Industrial preparation method of rhamnolipid biological fermentation liquor

A rhamnolipid and bio-fermentation technology, which is applied in biochemical equipment and methods, microorganism-based methods, fermentation, etc., can solve the problems of no preparation method, high cost, and low efficiency in the industrialized large-scale production stage, and achieve a short cycle , improved recovery and high efficiency

Active Publication Date: 2008-05-14
BEIJING VICTEX ENVIRONMENTAL PROTECTION TECH DEV
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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

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  • Industrial preparation method of rhamnolipid biological fermentation liquor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Embodiment 1, chemical mutagenesis screening high-yield strains:

[0011] The medium used in the following experiments is as follows: Enrichment medium: 3g of beef extract, 10g of peptone, 5g of NaCl, 1000mL of double distilled water and 20g of agar, pH7.0~7.2. Blood agar plate medium: this medium was purchased from Beijing Yide Yihua Technology Co., Ltd.; blue agar plate medium: ammonium sulfate 0.1%, sodium nitrate 0.2%, magnesium sulfate 0.03%, potassium dihydrogen phosphate 1%, hydrogen phosphate Disodium 0.4%, yeast powder 0.05% and the rest of water, pH 7.0-7.2, coarse filter paper soaked in crude oil.

[0012] Screening steps:

[0013] 1. Enrichment culture of bacterial strains:

[0014] 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.

[0015] 2. Separation and purification:

[0016] Take 0.2...

Embodiment 2

[0044] Embodiment 2, industrialized method produces rhamnolipid biological fermentation liquid:

[0045] Shake flask and fermenter medium formulations are: corn oil 2%, urea 1%, molasses 15%, KCl 0.1%, KH 2 PO 4 0.05%, K 2 HPO 4 0.3%, yeast extract 0.005%, composite trace elements 0.008%, the rest is water, pH is 6.5, the composite trace elements are Zn 1.5g / L, Cu 1.0g / L, Mn 1.0g / L, Se 1.5g / L, Co 1.0g / L and solvent water.

[0046] Shake flask and fermenter culture:

[0047] 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-t...

Embodiment 3

[0048] Embodiment 3, industrialized method produces rhamnolipid biological fermentation liquid:

[0049] Shake flask and fermenter medium formulations are: corn oil 3%, urea 1.5%, molasses 17%, KCl 1.0%, KH 2 PO 4 0.2%, K 2 HPO 4 1.5%, yeast extract 0.02%, composite trace elements 0.01%, the rest is water, pH is 6.5, the composite trace elements are Zn 1.5g / L, Cu 1.0g / L, Mn 1.0g / L, Se 1.5g / L, Co 1.0g / L and solvent water.

[0050] Shake flask and fermenter culture:

[0051] 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 ve...

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Abstract

The invention relates to a preparation method of rhamnolipid biological fermentation liquid. The method comprises the following steps: extracting and purifying Pseudomonas aeruginosa obtained from petroleum-contaminated soil, and obtaining Pseudomonas aeruginosa VTS-1 (Pseudomonas sp. -1) The preservation number is: CGMCC2200, and the specific fermentation and cultivation steps are as follows: A. Strain VTS-1 is cultured in shake flasks; B. Primary fermentation cultivation; C. Secondary fermentation cultivation; D. Fermentation cultivation. The process of the method has the advantages of high yield, high efficiency, high loading coefficient, short fermentation cycle, simple process, reduced overall product cost, and is completely suitable for industrial production, and has played a role in the promotion and large-scale application of rhamnolipid biosurfactants. important role.

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 conten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/02C12R1/385
Inventor 李玉梅金艳芳马艳玲沈超沈玉江康婷婷王芳李伟徐远洋陈韶军
Owner BEIJING VICTEX ENVIRONMENTAL PROTECTION TECH DEV
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