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Rhamnolipid efficient fermentation technology

A fermentation process, rhamnolipid technology, applied in the field of high-efficiency fermentation process of rhamnolipids, can solve the problems of low fermentation efficiency and high cost, achieve rapid growth and reproduction, reduce raw material costs, high market competitiveness and application potential Effect

Inactive Publication Date: 2018-07-31
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a high-efficiency fermentation process for rhamnolipids, so as to solve the problems of high cost and low fermentation efficiency in the prior art

Method used

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  • Rhamnolipid efficient fermentation technology
  • Rhamnolipid efficient fermentation technology
  • Rhamnolipid efficient fermentation technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: Pseudomonas KT1115 plate activation

[0039] (1) Prepare 50 mL of LB solid medium, place it in a 250 mL Erlenmeyer flask, sterilize by high-pressure steam at 121 °C for 20 min, and cool it naturally to about 60 °C after completion.

[0040] (2) Prepare a number of disposable petri dishes in an ultra-clean workbench, pour about 20mL of medium into each petri dish, cool and solidify for later use.

[0041] (3) Take out the seed preservation tube from -80°C, place it in a refrigerator at 4°C to melt slowly, then use a sterile inoculation loop to dip in the melted bacterial solution, and streak it on the LB agar plate.

[0042] (4) Plates are placed in a constant temperature incubator at 30°C, and cultured upside down for 12-16 hours, then a single colony is picked and transferred to an LB agar plate for secondary activation, and cultured upside down at 30°C for 6-8 hours until a clear single colony grows on the plate. colony.

Embodiment 2

[0043] Embodiment 2: Preparation of Pseudomonas KT1115 Fermented Seed Liquid

[0044] (1) Prepare 150 mL of LB liquid medium, put it in a 500 mL Erlenmeyer flask, sterilize it by high-pressure steam at 121°C for 20 minutes, and complete natural cooling.

[0045] (2) Preparation of seed solution: Pick a single colony from the secondary purification plate in Example 1 with an inoculation loop, inoculate it in LB liquid medium, place it in a constant temperature shaker at 30° C., and cultivate it at 200 rpm.

[0046] (3) After culturing for 20 hours, start sampling to detect its OD 600 Absorbance, cells at OD 600 The absorbance is 5.0 ~ 6.0 can be inoculated.

Embodiment 3

[0047] Example 3 Determination of Rhamnolipid Production Ability by Single Carbon Source Fermentation

[0048] The fermentation medium formula (excluding carbon source) is: yeast extract 3g / L, NaNO 3 6g / L, KH 2 PO 4 1g / L, Na 2 HPO 4 1g / L, CaCl 2 2H 2 O 0.11g / L, MgSO 4 0.1g / L.

[0049] The carbon sources of the fermentation medium are: glycerol 20g / L, soybean oil 20g / L, corn oil 20g / L, rapeseed oil 20g / L, sunflower oil.

[0050] The experiment adopts shake flask fermentation. Add 50mL of the prepared fermentation medium into a 250mL conical flask, and sterilize it with high-pressure steam at 121°C for 20 minutes; when it is naturally cooled to 30°C, take 3mL of the seed solution of Example 2 and put it into the conical flask, place it in a constant temperature oscillator at 30°C, and set The rotating speed is 200rpm, after 192 hours of fermentation, the measured rhamnolipid contents are: glycerol 10.8g / L, soybean oil 11.5g / L, corn oil 6.5g / L, rapeseed oil 15.7g / L, ...

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Abstract

The invention discloses a rhamnolipid efficient fermentation technology. A fermentation strain has good hereditary stability and can meet the production demands. After optimization of the fermentationcondition, the strain utilizes 30g / L of glycerol and 60g / L of rapeseed oil as a compound carbon source, and after fermentation for 192h in a 5L fermentation tank, the highest yield of rhamnolipid reaches 53.3g / L. Compared with the prior art, by adopting the rhamnolipid efficient fermentation technology disclosed by the invention, the rhamnolipid with an equal yield can be acquired under the condition of equal carbon source supply; and meanwhile, as the cost of industrial glycerol is low, the production cost is lowered by utilizing glycerol as a substrate, and thus the application potential ofthe rhamnolipid as a biosurfactant is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a high-efficiency fermentation process of rhamnolipids. Background technique [0002] Biosurfactants are derived from microorganisms, animals and plants, and are composed of hydrophilic groups (acids, cations, anions, monosaccharides, disaccharides or polysaccharides) and hydrophobic groups (saturated, unsaturated hydrocarbon chains or fatty acids). Compared with chemically synthesized surfactants, biosurfactants have more complex chemical structures and more functional groups. They have various surface active functions such as reducing liquid surface / interfacial tension, forming micelles, and two-phase microemulsions. Excellent thermal and chemical stability, and the product itself is non-toxic, environmentally friendly, can be degraded by environmental microorganisms, and has good biocompatibility. It is a typical environmentally friendly "green product". In addition, ...

Claims

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Application Information

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IPC IPC(8): C12P19/44C12R1/385
CPCC12P19/44
Inventor 董维亮徐宁信丰学周杰马江峰姜岷
Owner NANJING UNIV OF TECH
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