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Rhamnolipid high-yielding bacterial strain, and applications thereof

A rhamnolipid, high-yield technology, applied in the field of Pseudomonas aeruginosa, can solve the problems of restricting the application of rhamnolipid, high production cost, low yield, etc., achieve excellent surface activity, reduce production cost, and promote bacterial growth The effect of growth

Inactive Publication Date: 2017-07-28
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current low yield and high production cost limit the wide application of rhamnolipids in industry. Therefore, obtaining a high-yield strain and utilizing low-cost or waste raw materials is very important for the industrial production of rhamnolipids.

Method used

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  • Rhamnolipid high-yielding bacterial strain, and applications thereof
  • Rhamnolipid high-yielding bacterial strain, and applications thereof
  • Rhamnolipid high-yielding bacterial strain, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] This example illustrates the method of performing ARTP mutagenesis on the original strain of Pseudomonas aeruginosa, and performing primary screening and secondary screening to screen excellent Pseudomonas aeruginosa.

[0036] The original strain of Pseudomonas aeruginosa CICC 21100 was derived from CICC, the specific method is as follows:

[0037] Helium was used as the gas, the ventilation flow rate was 10 L / min, the action distance was 2 mm, and the action power was 120 W. Dilute the seeds of Pseudomonas aeruginosa CICC 21100 with 0.9% saline to OD 600 =0.6, take 10 μL of the bacterial solution and evenly spread it on a sterile small iron sheet, air-dry it and put it in a breeding machine for 120 s of mutagenesis to obtain positive mutants.

[0038] Wherein, the medium formula used (% is mass percentage):

[0039] Blue gel slab primary screening: Dextrose 2.0%, KH 2 PO 4 0.07%, Na 2 HPO 4 0.09%, NaNO 3 0.2%, MgSO 4 ·7H 2 O 0.04%, CaCl 2 2H 2O 0.01%, CTAB...

Embodiment 2

[0058] This example illustrates the passage stability and taxonomic characteristics of the mutant strain KT1115.

[0059] The mutant strain KT1115 was subcultured and fermented every 12 h, and the rhamnolipid production of each generation of fermentation liquid was detected to observe whether there was a back mutation. The results are shown in Table 2.

[0060] Table 2 The rhamnolipid production of strain KT1115 with different generations

[0061]

[0062] From the experimental results, it can be seen that after 7 consecutive passages, the amount of rhamnolipid produced by the mutant strain KT1115 is relatively stable, and it has good passage stability, which can be used as a production strain for further research and development.

[0063] Taxonomic characteristics of the mutant strain KT1115: the cell size is 0.5-0.8 μm × 1.5-3.0 μm, with a single flagella. Colonies are flat with irregular edges.

Embodiment 3

[0065] This example illustrates Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) strain KT1115 fermented to produce rhamnolipid with waste oil as the only carbon source.

[0066] Waste oil pretreatment: The waste oil collected from the canteen was first allowed to stand for 12 hours, and filtered with qualitative filter paper; then the filtrate was left to stand for 12 hours, and then filtered with qualitative filter paper, and the filtrate was obtained as waste oil for experiment.

[0067] The medium formula described in this embodiment (% is mass fraction):

[0068] Seed Medium: Peptone 1.0%, Yeast Extract 0.5%, Sodium Chloride 1.0%, pH 7.0.

[0069] Fermentation medium: waste oil 1.0%, NaNO 3 0.5%, KH 2 PO 4 0.1%, Na 2 HPO 4 12H 2 O 0.1%, MgSO 4 0.01%, FeSO 4 ·7H 2 O 0.02%, CaCl 2 2H 2 O 0.01%, pH6.0.

[0070] Inoculate 5 μL of Pseudomonas aeruginosa KT1115 stored at -80°C into 5 mL of LB seed medium, and culture overnight at 37°C and 200 rpm. The seed solu...

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Abstract

The invention discloses a rhamnolipid high-yielding bacterial strain, and applications thereof. The rhamnolipid high-yielding bacterial strain is named as pseudomonas aeruginosa KT1115, is preserved at China Center for Type Culture Collection, and the preservation number is CCTCC M 2016686. The rhamnolipid high-yielding bacterial strain is stable in hereditary features and high in rhamnolipid yield, and is obtained via ARTP induced mutation, blue gel plate screening, and secondary screening. According to fermentation broth of pseudomonas aeruginosa KT1115, liquid surface tension is reduced to 24.8mN / m<2>, and emulsification value is as high as 80%. Optimization of fermentation conditions including carbon source and nitrogen source single factors, cheap crude glycerine and swill-cooked dirty oil are taken as a composite carbon source, so that production cost is reduced greatly. The rhamnolipid yield of pseudomonas aeruginosa KT1115 is 50.7g / L, is increased by 18.1 times compared with that of a starting strain, conversion rate is 0.75g / g, and is relatively high. Application prospect of pseudomonas aeruginosa KT1115 in further industrialized application is promising.

Description

technical field [0001] The invention relates to the technical field of biological fermentation, in particular to a strain of Pseudomonas aeruginosa obtained through mutagenesis and selection, and the application of the Pseudomonas aeruginosa. Background technique [0002] Biosurfactant is a kind of surfactant synthesized by microorganisms with various structures, including glycolipids, lipopeptides, lipoproteins, phospholipids and neutral lipid derivatives. Surfactants generally consist of hydrophilic groups and hydrophobic groups. Rhamnolipid is an anionic biosurfactant belonging to glycolipids, which has good washing, emulsifying, solubilizing, wetting, penetrating and other properties, and has good biodegradability. It has great application prospects in the fields of bioenergy, petrochemical, biomedicine, food and daily chemicals. [0003] The hydrophilic groups of rhamnolipids generally consist of 1-2 molecules of rhamnose, and the hydrophobic groups consist of 1-2 mol...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P19/44C12R1/385
CPCC12P19/44C12R2001/385C12N1/205
Inventor 姜岷徐帆莉董维亮马江锋吴昊信丰学
Owner NANJING UNIV OF TECH
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