Preparation method of inulin source recombinant allophycocyanin

A technology of allophycocyanin and inulin, which is applied in the field of microbial preparation of recombinant allophycocyanin from inulin sources, can solve the problems of difficulty in establishing standardized and unified quality standards, high preparation costs, and unstable effects, and achieve low cost, Energy-saving, easy-to-extract effects

Active Publication Date: 2014-03-26
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Natural phycobiliproteins not only have high preparation costs and complex processes, but also obtain pigment-protein complexes with unstable effects and it is difficult to establish a standardized and unified quality standard. In order to further study its biological activity and develop phycobiliproteins into monomer drugs Need to solve a series of costly and long-term processes such as microalgae cultivation, harvesting, phycobiliprotein extraction and purification

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Construction of strains

[0024] Escherichia coli P2 is derived from Escherichia coli strain P1 capable of producing active recombinant allophycocyanin, and constructed according to the prior art records to obtain Escherichia coli P2.

[0025] Among them, the construction method of Escherichia coli strain P1 capable of producing active recombinant phycobiliprotein is described in the literature of the application number 9612023.x, the name of the invention, the microorganism capable of producing active recombinant phycobiliprotein and its preparation method. P1 is now deposited at the General Microbiology Center (CGMCC) of the China Microorganism Collection Management Committee, with the deposit number CGMCC No. 0261, and the deposit date is April 17, 1996.

[0026] Take E. coli strain P1 as the starting strain, knock out the ampicillin resistance gene in it by molecular biological means, insert the kanamycin resistance gene in the original position, and get the kan...

Embodiment 2

[0027] Example 2 Preparation of shake flask of allophycocyanin from inulin

[0028] 1) Preparation of seed liquid: Fill the seed liquid culture medium into the Erlenmeyer flask according to the proportion of 20% (v / v) of the liquid volume, pick an appropriate amount of the above-mentioned Escherichia coli P1 slant bacteria in the inoculation loop, and then inoculate it at 37°C, Incubate for 8 hours under the condition of a shaker rotating speed of 200 rpm.

[0029] Among them, the seed medium (take the preparation of 1L medium as an example): 10g peptone, 5g yeast powder, 10g NaCl, 1L tap water, adjust the pH to 7.0 with 5mol / L NaOH solution, 115°C, sterilize for 20min, then add 1ml of 60mg / ml kanamycin solution.

[0030] 2) Preparation of allophycocyanin shake flask: Fill the fermentation medium into the Erlenmeyer flask according to the proportion of 20% of the filling volume, insert the seed solution, control the inoculum volume at 5% (v / v), and the fermentation temperature at 37...

Embodiment 3

[0035] Example 3 Fermentation tank preparation of allophycocyanin derived from inulin

[0036] 1) Preparation of the first-level seed solution: fill the seed solution medium into the Erlenmeyer flask according to the proportion of 20% (v / v) of the filling amount, pick an appropriate amount of the above-mentioned Escherichia coli P1 slant bacteria in the 37 Cultivate for 8 hours at a temperature of 200 rpm on a shaker. To

[0037] Said, the first-level seed medium (take the preparation of 1L medium as an example): 10g peptone, 5g yeast powder, 10g NaCl, 1L tap water, adjust the pH to 7.0 with 5mol / L NaOH solution, 115°C, sterilize for 20min , And then add 1ml of 60mg / ml kanamycin solution.

[0038] 2) Preparation of secondary seed liquid (take the preparation of 1L medium as an example): Fill the secondary seed liquid medium into the Erlenmeyer flask according to the proportion of 20% of the filling volume, connect the primary seed liquid, and control the inoculum 5% (v / v), and the...

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

PropertyMeasurementUnit
weightaaaaaaaaaa
Login to view more

Abstract

The invention belongs to the field of producing protein through microbiological fermentation, and in particular relates to a microbiological preparation method of inulin source recombinant allophycocyanin. The method comprises the following steps: with inulin as raw material, degrading the raw material as a seed solution, culturing Escherichia coli P2 through a shaking table or a fermentation tank under the effect of a fermentation medium, and then inductively expressing through isopropyl-beta-D-thiopyran galactoside (IPTG) to obtain the inulin source recombinant allophycocyanin. The recombinant allophycocyanin can be applied to the fields of food, healthcare and medical and functional materials. The preparation method of the inulin source recombinant allophycocyanin is simple, and an application path with low cost and high yield is found for the bio-utilization of the inulin; compared with a method of using glucose as a carbon source, the microbiological preparation method disclosed by the invention has the advantage that the raw material cost of producing the allophycocyanin in fermentation production is greatly reduced.

Description

Technical field [0001] The invention belongs to the field of protein production by a microorganism fermentation method, and specifically relates to a microorganism preparation method of inulin-derived recombinant allophycocyanin. technical background [0002] Phycobiliprotein is originally produced in the ocean and is mainly extracted from cyanobacteria and red algae. Phycobiliprotein is a functional component of the photosynthetic light-harvesting complex of cyanobacteria and red algae. According to its absorption spectrum and fluorescence spectrum characteristics, phycobiliproteins can be divided into phycoerythrin, phycoerythrin, phycocyanin and allophycocyanin protein. Phycobiliprotein is a protein with special biological properties. Phycobiliprotein can promote the regeneration of biological blood cells, comprehensively enhance the body's disease prevention and disease resistance, and can inhibit the growth of cancer cells. It is an ideal photosensitizer with no toxic side...

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): C12P21/02C12R1/19
Inventor 李莉莉秦松席海瑞张锐宋璐非王绍宇刘冰
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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