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

Method for sifting and producing generation agent of dual-rhamnolipid

A double rhamnolipid and screening method technology, which is applied in the field of screening and preparation of double rhamnolipid producing bacteria, can solve the problems of poor selectivity, heavy workload, low yield of double rhamnolipid, etc., and achieve selectivity Strong, reduce workload, improve the effect of screening accuracy

Inactive Publication Date: 2008-05-07
NANJING UNIV OF SCI & TECH
View PDF0 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The above-mentioned method for preparing double rhamnolipids has the following deficiencies: (1) no matter screening bacterial strains or adopting existing bacterial strains, the output of rhamnolipids can not be guaranteed as the basis for selection, and the result show that the double rhamnolipid production is indeed very low
Therefore, this method has poor selectivity and heavy workload

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 sifting and producing generation agent of dual-rhamnolipid
  • Method for sifting and producing generation agent of dual-rhamnolipid
  • Method for sifting and producing generation agent of dual-rhamnolipid

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0028] In conjunction with Fig. 1, the production bacteria screening and preparation method of double rhamnolipids of the present invention comprises the following steps:

[0029] a. Select the soil samples collected on-site polluted by oil as the strain screening source;

[0030] b. After soaking the sample, inoculate the enrichment medium with vegetable oil as the carbon source, and culture it for 24-72 hours to obtain the enrichment medium;

[0031] c. Gradiently dilute the enriched culture solution and spread it on the hexadecyltrimethylammonium bromide plate culture medium for 24-96 hours, and pick a single colony with good growth and a large number to preserve the species on the slant;

[0032] d. Pick the bacterial strains preserved on the slant and insert them into the fermentation medium for 48-120 hours to measure the surface activity. Select the supernatant of the bacterial strains with higher activity for TLC analysis. The developing agent is chloroform: methanol: ...

Embodiment 1

[0039] Embodiment 1: in conjunction with Fig. 2, the screening method for the producing bacteria of double rhamnolipids of the present invention, promptly collect soil samples near the kitchen sewage outlet polluted by grease for a long time, weigh 10g soil samples and add to the beaker that 50mL deionized water is housed In, stir to mix. After soaking for 1 hour, take 20mL and insert it into 80mL enrichment medium, put it in a 250mL Erlenmeyer flask, and culture it on a shaker at 32°C with a rotation speed of 230r / min. After 36 hours, the enrichment solution was taken out, and serially diluted, and the dilution factor was taken as 10 4 , 10 5 , 10 6 Coat the cetyltrimethylammonium bromide agar medium with the bacterial suspension, place it in an incubator, and incubate at 30°C for 48 hours (it is also acceptable if the ambient temperature is lower than this temperature, but it is necessary to prolong the cultivation time until the colony grows to It is easy to observe the ...

Embodiment 2

[0040] Embodiment 2: The method for screening the bacteria producing double rhamnolipids of the present invention is to collect soil samples near the exhaust pipe of the range hood, weigh 10 g of the soil samples and add them to a beaker filled with 50 mL of deionized water, and stir to mix them evenly. After soaking for 1 hour, take 10mL and insert it into 40mL enrichment medium, put it in a 150mL Erlenmeyer flask, and place it in a shaker at no higher than 40°C, with a rotation speed of 230r / min. After 24-72h, the enrichment solution was taken out and serially diluted, and the dilution factor was taken as 10. 3 , 10 4 , 10 5 Coat cetyltrimethylammonium bromide agar medium with the bacterial suspension, place in an incubator, incubate at 40°C for 24-96 hours, observe the colony shape, and select the strains with the majority of colonies of the same shape, especially Colonies with the following characteristics: off-white, flat and amorphous, diffuse or slightly spread to 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
Diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a method for screening and preparing culture seed for dirhamnolipids, which comprises the following steps: choosing the soil sample polluted by grease chronically as the strain source in strain screening; adopting vegetable oil as the liquid culture medium of single carbon source, and coating with the vegetable oil on the hexadecyl trimethyl ammonium bromide agar plate with stronger selectivity after enrichment; then selecting suspect strain depending on the morphology and growth vigour of the bacterial colony; testing the surfactivity of the strain under primary screening after fermentation culture; analyzing the fermentation broth of the strain with high activity directly with TLC method; then selecting the strain with high dirhamnolipids productivity; extracting the rhamnolipids from the strain fermentation broth under screening with extraction method; obtaining purer dirhamnolipids with column chromatographic separation method. The invention overcomes the disadvantage that the strain is screened only based on the total output of rhamnolipids, and the strain with high dirhamnolipids output is obtained difficultly in the prior screening method, and has the advantages of high accuracy, high rhamnolipids output of the screened strain, more particularly higher dirhamnolipids output, and simple preparation technology.

Description

1. Technical field [0001] The invention relates to a preparation method of a biosurfactant with medicinal value, in particular to a screening and preparation method of dirhamnolipid-producing bacteria. 2. Background technology [0002] Rhamnolipids are a class of surface active substances produced by microbial metabolism. In addition to the general functions of chemically synthesized surfactants, rhamnolipid products are non-toxic, environmentally friendly, and can be 100% degraded by microorganisms, and can be widely used in many fields of industry, agriculture and daily life. Recent studies have found that rhamnolipids have antibiotic activity, especially the double rhamnolipids can inhibit the growth of breast cancer cells (Bioresource Technology 2007, 98: 1149-1153), and help psoriasis, neurodermatitis, etc. Treatment of skin diseases (Journal of Dermatological Science 2005, 40: 141-143) and autoimmune diseases (U.S. Patent 5, 466, 675, 1999). These discoveries have la...

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): C12N1/00G01N30/90
Inventor 杨树林孙崇艾凤祥孟欣欣钮小松王永斌孟广荣王影
Owner NANJING UNIV OF SCI & 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

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