Pseudomonas aeruginosa and application thereof

A technology of Pseudomonas aeruginosa and rhamnolipid, which is applied in the directions of bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problems of easy contamination of miscellaneous bacteria, difficult to control fermentation process, loss of medium nutrients and bacterial cells, etc. , to achieve the effect of high proportion of disglycolipids, reducing the probability of contamination, and enriching the variety of substrates

Active Publication Date: 2018-01-09
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Biosurfactants have a good foaming effect, so measures such as stirring and aeration during the fermentation process will cause the fermentation liquid to overflow in the form of foam, which not only loses the nutrients and bacteria of the medium, but also easily contaminates miscellaneous bacteria
Especially in the seed culture process, the rhamnolipids accumulated during the seed culture process lead to a large amount of foam overflow, which can easily lead to bacterial contamination in subsequent fermentation
At present, the main factors that limit the wide application of rhamnolipids in industry are its high production cost and complicated and difficult-to-control fermentation process.

Method used

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  • Pseudomonas aeruginosa and application thereof
  • Pseudomonas aeruginosa and application thereof
  • Pseudomonas aeruginosa and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] This example illustrates the method for obtaining the strain Pseudomonas aeruginosa YM4 of the present invention.

[0028] The original strain of the bacterium was obtained from the soil around Shengli Oilfield and inoculated into LB liquid medium, and cultivated at 37°C and 200rpm for 12h to obtain a seed solution. Use a pipette gun to take 100 μL of the seed liquid in a sterile test tube of No. 1 900 μL sterile water in the ultra-clean workbench, shake it fully, and disperse the bacterial liquid evenly, that is, 10 -1 sample diluent; take another 10 -1 100 μL of sample diluent in No. 2 sterile test tube of 900 μL sterile water, and so on, mark 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 and 10 -8 . Take 10 respectively -7 and 10 -8 Dilution 200 μL of the dilution solution is evenly coated on the plate with a sterile coating rod, and it is placed flat for 10 minutes, and then it is inverted and cultured in a 37°C constant temperature incubator for 18-24 hours to...

Embodiment 2

[0034] This example illustrates the method for Pseudomonas aeruginosa YM4 provided by the present invention to use glycerol as a carbon source to produce high biosurfactant rhamnolipid.

[0035] Seed medium (LB medium): peptone 10.0g / L, yeast powder 5.0g / L, NaCl 10.0g / L.

[0036] Initial fermentation medium: glycerol 40.0g / L, NaNO 3 2.5g / L, K2 HPO 4 ·3H 2 O 4.0g / L, KH 2 PO 4 4.0g / L, NaCl 1.0g / L, KCl 1.0g / L, MgSO 4 ·7H 2 O 0.2g / L, anhydrous CaCl 2 0.1g / L, yeast powder 1.0g / L, pH 6.5.

[0037] Optimized fermentation medium: glycerol 30.0~60.0g / L, NaNO 3 2.0~12.0g / L,K 2 HPO 4 ·3H 2 O 4.0g / L, KH 2 PO 4 4.0g / L, NaCl 1.0g / L, KCl 1.0g / L, MgSO 4 ·7H 2 O 0.2g / L, anhydrous CaCl 2 0.1g / L, yeast powder 1.0g / L, trace elements 2mL (FeCl 3 0.16g / L, CuSO 4 0.15g / L, ZnSO 4 ·7H 2 O 1.5g / L, MnSO 4 ·H 2 O1.5g / L), pH 7.

[0038] Pseudomonas aeruginosa YM4 provided by the present invention is first activated bacterial classification at 37 ℃ plate, after 20h, a ring of ...

Embodiment 3

[0042] This example illustrates the method for Pseudomonas aeruginosa YM4 to use crude glycerol as a carbon source to high-yield rhamnolipids.

[0043] Seed medium (LB medium): peptone 10.0g / L, yeast powder 5.0g / L, NaCl 10.0g / L.

[0044] Fermentation medium: glycerol 30.0g / L, NaNO 3 12.0g / L,K 2 HPO 4 ·3H 2 O 4.0g / L, KH 2 PO 4 4.0g / L, NaCl 1.0g / L, KCl 1.0g / L, MgSO 4 ·7H 2 O 0.2g / L, anhydrous CaCl 2 0.1g / L, yeast powder 1.0g / L, trace elements 2mL (FeCl 3 0.16g / L, CuSO 4 0.15g / L, ZnSO 4 ·7H 2 O 1.5g / L, MnSO 4 ·H 2 O 1.5g / L), pH 7.

[0045] The content of crude glycerol is 80% (w / w), and 37.5 g of crude glycerol is used per liter of fermentation broth.

[0046] Pseudomonas aeruginosa YM4 provided by the present invention is at first 37 DEG C plate activated bacterial classification, after 20h, connect a ring of well-grown thalli into 50mL LB culture medium, 37 DEG C, 200rpm cultivates 24h, then according to 3% (v / v ) transfer the amount of inoculum into 50mL f...

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Abstract

The invention relates to pseudomonas aeruginosa and an application thereof. The pseudomonas aeruginosa is named as Pseudomonas aeruginosa YM4 according to bacterial strain classification, and has a preservation number of CCTCC NO:M 2017494. The bacterial strain can produce rhamnolipid in a high yield from hydrophilic carbon sources, hydrophilic carbon sources and composite carbon sources, and therhamnolipid has a substrate conversion rate reach 65-80%. In a rhamnolipid product of the YM4 bacterial strain, di-rhamnolipid and single rhamnolipid are main products, and the amount of di-rhamnolipid and single rhamnolipid takes 80-90% of the total amount of glycolipid. According to the carbon source metabolic characteristic of the bacterial strain, a foam-free seed medium formula suitable for industrial application is developed, and the contamination rate of a seed tank is greatly reduced. The bacterial strain is high in rhamnolipid yield, conversion rate and ratio of a diglycolipid component, and has high industrial application value.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a pseudomonas aeruginosa strain for synthesizing rhamnolipids and a biosurfactant produced by the bacteria. Background technique [0002] Rhamnolipid is a biosurfactant that has been studied extensively in recent years. It not only has the properties of solubilization, emulsification, wetting and foaming comparable to chemical surfactants, but also has low toxicity, easy biodegradation, The characteristics of environmental friendliness make it a potential substitute for synthetic surfactants, so it has great application potential in fields such as petroleum exploration, medicine, food, daily chemicals, and environmental protection. Rhamnolipids accumulated by microbial metabolism are usually a mixture of a series of compounds composed of rhamnose in the hydrophilic group and long-chain β-hydroxy fatty acids in the hydrophobic group, including monorhamnolipids and dirhamnolipi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/44C12R1/385
Inventor 李霜张玉珉王丹范艳秋
Owner NANJING UNIV OF TECH
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