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Cellodextrin enzyme, encoding gene and application thereof

A technology encoding gene and dextrinase, applied in application, genetic engineering, plant gene improvement and other directions, can solve the problems of high cost, low efficiency and high price, and achieve the effect of high cellodextrinase activity

Inactive Publication Date: 2008-07-16
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] Embodiment 1, the acquisition of cellodextrinase Umcel5C and its coding gene

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

[0041] 1. Extraction of uncultured microbial metagenomics from buffalo rumen

[0042] Get 50g buffalo rumen contents (the source of the sample is the rumen of the buffalo just slaughtered by Nanning Meat Co., Ltd., and the collected sample is immediately preserved with liquid nitrogen), suspended in 200ml of 0.18M potassium phosphate buffer (pH6.5), Gently oscillate evenly, let stand for 10 minutes, let the cellulose particles in the rumen content automatically precipitate, discard the supernatant, and then add 200ml of 0.18M potassium phosphate buffer (pH6.5) to wash the sediment twice, and the last time Metagenomic DNA of microorganisms adsorbed to cellulose pellets was extracted by direct extraction from pellets. Add 100ml of extraction buffer (100mM sodium phosphate pH8.0; 100mM Tirs-HCl pH8.0; 10...

Embodiment 2

[0061] Embodiment 2, the expression of Umcel5C in escherichia coli

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

[0063] After the gene with coding signal peptide is expressed in vivo, the expression product is likely to be secreted outside the cell. In order to prevent the expression product from being secreted outside the cell, only the 306th to 1226th base of the 5' end of the artificially synthesized sequence 1 (that is, the first 25 amino acids of the amino terminal of the coding sequence 2 and the stop codon in the umcel5C gene are excluded. The DNA sequence is mainly to prevent the first 25 amino acids from forming a signal peptide), 921bp, the molecular weight of the encoded protein is estimated to be 35.39691KDa, and the isoelectric point pI is 4.65.

[0064] Artificially synthesized the entire sequence of the umcel5C gene except the first 25 amino acids at the amino terminal of the coding sequence 2 and the stop codon ...

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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, its encoding gene and application. Background technique [0002] Cellulose is mainly the most abundant renewable biomass (biomass) resource on earth synthesized by plants using carbon dioxide and water through photosynthesis under the action of solar energy. According to reports, the global annual production of cellulose 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-glucosidic bonds, and its basic repeating unit is cellobiose. The basic structure of natural cellulose is composed of microfibril bundles composed of fibrils. Fibrils are composed of 15-40 long chains of cellulose molecules with crystalline and non-crystalline regions. T...

Claims

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Application Information

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Patent Type & Authority Applications(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