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Antifouling composition prepared from a pseudomonas pf-11 culture

一种PF-11、假单胞菌的技术，应用在植物学设备和方法、生物化学设备和方法、细菌等方向，能够解决大代谢差异寄生虫、宿主不同传播和感染策略、困难等问题

Inactive Publication Date: 2018-06-08
BIOMIMETX SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Several factors explain the difficulties experienced when known antiparasitic compounds have been tested on new species, such as greater genotypic and phenotypic diversity among different species of parasites, greater metabolic Differences and the fact that parasites occupy very different habitats and have different transmission and infection strategies towards hosts

Method used

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  • Antifouling composition prepared from a pseudomonas pf-11 culture
  • Antifouling composition prepared from a pseudomonas pf-11 culture
  • Antifouling composition prepared from a pseudomonas pf-11 culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0165] Example 1 - Isolation, Characterization and Storage of Strain PF-11

[0166] Environmental Sampling and Bacterial Isolation

[0167] Soil and / or mud samples were collected in the Tagus River region around Lisbon, Portugal. The collected material (10 g) was homogenized with sterile water (50 mL). After gravity settling of the mixture, the liquid fraction was recovered. Then, the raw material suspension (including microorganisms) was collected by centrifugation (12000 g, 5 min). The resulting pellet was resuspended in sterile water. Primary growth was accomplished in LB (Luria Bertani) medium. These cultures were diluted (10-2 to 10-9) and grown on LA (LB+agar) or with ampicillin (8 μg / mL), amoxicillin (8 μg / mL) or cefotaxime (cefotaxime) (2 μg / mL) was grown in LA to select for resistant or reduced susceptibility strains. Colonies with significant size or morphology differences were selected. Each selected colony was subjected to successive plate passages (up to ...

Embodiment 2

[0174] Example 2 - PF-11 Culture Growth, Compound Preparation, Recovery and Characterization of Secreted Compounds

[0175] PF-11 growth conditions.

[0176]Frozen bacterial aliquots were grown overnight (16-18 h) at 30°C in LB agar medium. One colony was then used to transfer into a sterile flask containing M9 medium supplemented with glucose and grown in a bacterial constant temperature shaker at 120 r.p.m. for 16 h at 30°C.

[0177] Compounds are recovered from the culture.

[0178] Cells were removed by centrifugation (14.000 r.p.m., 4°C, 15 min) and the supernatant was collected. The supernatant was sterilized by filtration using a filter unit with a 0.22 μm DURAPORE filter (Millex GP, Millipore, Ireland). Sterility was verified by incubating 50 μl of the supernatant in LA dishes at 30° C. for at least 16 h.

[0179] Purification of original mixtures of compounds.

[0180] Supernatants were frozen at -80°C and dehydrated by lyophilization. It was then resuspended ...

Embodiment 3

[0187] Example 3 - Antimicrobials from Environmental Sources

[0188] During a previous study, a heterogeneous collection of environmental Pseudomonas putida strains with strong adaptive skills collected through antibiotic resistance (Meireles 2013) was used to screen for secreted natural compounds Potential for microbial growth control. The content of Meireles 2013 is hereby incorporated by reference into this application. A set of P. putida isolates from this collection was selected based on its fitness level, antibiotic resistance and general fitness (data not shown), and aimed to collect multiple strain characteristics . . The secretome of these strains (ie the molecules they secrete) were collected and tested for the first time for effects on the growth of three model strain species E. coli, Staphylococcus aureus and Pseudomonas aeruginosa. One strain, PF-11, showed significant antibacterial potential. This initial set was then extended to all P. putida strains fro...

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Abstract

This invention concerns a method for preparing a bacterial supernatant comprising culturing a cell of Pseudomonas environmental strain PF-11; and recovering the supernatant. This invention also concerns a method for reducing the amount of a biofilm on a surface, reducing adhesion of at least one organism to a surface, or reducing microfouling or macrofouling on a surface comprising contacting thesurface with a supernatant, supernatant fraction, modified supernatant or modified supernatant fraction of Pseudomonas strain PF-11; or a composition comprising a supernatant, supernatant fraction, modified supernatant or modified supernatant fraction of Pseudomonas strain PF-11, and one or more acceptable carriers. This invention also concerns a method for killing or reducing the growth of a fungus or bacterial cell, or killing or inhibiting the development of an insect or marine copepod, comprising contacting the fungus, bacteria, insect or marine copepod with a supernatant, supernatant fraction, modified supernatant or modified supernatant fraction of a Pseudomonas strain PF-11 culture; or a composition comprising a supernatant, supernatant fraction, modified supernatant or modified supernatant fraction of a Pseudomonas strain PF-11 culture, and one or more acceptable carriers. This invention also concerns a substantially pure culture of Pseudomonas strain PF-11. This invention alsoconcerns a culture that is enriched in Pseudomonas strain PF-11. This invention also provides a method of identifying whether a bacterium is capable of producing one or more extracellular proteases capable of digesting a high molecular weight substrate.

Description

[0001] Throughout this application, various publications are cited, including those within parentheses. Full citation information for publications cited in parentheses can be found at the end of the specification preceding the claims. The disclosures of all cited publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains. Background technique [0002] Antimicrobial Resistance [0003] Treating and preventing bacterial infections is a major health challenge worldwide over the next decade due to increasing bacterial resistance to antimicrobial agents (Kaplan 2004). Due to multidrug-resistant bacteria, more than 25,000 patients die of infection in the European Union alone each year, causing a total direct cost to society of 1.5 billion € (ECDC / EMA joint technical report 2009). But antibiotics used to treat human pathogens are also used in animals to treat dise...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/02A01N63/02A01N63/27
CPCC12N9/52C12N1/20C12Q1/04A01N63/27C12N1/02C12N9/485C12Y304/00A01P1/00A01P3/00C12R2001/38
Inventor 冈萨洛·科斯塔帕特里克·弗莱雷罗曼娜·桑托斯安娜·克里斯蒂娜·席尔瓦伊内斯·吉诺特
Owner BIOMIMETX SA
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