Escherichia coli engineering bacteria expressing lignin peroxidase, preparation method and application thereof

A peroxidase and Escherichia coli technology, which is applied in the field of Escherichia coli engineering bacteria for preparing and expressing lignin peroxidase, can solve the problems of difficult lignin waste biodegradable pulp bleaching, promotion and use restrictions, and unrealistic degradation. , to achieve high protein yield, facilitate gene cloning, save time and cost
CN103540606BInactive Publication Date: 2015-09-23HUNAN UNIV
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Patent Information

Authority / Receiving Office
CN Ā· China
Patent Type
Patents(China)
Current Assignee / Owner
HUNAN UNIV
Publication Date
2015-09-23
Estimated Expiration
Not applicable Ā· inactive patent

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Abstract

The invention relates to engineered escherichia coli, and also provides a preparation method of engineered escherichia coli. The preparation method comprises the following steps: cloning cDNA (complementary Deoxyribose Nucleic Acid) of a gene of phanerochaete chrysosporium lignin peroxidase; performing double digestion on the cDNA; connecting the digestion fragments with a pCold-TF vector; authenticating whether the obtained plasmid pCold-TF-LiP is subjected to positive expression; and introducing the positive plasmid pCold-TF-LiP into escherichia coli BL21 competent cells to obtain the engineered escherichia coli for expressing lignin peroxidase. The invention also provides an application of the engineered escherichia coli, namely, inoculating the engineered escherichia coli into a liquid culture medium, adding ampicillin to the liquid culture medium, culturing until OD600 (Optical Density) reaches 0.58 to 0.62, and adding a protein inductive agent IPTG (Isopropyl Thiogalactoside) to induce the expression of recombinant protein. The engineered escherichia coli is high in expression quantity and extremely high in activity, and the enzyme activity reaches 3,528U / L.
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Description

technical field

[0001] The invention relates to an Escherichia coli engineering bacterium, in particular to an Escherichia coli engineering bacterium expressing lignin peroxidase, a preparation method and application thereof. Background technique

[0002] Lignin is the second most abundant biopolymer known on the earth. It has a very complex structure and is not easy to degrade (Martinez, A. T., et al. 2005."Biodegradation of lignocellulars:microbial,chemical,and enzymatic aspects of the fungal attack of lignin." Int Microbiol 8:195-204). Due to lack of proper disposal, much lignin becomes waste, causing various environmental problems. With the increasing shortage of energy sources, researchers at home and abroad have tried to produce ethanol by converting lignin as an alternative energy source and have made some progress. Lignocellulose is also used as raw material for making paper, cultivating edible fungi and animal feed (Sanchez, C. 2009."Lignocellular residues: biodeg...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
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