Recombinant bacillus subtilis expressing C30 carotenoid

A technology of Bacillus subtilis and carotene, applied in the field of biotechnology and genetic engineering, to achieve the effect of reducing the symptoms of colitis

Active Publication Date: 2016-08-10
NANJING AGRICULTURAL UNIVERSITY
View PDF2 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But this enzyme has not been identified in bacteria

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
  • Recombinant bacillus subtilis expressing C30 carotenoid
  • Recombinant bacillus subtilis expressing C30 carotenoid
  • Recombinant bacillus subtilis expressing C30 carotenoid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0083] 1 Construction of carotenoid synthase plasmid pMK3-crtmn

[0084] The methods used in the following examples are conventional methods unless otherwise specified. The specific materials and reagents involved are as follows:

[0085] Bacillus subtilis strain WB800 was donated by Gao Xuewen, School of Plant Protection, Nanjing Agricultural University. For literature, see: Wu, H., Wang, S., Qiao, J., Liu, J., Zhan, J., Gao, X., 2009. Expression of HpaG Xooc proteinin Bacillus subtilis and its biological functions. Journal of Microbiology and Biotechnology 19, 194-203;

[0086] E.coli JM109, plasmids pMK3, pCDNA3.1 (purchased from Life Technologies);

[0087] Reagents: Q5 high-fidelity enzyme was purchased from NEB Company; pMD19-T vector, Agarose Gel DNA Purification Kit, restriction endonuclease, T4 ligase, etc. were purchased from Takara Company; kanamycin sulfate, ampicillin, bacterial genome extraction reagents The kit was purchased from OMEGA; the One Step Cloning...

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 recombinant bacillus subtilis capable of synthesizing carotenoid 4,4'-diaponeurosporene with 30 carbon atoms (C30), and belongs to the biotechnology genetic engineering field. A bacillus subtilis expression plasmid pMK3-crtmn (which is capable of expressing C30 carotenoid synthases Crtm and Crtn in bacillus subtilis) containing C30 carotenoid synthase genes crtm and crtn is successfully constructed, and is successfully electro-transformed into bacillus subtilis WB800. The WB800 strain after transformation turns from the original color of white to yellow. A pigment component extracted from the recombinant WB800 is identified as 4,4'-diaponeurosporene by high performance liquid chromatography (HPLC). 4,4'-diaponeurosporene can stimulate maturation of dendritic cells, and generated cytokines (IL-6, IL-10, IL-12p70 and TNF[alpha]) have the amount increased, and moreover and have stronger ability to stimulate proliferation of T lymphocytes. With oral administration of the bacillus subtilis capable of synthesizing 4,4'-diaponeurosporene, the dextran sulphate sodium (DSS)-induced mice colitis symptoms can be significantly reduced. The bacillus subtilis capable of producing 4,4'-diaponeurosporene is expected to be developed as probiotics for prevention of colitis.

Description

1. Technical field [0001] The present invention relates to the expression of C 30 The invention relates to carotenoid recombinant bacillus subtilis, which belongs to the field of biotechnology genetic engineering. Specifically include: C 30 Construction of carotenoid expression plasmid pMK3-crtmn; C 30 Biosynthesis and identification of carotenoids; C 30 Carotenoids can stimulate the maturation of dendritic cells, increase the amount of cytokines (IL-6, IL-10, IL-12p70 and TNFα) produced, and have a stronger ability to stimulate the proliferation of T lymphocytes; oral administration can express C 30 Bacillus subtilis carotenoids can significantly reduce dextran sodium sulfate (DSS)-induced colitis symptoms in mice. 2. Background technology [0002] 1. Research progress on carotenoid synthesis [0003] Carotenoids are a class of substances that widely exist in nature, many of which are essential nutrients for humans and animals. Carotenoids in nature come from two carb...

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): C12N15/75C12N1/21C12Q1/02A61K35/742A61P1/00C12R1/125
CPCA61K35/742C12N9/00C12N15/75C12N2800/101G01N33/5038G01N33/5047
Inventor 杨倩刘浩飞徐文雯朱立麒
Owner NANJING AGRICULTURAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap