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Method for accessing microbial diversity

a technology of microbial diversity and access method, which is applied in the field of accessing microbial diversity, can solve problems such as difficulties experienced by microbiologists

Inactive Publication Date: 2011-03-17
FRAUNHOFER USA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Such a mechanism of sustaining a relatively low cell density may also contribute to the difficulties experienced by microbiologists in trying to establish pure cultures of these bacteria.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0036]Growth and isolation of novel microorganisms: Utilization of esterase Sigma E0887 to degrade homoserine lactone (Type 1) autoinducer signals.

[0037]In this example water from a lake / reservoir is used as the source of microorganisms, however any source that is expected to exhibit a diversity of novel (i.e., not yet cultured) microorganisms may be used.

[0038]Water samples (approximately 1-2 liters) are obtained at a depth of 0-2 meters from Marsh Creek Lake (Downingtown, Pa.) and Spruce Run Reservoir (Clinton, N.J.); the samples are maintained at 4-20 C until used.

[0039]Agar media are prepared consisting of 1) filter-sterilized sample water, 0.5% Difco Agar Noble (Becton Dickinson, Sparks, Md.), and 0.05% Difco Bacto Peptone and 2) filter-sterilized sample water, 0.5% Difco Agar Noble, and 0.50% Difco Bacto Peptone. Preferably the agar and peptone are prepared at 4× concentration in sample water, sterilized and then diluted with filter-sterilized sample water to the appropriate v...

example 2

[0045]Growth and isolation of novel microorganisms: Utilization of L-selenomethionine or adenine to inhibit the Type 2 autoinducer signal.

[0046]In this example water from the Atlantic Ocean is used as the source of microorganisms, however any source that is expected to exhibit a diversity of novel (i.e., not yet cultured) microorganisms may be used.

[0047]Water samples (approximately 1-2 liters) are obtained at a depth of 0-2 meters from the Atlantic Ocean, 1 mile east of Ocean City, Md.; the samples are maintained at 4 C until used.

[0048]Agar media are prepared consisting of 1) filter-sterilized sample water, 0.5% Difco Agar Noble (Becton Dickinson, Sparks, Md.), and 0.05% Difco Bacto Peptone and 2) filter-sterilized sample water, 0.5% Difco Agar Noble, and 0.50% Difco Bacto Peptone. Preferably the agar and peptone are prepared at 4× concentration in sample water, sterilized and then diluted with filter-sterilized sample water to the appropriate volume. When necessary, the media are...

example 3

[0054]Growth and isolation of novel microorganisms: Utilization of the peptide Asp-Ile-Cys-Asn-Ala-Tyr-Phe to inhibit the Gram positive, peptide-based signaling system.

[0055]In this example soil is used as the source of microorganisms, however any source that is expected to exhibit a diversity of novel (i.e., not yet cultured) microorganisms may be used.

[0056]Soil samples (approximately 500 g) are obtained from the Oh and Ah horizons of a decidous forest; the samples are maintained at 4 C until used.

[0057]Portions of the soil samples are washed 1:1 in tap water, filtered through Whatman #1 filter paper to obtain “soil water” and filter-sterilized. Agar media are prepared consisting of 1) filter-sterilized soil water, 0.5% Difco Agar Noble (Becton Dickinson, Sparks, Md.), and 0.05% Difco Bacto Peptone and 2) filter-sterilized soil water, 0.5% Difco Agar Noble, and 0.50% Difco Bacto Peptone. Preferably the agar and peptone are prepared at 4× concentration in soil water, sterilized and...

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PUM

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Abstract

A method of interfering with quorum sensing regulation of genes to promote cell growth is disclosed. The method of is aimed at accessing microbial biodiversity. The method involves obtaining an environmental sample comprising at least one novel (uncultivated in the laboratory) microorganism, contacting the environmental sample with an effective amount of an agent or combination of agents which interferes with the quorum sensing regulation of genes, growing the treated sample in a culture medium containing the quorum sensing signal disrupting agent or agents, and analyzing the colonies of microorganisms grown to demonstrate genetic novelty.

Description

BACKGROUND OF THE INVENTION[0001]1. Field of the Invention[0002]This invention relates to methods for accessing microbial diversity through disruption of microbial quorum sensing systems. In particular, this invention enables the isolation of novel microorganisms by dis-enabling quorum sensing systems that are used to maintain microbial cell density at a low level.[0003]2. Description of Related Art[0004]With recent developments in PCR technology and comparative microbial genome sequencing, it has been demonstrated in many environments that the number of microorganisms that have been cultured represents only a percentage of those present in a particular environment. It has been estimated that only approximately 1-5% of existing microorganisms have been cultured in the laboratory.[0005]The organisms which remain “uncultivated” represent a potentially large pool of genes comprising novel microbial diversity. Accessing this diversity would allow the identification for example of enzyme...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/04C12Q1/44C12Q1/37C12Q1/34G01N33/50C12N15/09A01N61/00A61K39/395C07H21/02C07K1/00C12N1/20C12N5/00C12Q1/00
CPCC12Q1/04C12Q1/44C12Q1/37C12Q1/24
Inventor KUHNER, CARLA H.MARRS, BARRYROMESSER, JAMES A.
Owner FRAUNHOFER USA
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