Leucobacterium laccase gene and its recombinant Pichia pastoris engineering strain and application

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 bright bacteria laccase gene and a recombinant Pichia pastoris engineering bacteria and applications thereof, and in particular to a fungal laccase gene derived from bright bacteria. Background technique

[0002] Over the past 30 years of reform and opening up, my country's economy and industry have achieved unprecedented development. However, it is an indisputable fact that industrial development has caused serious environmental pollution. Environmental pollution has become a bottleneck in 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 it is increasingly affecting the sustainable development of our national economy. At present, the waste water of our country's industries, especially the papermaking and textile printing an...

Examples

Embodiment Construction

[0027] In order to better understand the present invention, the following specific examples are given. Among them, if there is no special instructions, the methods used are all conventional methods; the materials used are all purchased from a conventional biochemical reagent store; the% in the examples, unless otherwise specified, are the mass percentages, and the quantitative test is set to three repeated experiments , The results are averaged.

[0028] 1. Material preparation

[0029] 1. The variety of bright bacteria used

[0030] The A. tabescens strain is preserved by the Institute of Biological Medicine, Zhejiang Medical College. The bright bacteria in the experiment of the present invention is a gift from Professor Ling Qingzhi of Zhejiang Medical College.

Technical field

[0001] The invention belongs to the field of biotechnology and environmental biology, and in particular relates to a bright bacteria laccase gene and a recombinant Pichia pastoris engineering bacteria and applications thereof, and in particular to a fungal laccase gene derived from bright bacteria. Background technique

[0002] Over the past 30 years of reform and opening up, my country's economy and industry have achieved unprecedented development. However, it is an indisputable fact that industrial development has caused serious environmental pollution. Environmental pollution has become a bottleneck in 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 it is increasingly affecting the sustainable development of our national economy. At present, the waste water of our country's industries, especially the papermaking and textile printing an...

Examples

Embodiment Construction

[0027] In order to better understand the present invention, the following specific examples are given. Among them, if there is no special instructions, the methods used are all conventional methods; the materials used are all purchased from a conventional biochemical reagent store; the% in the examples, unless otherwise specified, are the mass percentages, and the quantitative test is set to three repeated experiments , The results are averaged.

[0028] 1. Material preparation

[0029] 1. The variety of bright bacteria used

[0030] The A. tabescens strain is preserved by the Institute of Biological Medicine, Zhejiang Medical College. The bright bacteria in the experiment of the present invention is a gift from Professor Ling Qingzhi of Zhejiang Medical College.

Images