Application of bovine rumen microorganism cellulase eg gene in lactobacillus expression

Abstract

The invention discloses an application of a bovine rumen microorganism cellulase eg gene in lactobacillus expression, and belongs to the technical field of bioengineering. The method comprises the following steps: extracting bovine rumen total DNA (Deoxyribose Nucleic Acid), carrying out PCR (Polymerase Chain Reaction) amplification on an eg fragment by taking an eg gene sequence of NCBI (National Center of Biotechnology Information) as a design primer and the bovine rumen total DNA as a template, and cloning the eg fragment into an expression vector pMG36e to obtain a secretory expression vector pMG36e::eg; and transferring the recombinant plasmid subjected to correct sequencing electrically into the L.lactis NZ9000, so that the L.lactis NZ9000 / pMG-36e::eg recombinant strain is obtained. According to the invention, a cellulase eg gene of bovine rumen is cloned, and a secretory expression plasmid pMG36e:: eg is successfully constructed by connecting typing expression plasmids. A strip with a gene size of 1500bp is cloned from a bovine rumen, the molecular weight of the enzyme is 50kD, the congo red dyeing has obvious hydrolysis rings, and after the enzyme activity is determined by a DNS method, the enzyme activity value is 1.8195 U / mL, which indicates that the enzyme has certain activity.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to an intelligent power Internet of Things monitoring platform. Background technique [0002] Cellulose is the main component of plant cell walls. Cellulose is the oldest and most abundant natural polymer on the earth. It is inexhaustible and the most precious natural renewable resource for human beings. With the development of animal husbandry in my country, straw is the material with the most cellulose content. Cellulose can usually be treated by physical and chemical methods, usually by incineration, which not only pollutes the environment, but also makes the resources inefficient due to the complicated industrial process. use. [0003] Cellulase, as an enzyme that can degrade cellulose, generally exists in plants and degrades plant cell walls. However, this enzyme has a small content in plants and can only be obtained from animals or microorganisms. It has bee...

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Problems solved by technology

With the development of animal husbandry in my country, straw is the material with the most cellulose content. Cellulose can usually be treated by physical and chemical methods, usually by incineration, which not only pollutes the environment, but also makes the resources inefficient due to the complicated industrial process. use

[0003] Cellulase, as an enzyme that can degrade cellulose, generally exists in plants and degrades plant cell walls. However, this enzyme has a small content in plants and can only be obtained from animals or microorganisms. It has been found that cellulose can be produced Enzyme microorganisms include bacteria, fungi and actinomycetes, etc. Cellulase produced by bacteria has no activity on crystalline cellulose

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