows that stability of recombined laccase is good, acid environment tolerance is high, and application value is high.

Description

technical field [0001] The invention belongs to the field of biotechnology and environmental biology, and in particular relates to a laccase gene of Leucobacterium spp. and its recombinant Pichia pastoris engineering bacteria and its application, in particular to the laccase gene of fungi derived from sclerobacteria. Background technique [0002] Over the past 30 years of reform and opening up, my country's economy and industry have achieved unprecedented development, but it is an indisputable fact that industrial development has caused serious environmental pollution. Environmental pollution has become a bottleneck of economic development and a threat to human survival. It has also become an extremely important issue in current international relations and economic and trade cooperation, and has increasingly seriously affected the sustainable development of my country's national economy. At present, my country's industrial waste water, especially the papermaking and textile ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/02C12N1/19C12N15/81A62D3/02A62D101/28
CPCA62D3/02A62D2101/28C12N9/0061C12Y110/03002
Inventor 詹洁赵帅凌庆枝范业赓
Owner GUANGXI UNIV
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