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

Application of protein CGTase as cyclodextrin glycosyltransferase

A glycosyltransferase, protein technology, applied in the direction of glycosyltransferase, transferase, application, etc., can solve the problems of small production scale, limited production and application of γ-CD, and high price

Pending Publication Date: 2021-12-17
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

γ-CD is also highly safe and can be quickly and basically completely digested by salivary amylase and pancreatic isoamylase in the human body. Therefore, γ-CD can be rapidly degraded and absorbed in the human small intestine, but α-CD and β- CDs cannot
The application prospect of γ-CD is broader than that of β-CD, but there is currently no γ-CGTase with high specificity, resulting in a small production scale and high price, which limits the production and application of γ-CD

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
  • Application of protein CGTase as cyclodextrin glycosyltransferase
  • Application of protein CGTase as cyclodextrin glycosyltransferase
  • Application of protein CGTase as cyclodextrin glycosyltransferase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Embodiment 1, the discovery of protein CGTase1 and its 15 mutant proteins

[0097] 1. Discovery of the CGTase1 gene

[0098] The CGTase1 gene shown in SEQ ID NO: 2 was artificially synthesized.

[0099] The CGTase1 gene encodes the protein CGTase1 shown in SEQ ID NO:1.

[0100] 2. Obtaining Mutant Proteins

[0101] The inventors of the present invention carried out saturation mutation at position 248 of protein CGTase1, and obtained 15 mutant proteins including deletion mutations. details as follows:

[0102] (1) Deleting the 742-744 nucleotides (GCA) from the 5' end of SEQ ID NO: 2 to obtain double-stranded DNA molecule 1. The protein encoded by double-stranded DNA molecule 1 is named protein 1.

[0103] That is, protein 1 is obtained after the alanine at position 248 of protein CGTase1 is deleted.

[0104] (2) Replace the 742nd-744th nucleotides (GCA) of SEQ ID NO: 2 from the 5' end with CGA to obtain double-stranded DNA molecule 2. The protein encoded by the d...

Embodiment 2

[0132] Example 2. Application of the protein CGTase1 and its muteins as cyclodextrin glycosyltransferases in the production of α-CD, β-CD and γ-CD

[0133] 1. Construction of recombinant plasmid 1-recombinant plasmid 16

[0134] 1. Artificially synthesize the double-stranded DNA molecule 1-double-stranded DNA molecule 15 in Step 2 of Example 1 and the double-stranded DNA molecule shown in SEQ ID NO: 2 (named double-stranded DNA molecule 16).

[0135] 2. Using double-stranded DNA molecule 1 as a template, use primer S2: 5'-CGC GGATCC ATGATTCGAAGGCTTTC-3' (the underline is the recognition sequence of restriction endonuclease BamHI) and primer A3: 5'-CGG CTCGAG A primer pair consisting of TTGATTGTAATTCACTTC-3' (the underline is the recognition sequence of the restriction endonuclease XhoI) was used for PCR amplification to obtain a PCR amplification product of about 2000 bp.

[0136] 3. Digest the PCR amplified product obtained in step 2 with restriction endonucleases BamHI a...

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 discloses an application of a protein CGTase as a cyclodextrin glycosyl transferase. The protein CGTase sequentially comprises a segment 1 to a segment 3 from an N terminal to a C terminal, the amino acid sequence of the segment 1 is as shown in SEQ ID NO: 1 from the 1st site to the 247th site from the N terminal, the segment 2 is an amino acid residue, and the amino acid sequence of the segment 3 is as shown in SEQ ID NO: 1 from the 249th site to the 699th site from the N terminal. Experiments prove that the protein CGTase has cyclodextrin glycosyl transferase activity and can degrade starch. The method has an important application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of protein CGTase as cyclodextrin glycosyltransferase. Background technique [0002] Cyclodextrin (CD) is produced by cyclodextrin glycosyltransferase (cyclodextrin glucano-transferase, referred to as CGTases) and starch cyclization, which is composed of six or more glucose units through α-(1,4) A class of cyclic oligomeric compounds linked by glycosidic bonds. The more common ones are α-cyclodextrin (α-cyclodextrin, α-CD), β-cyclodextrin (β-cyclodextrin, β-CD) and γ-cyclodextrin (γ-cyclodextrin, γ-CD), respectively Consisting of 6, 7 and 8 glucose unit molecules, these three cyclodextrins are the most widely used three cyclodextrins in industry. [0003] Cyclodextrins catalyze starch production mainly by cyclodextrin glycosyltransferases. The generated products are mainly three mixtures of α-CD, β-CD and γ-CD, and the product specificity is poor. The ...

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
IPC IPC(8): C12N9/10C12N15/54C12P19/18C12P19/04
CPCC12N9/1074C12P19/18C12P19/04C12Y204/01019
Inventor 钞亚鹏王国林范婷文
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF 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