CN107595822A - Farnesol is preparing the application in suppressing the generation of pseudomonas aeruginosa pyo and biofilm formation medicine - Google Patents
Farnesol is preparing the application in suppressing the generation of pseudomonas aeruginosa pyo and biofilm formation medicine Download PDFInfo
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- CN107595822A CN107595822A CN201710737553.9A CN201710737553A CN107595822A CN 107595822 A CN107595822 A CN 107595822A CN 201710737553 A CN201710737553 A CN 201710737553A CN 107595822 A CN107595822 A CN 107595822A
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- farnesol
- pseudomonas aeruginosa
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Abstract
The invention discloses farnesol to prepare the application in suppressing the generation of pseudomonas aeruginosa pyo and biofilm formation medicine.Present invention discover that growth of the farnesol to pseudomonas aeruginosa PAO1 will not produce obvious inhibitory action, but the generation of pseudomonas aeruginosa PAO1 pyo can be significantly inhibited, while suppress the formation of its biomembrane.Therefore farnesol can be used for preparing the medicine for suppressing pseudomonas aeruginosa production pyo and suppress the medicine that aeruginosa biofilm is formed.
Description
Technical field
The invention belongs to microbial technology field, and in particular to farnesol is preparing suppression pseudomonas aeruginosa pyo
Produce and the application in biofilm formation medicine.
Background technology：
Pseudomonas aeruginosa (Pseudomonas aeruginosa) is also known as Pseudomonas aeruginosa, extensive in distributed in nature, is
One of common bacterium in soil.Pseudomonas aeruginosa is the conditioned pathogen of the mankind, is a series of serious pyogenic infections,
The especially important pathogenic bacteria of the underlying diseases scabies secondary infection such as bronchiectasis, chronic bronchitis, capsule pulmonary fibrosis.With
The extensive use of antibiotic, the Antibiotic Resistance of microorganism is increasingly wider, resistance intensity more and more higher, and the speed that drug resistance is formed also is got over
Come faster.Antibiotic it is lasting using providing lasting selection pressure for high antibody-resistant bacterium, and then accelerate super resistance with
The formation of multiple antibiotic resistant strain.Pseudomonas aeruginosa can be to the quick generation drug resistance of Multiple Classes of Antibiotics.Human needs, which changes, causes a disease
The Management strategy of bacterium, find the new action target different from antibiotic.
Farnesol is a kind of 15- carbon sesquiterpene alcohols, and it is widely present in natural plants, such as gumbo seed, lemongrass, lemon
Lemon grass, peach, strawberry, tomato and corn etc..Candida albicans can also produce farnesol, and it is a kind of signal of candida albicans
Molecule.
The content of the invention：
First purpose of the present invention is to provide a kind of farnesol in control pseudomonas aeruginosa production pyo medicine
Application.
Make experiments show that growth of the farnesol to pseudomonas aeruginosa PAO1 will not produce obvious suppression
With, but the generation of pseudomonas aeruginosa PAO1 pyo can be significantly inhibited, while the shape of its biomembrane can be suppressed
Into.
Therefore, the present invention provides application of the farnesol in the medicine for suppressing pseudomonas aeruginosa production pyo is prepared.
Second object of the present invention is to provide farnesol and is preparing the medicine of suppression aeruginosa biofilm formation
In application.
Present invention discover that growth of the farnesol to pseudomonas aeruginosa PAO1 will not produce obvious inhibitory action, but can
The generation of pseudomonas aeruginosa PAO1 pyo is enough significantly inhibited, while the formation of its biomembrane can be suppressed, therefore can
Suppress the medicine that the medicine of pseudomonas aeruginosa production pyo and control aeruginosa biofilm are formed for preparing.
Brief description of the drawings：
Fig. 1 is the lower pseudomonas aeruginosa PAO1 of farnesol effect growth curve；
Fig. 2 is that farnesol suppresses kinetic curve caused by pseudomonas aeruginosa PAO1 pyo；
Fig. 3 is the block diagram for the biofilm formation that farnesol suppresses pseudomonas aeruginosa PAO1.
Embodiment：
Following examples are to further explanation of the invention, rather than limitation of the present invention.
