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

Alpha--L-rhamnosidase and application thereof

A technology of rhamnosidase and icariin, which is applied in the fields of genetic engineering and enzyme engineering, and can solve problems such as ineffective degradation

Pending Publication Date: 2018-08-31
NANJING FORESTRY UNIV
View PDF5 Cites 13 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Technical problem to be solved: Aiming at the fact that the α-L-rhamnosidase reported so far cannot effectively degrade the α-1,2 glycosidic bond linked by the natural product rhamnose, the present invention provides a kind of enzyme that can specifically degrade rhamnose α-L-rhamnosidase BtRha with Inter-α-1,2 Glycosidic Bond and Its Expression Gene and Application

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
  • Alpha--L-rhamnosidase and application thereof
  • Alpha--L-rhamnosidase and application thereof
  • Alpha--L-rhamnosidase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Cloning of α-L-rhamnosidase BtRha gene and construction of expression vector

[0030] Using the Bacteroidesthetaiotaomicron VPI-5482 genome purchased from DSMZ (https: / / www.dsmz.de / ) as a template, degenerate primers P1 and P2 were designed according to the amino acid sequence of bacterial rhamnosidase on NCBI , for the upstream and downstream primers to amplify the BtRha gene fragment, use Ex Taq polymerase (purchased from Takara Company) to prepare 50 μL reaction solution at the recommended ratio for fragment amplification, the PCR reaction conditions are 95 ° C, 5 min; 35 cycles (95 ° C, 30s; 52°C, 30s; 72°C, 2min 20s); 72°C, 10min; the reaction was stopped and kept at 4°C. PCR amplification products were purified by gel recovery kit. To obtain the Bacteroides thetaiotaomicron VPI-5482 α-L-rhamnosidase BtRha gene, the resulting PCR amplified product and pET28a were digested with Nco I and Xhol I respectively, and the fragments recovered were combined with pET-28a...

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 alpha-L-rhamnosidase and application thereof. The amino acid sequence is shown as SEQ ID NO.2. The alpha-L-rhamnosidase BtRha capable of specifically degrading alpha-1,2 glycosidic bonds is cloned and recombinantly expressed for the first time, and a blank of the alpha-L-rhamnosidase for hydrolyzing the alpha-1,2 glycosidic bonds between rhamnose is filled up. The alpha-L-rhamnosidase can efficiently and specifically degrade epmedin C and convert the epmedin C into icariin, and the molar conversion rate of 1 g / L of the epmedin C hydrolyzed by 1.5 U / mL of the alpha-L-rhamnosidase is as high as 90.5% or above under the conditions that the pH is 5.5 and the temperature is 55 DEG C.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering and enzyme engineering, and specifically relates to the expression, purification, qualitative and enzymatic preparation of icariin from α-L-rhamnosidase BtRha gene derived from Bacteroidesthetaiotaomicron VPI-5482 . Background technique [0002] α-L-rhamnosidase (α-L-rhamnosidase, EC 3.2.1.40) widely exists in animals, plants and microorganisms, and can specifically hydrolyze various glycosidic bonds, such as α-1, α-1,2 , α-1,3, α-1,4, α-1,6 glycosidic bonds, these glycosidic bonds are contained in many natural plant extract components, such as rutin, hesperidin, naringin, chaohu Glycosides such as Ding C and icariin have α-L-rhamnose groups at the ends, and the rhamnosyl groups at these ends can be hydrolyzed to improve their bioavailability, which can be used in food, medicine and other fields. The main functions and functions include: [0003] (1) Debittering effect: flavanone gl...

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): C12N9/24C12N15/56C12N15/70C12N1/21C12P19/60C12R1/19
Inventor 赵林果吴涛裴建军张珊珊段绪果
Owner NANJING FORESTRY UNIV
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