Method for preparing rhamnolipid

The technology of rhamnolipid and strain is applied in the field of preparation of rhamnolipid, which can solve the problems of bacterial contamination of culture medium, long screening period, difficulty in judging the production of surfactants, etc., and achieves an obvious effect of environmental remediation. , Improve the screening accuracy, the effect of significant emulsification

Inactive Publication Date: 2007-01-10
NANJING UNIV OF SCI & TECH
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  • Claims
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Problems solved by technology

There are following deficiencies in the above-mentioned method for preparing rhamnolipids: 1, adopt the dilution coating method of blood agar plate to carry out the screening of producing rhamnolipid bacterial strains, although the generation of hemolytic circle can be seen visually, but hemolytic circle is irregular, Some components or miscellaneous bacteria in the blood often produce similar hemolytic circles. In actual operation, it is difficult to judge the amount of surfactant produced by the size of the hemolytic circle, and during the culture process, some components or miscellaneous bacteria in t

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  • Method for preparing rhamnolipid

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Embodiment 1

[0016] Embodiment 1: in conjunction with accompanying drawing, the preparation method of rhamnolipid of the present invention, comprises the following steps:

[0017] 1.1 Bacteria screening: The samples from oilfield sewage or oil sludge were soaked and pretreated, and then inserted into the proliferation medium. The cultured culture solution was inoculated on a blood agar plate by streaking and cultured, and a single colony in the hemolysis zone was picked. And inoculated on the blue agar plate for culture, pick the colony with blue halo and insert it into the slant for strain preservation; insert the primary screening strain stored in the slant medium into the fermentation medium and measure its oil discharge after fermentation Activity, emulsifying performance and surface tension of fermentation broth, and then select the target strain and co-exist; that is, select samples from oilfield sewage or oil sludge, soak them in sterile water for 2 hours for pretreatment, take out t...

Embodiment 2

[0029] Example 2: The preparation method of the rhamnolipid of the present invention, sampling from oil pollution in the oil field, and screening through enrichment culture, blood plate (primary screening) and blue gel plate composite screening mode (rescreening) to obtain biosurfactants Produced bacteria, and improved by ultraviolet mutation breeding, the obtained excellent strains were preserved in the PDA slant. Culture by fermentation, i.e. fermentation medium: liquid paraffin or glucose or soybean oil or diesel oil or glycerol 15-35mL, (NH 4 ) 2 SO 4 2-2.5g, Na2HPO 4 ·2H 2 O3-3.8g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O 0.5g, CaCl 2 ·2H 2 O 0.005g, dilute to 1L with deionized water, and adjust pH to 7.5-8. Culture conditions: temperature: 26~36℃, shaker speed: 14~240r / min; volume of conical flask: 50-100mL; seed culture time: 21h; inoculation volume: 5~10%; culture time: 48~158h , the fermentation broth was extracted with chloroform and methanol, and the initial produc...

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Abstract

The invention discloses a manufacture method for rhamnolipid that includes the following steps: filtering fungus, inoculating into culture medium after taking dipping preprocess to the sample getting from oil field sewage or grease, inoculating on blood agar plate for culturing, selecting single colony, inoculating on blue agar plate for culturing, selecting the colony having blue halo to the bevel for microbial conserving; fermenting the primary screening strain in fermenting culture medium, testing the oil drainage activity, emulsibility and fermenting liquid surface tension, selecting target strain; taking ferment to the selected strain, and optimizing the fermenting culture medium, fermenting condition and strain to gain rhamnolipid fermenting liquid; distilling rhanmolipid raw product from the rhamnolipid fermenting liquid. The invention has high yield, good heavy hydrocarbon degradation effect, and improves the selecting accuracy.

Description

1. Technical field [0001] The invention discloses a preparation method of glycolipid biosurfactant, particularly a preparation method of rhamnolipid. 2. Background technology [0002] Rhamnolipid belongs to a glycolipid biosurfactant. In addition to reducing surface tension, stabilizing emulsion and increasing foam, it also has non-toxic and biodegradable properties that general chemical synthetic surfactants do not have. advantage. Therefore, it can be widely used in various fields of industry, agriculture, medicine and daily life, especially in the fields of petroleum industry and environmental engineering, such as biological viscosity reduction of petroleum, enhanced oil recovery, bioremediation of heavy oil polluted soil, etc. In addition, as a natural additive, rhamnolipid is more and more popular in the food industry, fine chemical industry, medicine and agriculture. [0003] In the prior art, there have been reports of synthesizing rhamnolipids by microbial methods....

Claims

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Application Information

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IPC IPC(8): C12P19/02
Inventor 杨树林宁长发沈薇陆晓唐仕荣孟广荣高力虎
Owner NANJING UNIV OF SCI & TECH
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