Embodiment 1：
The preparation of PAO1 bacterium [pseudomonas aeruginosa (Pseudomonas aeruginosa)] suspension：By exponential phase of growth
Pseudomonas aeruginosa PAO1 nutrient solutions sample, and centrifugation, washed once, are resuspended in PBS with PBS, and bacteria concentration is dilute
Release to 108CFU/mL。
1. the influence experiment that farnesol grows to pseudomonas aeruginosa PAO1
LB culture mediums, farnesol are added into 5 test tubes respectively, and accesses the PAO1 bacteria suspensions of exponential phase of growth, it is overall
Product is 10mL, and the bacteria concentration for making pseudomonas aeruginosa PAO1 is 106CFU/mL, the concentration of farnesol is respectively 0 μ l/mL
(control), 0.16 μ l/mL, 0.31 μ l/mL, 0.63 μ l/mL and 1.25 μ l/mL.By several experimental groups it is separately sampled be added to from
In the special honeycomb culture plate of dynamic growth curve analyzer (Bioscreen C), 350 μ L nutrient solution, Mei Geshi are added per hole
It is parallel to test group three.Honeycomb culture plate is placed in automatic growth curve tester, 37 DEG C of shaken cultivations 3 days, determined per hour
OD600.With OD600For ordinate, incubation time is abscissa, the lower PAO1 of farnesol effect growth curve is drawn, with this
The influence that research farnesol grows to pseudomonas aeruginosa PAO1.As a result it is as shown in Figure 1.
2. farnesol caused by pseudomonas aeruginosa PAO1 pyos on influenceing to test
3 conical flasks, every bottle of access LB culture mediums, farnesol are taken, and accesses the PAO1 bacteria suspensions of exponential phase of growth, always
Volume is 100mL, and pseudomonas aeruginosa PAO1 bacteria concentration is 106CFU/mL, the concentration of farnesol is respectively 0 μ l/mL
(control), 0.16 μ l/mL and 0.63 μ l/mL.It is placed in shaking table 37 DEG C of constant-temperature shaking cultures 7 days, the green pus bacterium of sampling extraction daily
Element, every group of two Duplicate Samples.Extracting method is as follows：Nutrient solution 5mL is taken, centrifuges, supernatant is transferred to new pipe；3mL chloroforms are added, it is acute
Strong vibration, extract 30min；Centrifugation, chloroform layer is transferred in new pipe；1mL 0.2N hydrochloric acid is added, acutely vibration extracting 30min；
Centrifugation, upper strata aqueous phase is transferred in new pipe；In on ultraviolet specrophotometer at 520nm wavelength determine light absorption value.Using light absorption value as
Ordinate, incubation time are abscissa, draw the dynamics of the lower pseudomonas aeruginosa PAO1 pyo yield of farnesol effect
Change, farnesol is studied to influence caused by pseudomonas aeruginosa PAO1 pyos with this.Experiment is repeated twice.Experimental result
As shown in Figure 3.
3. farnesol caused by pseudomonas aeruginosa PAO1 biomembranes on influenceing to test
4 conical flasks, every bottle of access LB culture mediums, farnesol are taken, and accesses the PAO1 bacteria suspensions of exponential phase of growth, always
Volume is 10mL, and pseudomonas aeruginosa PAO1 bacteria concentration is 106CFU/mL, the concentration of farnesol is respectively 0 μ l/mL (right
According to), 0.16 μ l/mL, 0.63 μ l/mL and 1.25 μ l/mL.200 μ l are respectively taken to be added in No. 1 96- orifice plate from each experimental group,
Each experimental group respectively takes 5 Duplicate Samples.No. 1 96- orifice plate after sample-adding is placed in 37 DEG C of quiescent culture 24h, then by No. 1 96-
Nutrient solution in orifice plate is correspondingly transferred in one piece of No. 2 new 96- orifice plate.No. 1 96- orifice plate is washed with deionized and removed three times
Suspension cell, 40 DEG C of dry 2min in baking oven are subsequently placed in, 0.1% crystal violet for adding 300 μ l dyes to biomembrane
10min, decrystallize purple dye liquor, and the crystal violet for removing residual is washed with deionized.No. 1 96- orifice plate is placed in 40 in baking oven
DEG C dry 2min, then per hole add 300 μ l 95% ethanol extraction biomembrane in crystal violet 10min, then ethanol is extracted
Liquid is taken correspondingly to be transferred in one piece of No. 3 new 96- orifice plate.No. 2 96- orifice plates are placed in microplate spectrophotometer (Thermo
Scientific Multiskan GO) on measure OD600To obtain the relative concentration values of planktonic cells, No. 3 96- orifice plates are placed in
Should be to measure OD in orifice plate spectrophotometer590To obtain the fractional yield value of biomembrane.Draw the phase of planktonic cells and biomembrane
To the block diagram of yield.
