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

Method for improving enzyme activity of trehalose synthase by C-terminal fragments of thermophilic bacteria trehalose synthase

A technology of trehalose synthase and thermophilic bacteria, applied in the field of enzyme genes, can solve problems such as improving trehalose production

Active Publication Date: 2015-05-27
BEIJING UNIV OF CHEM TECH
View PDF1 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In summary, the expression of trehalose synthase genes from four sources, Pseudomonas putida, Corynebacterium glutamicum, Streptomyces coelicolor and Thermotoga maritima, in Escherichia coli has not yet been combined with Fusion expression method of the C-terminal fragment of trehalose synthase of thermobacteria to improve its trehalose production

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
  • Method for improving enzyme activity of trehalose synthase by C-terminal fragments of thermophilic bacteria trehalose synthase
  • Method for improving enzyme activity of trehalose synthase by C-terminal fragments of thermophilic bacteria trehalose synthase
  • Method for improving enzyme activity of trehalose synthase by C-terminal fragments of thermophilic bacteria trehalose synthase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Example 1 Cloning of the coding region of the trehalose synthase gene derived from Pseudomonas putida, Corynebacterium glutamicum, Streptomyces coelicolor and Thermotoga maritima and its fusion expression in Escherichia coli

[0089] 1. Primer design and PCR reaction

[0090] Pseudomonas putida NBRC14164, Corynebacterium glutamicum ATCC 13032, Streptomyces coelicolor ATCC 23899, Thermotoga marine MSB8 and Thermus thermophilus) HB27 genome was provided by Yan Yajun, a professor at the University of Georgia in the United States, and these strains can be purchased from the China General Microorganism Culture Collection Center.

[0091] According to the known coding sequence of trehalose synthase gene from four sources of Pseudomonas putida, Corynebacterium glutamicum, Streptomyces coelicolor and Thermotoga maritima, a pair of upstream and downstream primers were designed respectively. for:

[0092] The upstream primer of the trehalose synthase gene fragment derived from ...

Embodiment 2

[0145] Example 2 The C-terminal thermophilic fragment gene coding region of thermophilic bacteria trehalose synthase derived from Pseudomonas putida, Corynebacterium glutamicum, Streptomyces coelicolor and Thermotoga maritima fused to it Cloning and fusion expression in Escherichia coli

[0146] 1. Primer design and PCR reaction

[0147] Pseudomonas putida NBRC14164, Corynebacterium glutamicum ATCC 13032, Streptomyces coelicolor ATCC 23899, Thermotoga marine MSB8 and Thermus thermophilus) HB27 genome was provided by Yan Yajun, a professor at the University of Georgia in the United States, and these strains can be purchased from China General Microorganism Culture Collection Center.

[0148] According to the sequence of the C-terminal thermophilic fragment coding region of trehalose synthase gene from known thermophilic bacteria and seaweed from four sources: Pseudomonas putida, Corynebacterium glutamicum, Streptomyces coelicolor and Thermotoga maritima Sugar synthase gene co...

Embodiment 3

[0224] Example 3 Effect of C-terminal fragment of trehalose synthase derived from thermophilic bacteria on enzymatic activity of trehalose synthase derived from Pseudomonas putida, Corynebacterium glutamicum, Streptomyces coelicolor and Thermotoga maritima

[0225] The enzymatic activity and activity of four trehalose synthases derived from Pseudomonas putida, Corynebacterium glutamicum, Streptomyces coelicolor and Thermotoga maritima by adding the C-terminal gene fragment of trehalose synthase derived from thermophilic bacteria Compared with the original trehalose synthase, it was increased by 2.53 times, 5.58 times, 2.74 times and 4.57 times respectively. Therefore, it is proved that the trehalose synthase derived from thermophilic bacteria does have a certain impact on the catalytic properties of the other four sources of trehalose synthase, and plays a crucial role.

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 a method for improving enzyme activity of trehalose synthase by C-terminal fragments of thermophilic bacteria trehalose synthase. Four C-terminal fragments of trehalose synthase from different sources connected with thermophilic bacteria-originated C-terminal gene fragments of trehalose synthase are subjected to fusion expression in escherichia coli while expressing trehalose synthase originated from pseudomonas putida, corynebacterium glutamicum, streptomyces coelicolor and thermotoga maritime by applying a molecular biological technique. Under the action of the thermophilic bacteria-originated C-termianl gene fragments of trehalose synthase, the enzyme catalytic activity of trehalose synthase is improved. According to the method disclosed by the invention, modified trehalose synthase is expressed in escherichia coli, and thus the enzyme activity of trehalose synthase is improved.

Description

technical field [0001] The invention belongs to the field of biochemistry, in particular to trehalose synthase gene derived from thermophilic bacteria and its C-terminal truncated fragment, to trehalose synthase gene derived from Pseudomonas putida and its truncated fragment, to glutamic acid The invention relates to trehalose synthase genes from three sources of corynebacterium, streptomyces coelicolor and thermotoga maritima, and also relates to recombinant expression vectors and applications containing these trehalose synthase genes. Background technique [0002] Trehalose is a non-reducing disaccharide formed by connecting two molecules of glucose with 1,1-glycosidic bonds. It is widely found in organisms such as bacteria, yeast, filamentous fungi, plants, insects, and invertebrates. Trehalose has very important biological significance to organisms, and is a stabilizer and protector for proteins and biofilm molecules in harsh environments such as dehydration, high temper...

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): C12N9/10
CPCC12N9/90C12Y504/99016
Inventor 袁其朋李妍冯越
Owner BEIJING UNIV OF CHEM TECH
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