Preparation of recombinant xylanase and method for applying recombinant xylanase to cassava residue degradation

A technology of xylanase and cassava residue, which is applied in the field of bioengineering, can solve the problems of low utilization rate of lignocellulose resources, and achieve the effects of short enzyme production cycle, reduced cellulose and hemicellulose content, and easy cultivation

Active Publication Date: 2015-09-30
JIANGNAN UNIV
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  • Claims
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Problems solved by technology

[0007] Another object of the present invention is to apply the above-mentioned recombinant xylanase to treat cassava residues, improve the degradat

Method used

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Experimental program
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Effect test

Embodiment 1

[0030] The preparation of recombinant xylanase is as follows:

[0031] 1) Obtaining the target xylanase gene: using a bacterial genomic DNA extraction kit to extract the genome containing Clostridium clariflavum DSM19732 to obtain a genomic DNA solution;

[0032] 2) Construction of recombinant expression vector: according to the Clostridium clariflavum DSM19732 xylanase gene sequence reported in the database, primers with restriction endonucleases BamH I and Not I were designed, and the genomic DNA solution obtained in step 1) was used as a template for PCR Amplify and purify the amplified product, then cut the purified PCR product and plasmid pET-28a with restriction endonucleases BamH Ⅰ and Not Ⅰ, and connect with T4 ligase overnight at 16°C to obtain a recombinant expression vector ;

[0033] 3) Obtaining of recombinant xylanase: Transform the recombinant expression vector obtained in step 2) into Escherichia coli BL21 competent cells for cultivation, when OD 600 When the...

Embodiment 2

[0035] Utilize the recombinant xylanase that the present invention obtains to process cassava residue, specifically as follows:

[0036] Add 20mmol / L, pH 7.0 Tris-HCl buffer solution with a mass volume ratio of 1% to 20 mesh cassava residues, add 270U of recombinant xylanase solution and 4000U of cellulase, and keep the temperature at 55°C for 96h. Compared with the effect of cellulase treatment, the weight loss rate of cassava residue increased by 9%, the cellulose content decreased by 2.5%, and the hemicellulose content decreased by 4%.

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Abstract

The invention belongs to the technical field of bioengineering and relates to preparation of recombinant xylanase and a method for applying the recombinant xylanase to cassava residue degradation. According to the invention, the xylanase gene of Clostridium clariflavum DSM19732 is connected with plasmid pET-28a, so that a recombinant expression vector is obtained, the recombinant expression vector is transformed to escherichia coli BL21, induction expression is carried out, and cell disruption is carried out, so that the recombinant xylanase is obtained; after the recombinant xylanase is used for treating cassava residues, the contents of cellulose and hemicelluloses in the cassava residues are reduced. The recombinant xylanase prepared through the method has a good enzymatic property, and the degradation efficiency of cellulose in the cassava residues can be improved after the cassava residues are treated through the recombinant xylanase.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to the preparation of a recombinant xylanase and a method for degrading cassava residue by using the recombinant xylanase. Background technique [0002] Xylanase refers to the general term for a class of enzymes that can specifically degrade xylan into xylooligosaccharides and xylose. Since xylan is the main component of plant hemifibers, it exists widely in nature, whether as an industrial The effective degradation of xylan in raw materials or processing by-products is of great significance. Therefore, since the 1960s, people have paid attention to the research of xylanase, involving fields including food, feed, papermaking, energy industry and so on. [0003] At present, xylanase-producing bacteria include fungi, bacteria and genetically engineered bacteria. The enzymatic properties and substrate specificity of xylanase from different sources are different. The...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12N15/56C12N15/70C12P19/14C12R1/145
CPCC12N9/248C12N9/2482C12P19/14C12Y302/01008
Inventor 唐蕾胡方觊王荣余嘉慧张建华张宏建毛忠贵
Owner JIANGNAN UNIV
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