Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Acidophil Beta-glucanase GLU7A and gene and application thereof

A glucanase and gene technology, applied in the field of genetic engineering, can solve problems such as differences in glucanase activity, achieve high enzyme activity stability, reduce environmental pollution, and increase productivity.

Active Publication Date: 2010-06-23
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
View PDF0 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Genes from different sources can be expressed in different vectors and host systems, including Escherichia coli, Bacillus, Saccharomyces cerevisiae, and transgenic barley and tobacco, but the glucanase activity of different expression systems There is a big difference

Method used

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
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Acidophil Beta-glucanase GLU7A and gene and application thereof
  • Acidophil Beta-glucanase GLU7A and gene and application thereof
  • Acidophil Beta-glucanase GLU7A and gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Acidophilic fungus Bispora sp. MEY-1 and its enzyme production characteristics

[0039] A sample of uranium ore wastewater from a mine in Jiangxi was cultured in potato juice medium, and then coated on an enzyme-producing medium ((NH 4 ) 2 SO 4 5g / L, KH 2 PO 4 1g / L, MgSO 4 ·7H 2 O 0.5g / L, FeSO 4 ·7H 2 O 0.01g / L, CaCl 2 0.2g / L, dextran 1%, 1.5% agarose, pH 2.5) plate, cultured at 30°C for 5-6 days, and transparent circles can be seen on the enzyme production medium plate. Prove that it has glucanase activity.

[0040] The acidophilic fungus Bispora sp.MEY-1 was deposited in the General Microbiology Center of the China Microbial Culture Collection Management Committee (Datun Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, 100101) on May 19, 2008, and its collection number It is: CGMCC No.2500.

Embodiment 2

[0041] Example 2 Cloning of glu7A encoding gene glu7A from the acidophilic fungus Bispora sp.MEY-1

[0042] Extract genomic DNA from the acidophilic fungus Bispora sp.MEY-1:

[0043] Filter the mycelium cultured for 3 days with sterile filter paper and put it in a mortar, add 2 mL of extract, grind for 5 min, then place the grind in a 50 mL centrifuge tube, lyse in a water bath at 65°C for 20 min, and mix every 10 min. Homogenize once and centrifuge at 10,000 rpm at 4°C for 5 min. The supernatant was extracted in phenol / chloroform to remove impurities, and then the supernatant was added to an equal volume of isopropanol. After standing at room temperature for 5 min, centrifuged at 10,000 rpm at 4°C for 10 min. The supernatant was discarded, the precipitate was washed twice with 70% ethanol, dried in vacuum, dissolved in an appropriate amount of TE, and placed at -20°C for use.

[0044] Degenerate primers P1: 5'-ggYTACTgYgAYgCNCARTg-3'; and P2: 5'-TAgggRTTgAANCCRCANCC-3' were design...

Embodiment 3

[0049] Example 3 RT-PCR analysis of glucanase gene

[0050] Extract the total RNA of Bispora sp.MEY-1, use reverse transcriptase to obtain a strand of cDNA, and then design appropriate primers (glu7AF: 5′-ATGGCAGTGTTCACTCTGACCGTGG-3′, glu7AR: 5′-CTAATTCCAACCGCCAGAGCC-3′) to amplify From the single-stranded cDNA, a cDNA sequence encoding a glucanase is obtained, and the amplified product is recovered and sequenced.

[0051] By comparing the genomic sequence and cDNA sequence of glucanase glu7A, it is found that the gene has a 60bp intron. The full length of the cDNA is 1281bp, which encodes 427 amino acids. The first 19 amino acids at the N-terminal are possible signal peptide sequences. The BLAST alignment results showed that it had the highest sequence similarity with endo-1,4-beta-glucanase derived from Aspergillus oryzae. The highest similarity of amino acids is 58%; the highest similarity of nucleotides is 60.3%. It proved that the gene encoding glucanase isolated and cloned ...

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
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of genetic engineering, in particular to an acidophil Beta-glucanase GLU7A and gene and application thereof. The invention provides a glucanase GLU7A from acidophil bispora Bisporasp, and the amino acid sequence thereof is shown as SEQIDNO.1, and the invention provides a genome of coding the glucanase and cDNA coding gene glu7A. The invention obtains an acidophil glucanase which has the optimum pH value of 1.5-5.0, simultaneously remains high enzymatic activity in acid environment, and resists protease hydrolysis. In addition, the glucanase not only hydrolyzes Beta-1, 3-1, 4-glucanase, but also can hydrolyze cellulose. Due to the properties, the acidophil Beta-glucanase GLU7A can be applied in the industries of food, feedstuff and beer.

Description

Technical field [0001] The present invention relates to the field of genetic engineering. Specifically, the present invention relates to an acidophilic β-glucanase GLU7A and its gene, a recombinant vector containing the gene, and applications. Background technique [0002] β-glucan is a structural non-starch polysaccharide in the cell walls of higher plants in the Gramineae family, and is particularly abundant in the endosperm cell walls of cereals (barley, oats, sorghum, rice and wheat). [0003] β-glucan is a polysaccharide polymer linked by D-type β-conformation glucose and has a linear spatial structure. According to the different glycosidic bond connection, β-glucan can be divided into 1,4-β-glucan, 1,3-β-glucan, 1,6-β-glucan, 1,3-1 , 6-β-glucan, 1,3-1,4-β-glucan, etc. Among them, 1,3-1,4-β-glucan is connected by β-1,3 and β-1,4 mixed glycosidic bond, usually by β-1,4 glycosidic bond to connect glucose to form cellotriose or Cellotetraose is connected to each other by β-1,3...

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
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12N15/56C12N15/63C12N1/21C12N1/19C12P19/14C12R1/225C12R1/07C12R1/645C12R1/19
Inventor 姚斌罗会颖王亚茹杨培龙孟昆史秀云柏映国袁铁铮石鹏君杨君
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com