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Industrial preparation method of rhamnolipid biological fermentation liquor

A rhamnolipid, biological fermentation technology, applied in biochemical equipment and methods, microorganism-based methods, fermentation, etc., to achieve the effects of high efficiency, short cycle, and improved recovery rate

Inactive Publication Date: 2011-06-08
BEIJING VICTEX ENVIRONMENTAL PROTECTION TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the production of rhamnolipids mostly adopts microbial fermentation, but it is limited to the laboratory stage. It is still in the stage of industrialized large-scale production and there is no matching preparation method, and the existing preparation methods are low in efficiency, high in cost and low in content. low, so applications are limited

Method used

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  • Industrial preparation method of rhamnolipid biological fermentation liquor
  • Industrial preparation method of rhamnolipid biological fermentation liquor
  • Industrial preparation method of rhamnolipid biological fermentation liquor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Screening steps:

[0014] 1. Enrichment culture of bacterial strains:

[0015] Weigh 1g of soil contaminated by crude oil for a long time and dissolve it in 100ml of sterile water, draw 1mL into the enrichment medium, and place it in a constant temperature shaking incubator at 37°C for 24 hours.

[0016] 2. Separation and purification:

[0017] Take 0.2ml of the bacterial suspension obtained after the enrichment culture and spread it on the blood agar plate, culture it in a constant temperature incubator at 37°C for 48 hours, select the strain with a large hemolytic circle around the colony and culture it in a shaker flask, and use the same method Separation, purification and culture for 48 hours after coating on blue plate.

[0018] 3. Mutagenesis improvement:

[0019] After the bacterial strains screened by the above method are made into a bacterial suspension and spread on the plate, put a little nitrosoguanidine crystal on the inner edge of the plate, and incubat...

Embodiment 2

[0047] Shake flask and fermenter culture:

[0048] Utilize the slant bacterial classification of embodiment 1 gained, cultivate 30h, transfer into 5L shaking flask (fill 2L substratum), inoculum size 5% (bacteria solution concentration is in OD 600 =0.8~1.0), the temperature is 32±2°C, 120rpm cultivated for 14h, transferred to a 500-liter primary fermenter, the filling coefficient is 70%, the inoculum size is 5%, the temperature is 32±2°C, and the ventilation ratio is 1:1, pH 6.5-7.5, stirring speed 100rpm, culture for 12 hours, transfer to a 3.5-ton secondary fermentation tank, the filling coefficient is 70%, the inoculum size is 5%, the temperature is 32±2°C, and the ventilation ratio is 1 : 1. The pH is 6.5-7.5, the stirring speed is 100rpm and cultivated for 12 hours, then transferred to a 30-ton three-stage fermenter with a loading capacity of 20 tons. The process parameters of the three-stage fermenter are: temperature 32±2°C, sterilization The pH is adjusted to about 6...

Embodiment 3

[0051] Shake flask and fermenter culture:

[0052] Utilize the slant bacterial classification of embodiment 1 gained, cultivate 30h, transfer into 5L shaking flask (filling into 2L culture medium), inoculum size 5%, 32 ± 2 ℃, 120rpm cultivate 16h, transfer into 500 liters of primary fermentation tank, the filling coefficient is 67%, the inoculum size is 5%, the ventilation ratio is 1:1 at 32±2°C, the pH is 6.5-7.5, the stirring speed is 120rpm, and the stirring speed is 120rpm. The material coefficient is 67%, the inoculum size is 5%, 32±2°C, the ventilation ratio is 1:1, the pH is 6.5~7.5, the stirring speed is 120rpm and cultivated for 12h, then transferred to a 30-ton three-stage fermenter, and the charging capacity is 20 tons, the process parameters of the three-stage fermentation tank are: temperature 32±2°C, pH adjusted to about 6 before sterilization, and 0.210 tons of NaOH to control the pH above 6 in the later stage, the ventilation ratio is 1:1, and the stirring spee...

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Abstract

The invention relates to a preparation method for rhamnolipid bio-fermentation fluid. The preparation method comprises the steps that: aeruginosa pseudomonas is produced by materials extracted from the soil which is contaminated by petroleum, separated and purified; the aeruginosa pseudomonas is mutated by nitroso guanidine, so as to obtain the aeruginosa pseudomonas VTS-1(Pseudomonas sp.VTS-1) with the preservation number: CGMCC2200. The fermentation culture has the steps that: A. the stain VTS-1 is inoculated into a shake flask to be cultured; B. a first grade fermenting culture is done; C.a second grade fermenting culture is done; D. the fermenting culture is done. The technique has the advantages of high yielding, high efficiency and charging coefficient, short fermentation period and simple technique, leading the product comprehensive cost to be lowered and being fully suitable for industrialized production and having great contribution to popularization and large-scale application of rhamnolipid bio surfactant.

Description

Technical field: [0001] The invention relates to a production method of a biosurfactant, in particular to an industrialized preparation method of a rhamnolipid biofermentation liquid. Background technique: [0002] Rhamnolipid belongs to a kind of glycolipid surfactant, which is an extracellular metabolite produced by microorganisms growing to a certain stage in the fermentation process under suitable conditions. It is a kind of biosurfactant with better effect. Like other synthetic surfactants, its molecular structure has both hydrophilic and hydrophobic properties, and can reduce surface tension. At the same time, it has its own characteristics: high biodegradability and lower biotoxicity, Low critical micelle concentration and higher surface activity, gradual absorption, continuous activity and other characteristics. At present, the research on rhamnolipids has always been in the stage of indoor research, and no industrial production scale has been formed, and the conte...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/02C12R1/385
Inventor 陈韶军金艳芳张生赵力隋志强汪成章于洋王守忠沈超刘志梅
Owner BEIJING VICTEX ENVIRONMENTAL PROTECTION TECH DEV
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