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Cellodextrin enzyme, encoding gene and application thereof

A technology encoding gene and cellodextrin, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of high cost, inability to truly realize industrialization, and high price, and achieve the effect of high cellodextrinase activity

Inactive Publication Date: 2010-01-20
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the wide use of cellulase and the need to use cellulase with different properties for different purposes, and because of the low efficiency and high price of cellulase, the cost of producing fuel alcohol from cellulose is too high to be truly To achieve industrialization, therefore, new cellulase enzymes are required

Method used

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  • Cellodextrin enzyme, encoding gene and application thereof
  • Cellodextrin enzyme, encoding gene and application thereof
  • Cellodextrin enzyme, encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1. Obtaining cellodextrinase Umcel5C and its coding gene

[0040] 1. Construction of a metagenomic library of uncultured microorganisms in the rumen of buffalo

[0041] 1. Extraction of metagenomics of uncultured microorganisms in the rumen of buffalo

[0042] Take 50g of buffalo rumen content (the source of the sample is the rumen of the buffalo just slaughtered in Nanning Meat Union Factory, and the collected sample is immediately stored in liquid nitrogen) and suspended in 200ml of 0.18M potassium phosphate buffer (pH6.5), Gently shake and place for 10 minutes to allow the cellulose particles in the rumen contents to automatically precipitate, discard the supernatant, and then add 200ml of 0.18M potassium phosphate buffer (pH6.5) to wash the pellet twice, the last time The precipitation block is directly extracted to extract the metagenomic DNA of the microorganisms adsorbed on the cellulose particles. Add 100ml extraction buffer (100mM sodium phosphate pH8.0; 100...

Embodiment 2

[0061] Example 2. Expression of Umcel5C in E. coli

[0062] 1. Construction of a recombinant vector (pET-umcel5C) capable of expressing umcel5C

[0063] After the gene with the coding signal peptide is expressed in vivo, the expressed product is likely to be secreted outside the cell. In order to avoid the secretion of the expression product out of the cell, it is only necessary to artificially synthesize the base from the 306th to the 1226th position of the 5′ end of the sequence 1 (that is, the amino acid in the umcel5C gene except the first 25 amino acids and the stop codon of the amino terminal of the coding sequence 2 DNA sequence, mainly to prevent the first 25 amino acids from forming a signal peptide), 921bp, the molecular weight of the encoded protein is expected to be 35.39691KDa, and the isoelectric point pI is 4.65.

[0064] Synthesize all the sequences of umcel5C gene except the first 25 amino acids and the stop codon of the amino terminal of coding sequence 2 (that is,...

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Abstract

The invention discloses a fiber dextrinase and the coding gene and the application thereof. The fiber dextrinase which is provided by the invention is a protein which has one of the following amino acid residue sequences: 1) SEQ ID No.:2 in a sequence table; 2) the protein which substitutes and / or deletes and / or adds one or more amino acid residues of the amino acid residue sequence of the SEQ ID No.:2 in the sequence table and has the fiber dextrinase activity. The experiments prove that the fiber dextrinase of the invention has very high fiber dextrinase activity (up to 42019U / g), and the appropriate pH value scope and the temperature scope of the enzyme are very broad. The fiber dextrinase and the coding gene which are provided by the invention can be widely applied in the degradation of the cellulose.

Description

Technical field [0001] The invention relates to a cellodextrinase and its coding gene and application. Background technique [0002] Cellulose is mainly the most abundant renewable biomass resource on the earth that plants use carbon dioxide and water to synthesize through photosynthesis under the action of solar energy. According to reports, the world’s annual cellulose production through photosynthesis is as high as 1.55×10 9 Tons, 89% of which have not been utilized by humans (Dunlap C, Chiang GC. Utilization and recycle of agriculture wastes and residues. Shuler M L. Boca Raton, Florida. USA: CRC Press Inc. 1980.19). Cellulose is a polymer composed of multiple glucose residues connected by β-1,4-glycosidic bonds, and its basic repeating unit is cellobiose. The basic structure of natural cellulose is made up of microfibril bundles composed of fibrils. Fibrils are composed of 15-40 long chains of cellulose molecules composed of crystalline and amorphous regions. The crystall...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12N15/56C12N15/63C12N5/10C12P19/14
Inventor 段承杰冯家勋杨伟松唐纪良
Owner GUANGXI UNIV
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