Experimental result：
It will be seen from figure 1 that the farnesol treatment group of various concentrations and the growth of pseudomonas aeruginosa PAO1 in control group
Curve does not have significant difference, presents typical growth curve, including lag phase, exponential phase of growth, stationary phase and decline
Phase, because the growth curve is the OD with bacterium solution600Be worth for ordinate, reflection be bacterium living and dead bacterium total light absorption value, therefore
Decline phase unobvious.
Figure it is seen that farnesol inhibits the generation of pseudomonas aeruginosa PAO1 pyo, it is green in control group
The yield of pyocin gradually rose with the extension of incubation time before this, had reached maximum when cultivating 3 days, afterwards with training
The extension of time is supported, pyo yield is again on a declining curve.But in 0.16 μ l/mL and 0.63 μ l/mL farnesol treatment group,
The yield of pyo is significantly lower than control group always.Test result indicates that the farnesol of low concentration inhibits P. aeruginosa
The generation of the pyo of bacterium.
From figure 3, it can be seen that farnesol can suppress the formation of pseudomonas aeruginosa PAO1 biomembrane, control group and place
The increment of planktonic cells in reason group does not have significant difference, but in 0.16,0.63 and 1.25 μ l/mL farnesol treatment group
The yield of biomembrane is significantly lower than control group.Test result indicates that the farnesol of low concentration can suppress pseudomonas aeruginosa
The yield of biomembrane.
To sum up test result indicates that, the farnesol of low concentration will not suppress pseudomonas aeruginosa PAO1 growth (Fig. 1),
But the generation (Fig. 2) of pseudomonas aeruginosa PAO1 pyo can be suppressed, while the formation (figure of its biomembrane can be suppressed
3)。
Claims (2)
1. application of the farnesol in the medicine for suppressing pseudomonas aeruginosa production pyo is prepared.
2. application of the farnesol in the medicine for suppressing aeruginosa biofilm formation is prepared.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710737553.9A CN107595822A (en) | 2017-08-24 | 2017-08-24 | Farnesol is preparing the application in suppressing the generation of pseudomonas aeruginosa pyo and biofilm formation medicine |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710737553.9A CN107595822A (en) | 2017-08-24 | 2017-08-24 | Farnesol is preparing the application in suppressing the generation of pseudomonas aeruginosa pyo and biofilm formation medicine |
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| CN107595822A true CN107595822A (en) | 2018-01-19 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110063948A (en) * | 2019-05-14 | 2019-07-30 | 福建师范大学 | P-hydroxyphenylethanol is used to prepare the purposes for inhibiting the drug of bacterial community induction |
| CN113430158A (en) * | 2021-06-21 | 2021-09-24 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Preparation of geraniol in promoting pseudomonas aeruginosa 3OC12Application of HSL signal molecule synthesis preparation |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006290872A (en) * | 2005-03-15 | 2006-10-26 | Hirosaki Univ | Functional isoprenoid assuming anti-microbial activity against gram-negative bacterium and gram-positive bacterium or the like |
-
2017
- 2017-08-24 CN CN201710737553.9A patent/CN107595822A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006290872A (en) * | 2005-03-15 | 2006-10-26 | Hirosaki Univ | Functional isoprenoid assuming anti-microbial activity against gram-negative bacterium and gram-positive bacterium or the like |
Non-Patent Citations (3)
| Title |
|---|
| CARLA CUGINI 等: "Farnesol, a common sesquiterpene, inhibits PQS production in Pseudomonas aeruginosa", 《MOLECULAR MICROBIOLOGY》 * |
| H. M. H. N. BANDARA 等: "Incorporation of Farnesol Significantly Increases the Efficacy of Liposomal Ciprofloxacin against Pseudomonas aeruginosa Biofilms in Vitro", 《MOLECULAR PHARMACEUTICS》 * |
| HAN-SHIN KIM 等: "Raffinose, a plant galactoside,inhibits Pseudomonas aeruginosa biofilm formation via binding to LecA and decreasing cellular cyclic diguanylate levels", 《SCIENTIFIC REPORTS》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110063948A (en) * | 2019-05-14 | 2019-07-30 | 福建师范大学 | P-hydroxyphenylethanol is used to prepare the purposes for inhibiting the drug of bacterial community induction |
| CN113430158A (en) * | 2021-06-21 | 2021-09-24 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Preparation of geraniol in promoting pseudomonas aeruginosa 3OC12Application of HSL signal molecule synthesis preparation |
| CN113430158B (en) * | 2021-06-21 | 2022-05-10 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Preparation of geraniol in promotion of pseudomonas aeruginosa 3OC12Application of HSL signal molecule synthesis preparation |
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Application publication date: 20180119 |