CA2247351A1 - Compartmentalization method for screening microorganisms - Google Patents

Compartmentalization method for screening microorganisms

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Publication number
CA2247351A1
CA2247351A1 CA002247351A CA2247351A CA2247351A1 CA 2247351 A1 CA2247351 A1 CA 2247351A1 CA 002247351 A CA002247351 A CA 002247351A CA 2247351 A CA2247351 A CA 2247351A CA 2247351 A1 CA2247351 A1 CA 2247351A1
Authority
CA
Canada
Prior art keywords
microorganisms
compartments
growth
selectable characteristic
growth chamber
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CA002247351A
Other languages
French (fr)
Other versions
CA2247351C (en
Inventor
Volker Schellenberger
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Danisco US Inc
Original Assignee
Genencor International, Inc.
Volker Schellenberger
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Genencor International, Inc., Volker Schellenberger filed Critical Genencor International, Inc.
Publication of CA2247351A1 publication Critical patent/CA2247351A1/en
Application granted granted Critical
Publication of CA2247351C publication Critical patent/CA2247351C/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/06—Quantitative determination
    • C12Q1/08—Quantitative determination using multifield media
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material
    • C12Q1/20—Testing for antimicrobial activity of a material using multifield media

Abstract

A method is provided for screening microorganisms having a selectable characteristic comprising: (a) providing a growth chamber capable of forming a plurality of discrete compartments, said compartments comprising a barrier which inhibits the diffusion of metabolites or nutrients between said compartments within said growth chamber; (b) preparing a growth medium which, by its lack of or addition of one or more metabolites or nutrients, preferentially facilitates growth of microorganisms exhibiting said selectable characteristic from microorganisms which do not exhibit said selectable characteristic; (c) inoculating said growth medium with a plurality of microorganisms, a portion of which comprises said selectable characteristic; (d) placing said inoculated growth medium into said growth chamber so as to randomly disperse said microorganisms among discrete compartments in said growth chamber, and (e) incubating said growth chamber for a suitable time and under suitable conditions to obtain a detectable disparity of growth between microorganisms having said selectable characteristic and microorganisms lacking said selectable characteristic.

Claims (20)

1. A method of screening microorganisms having a selectable characteristic comprising:
(a) providing a growth chamber capable of forming a plurality of discrete compartments, said compartments comprising a barrier which inhibits the diffusion of metabolites or nutrients between said compartments within said growth chamber;
(b) preparing a growth medium which, by its lack of or addition of one or more metabolites or nutrients, preferentially facilitates growth of microorganisms exhibiting said selectable characteristic from microorganisms which do not exhibit said selectable characteristic;
(c) inoculating said growth medium with a plurality of microorganisms a portion of which comprise said selectable characteristic;
(d) placing said inoculated growth medium into said growth chamber so as to randomly disperse said microorganisms among discrete compartments in said growthchamber;
(e) incubating said growth chamber for a suitable time and under suitable conditions to obtain a detectable disparity of growth between microorganisms having said selectable characteristic and microorganisms lacking said selectable characteristic; and (f) selecting microorganisms identified as having said selectable characteristic.
2. The method according to claim 1, wherein upon random dispersion of said pool at least 25% of said compartments contain only one microorganism.
3. The method according to claim 2, wherein 50% of said compartments contain only one microorganism.
4. The method according to claim 3, wherein 75% of said compartments contain only one microorganism.
5. The method according to claim 4, wherein substantially all of said compartments contain only one microorganism.
6. The method of claim 1, wherein said compartments comprise a volume of between about 0.1 and about one milliliter.
7. The method of claim 1, wherein said compartments comprise a volume of between about 0.001 and about 0.1 milliliter.
8. The method of claim 1, wherein said compartments comprise a volume of between about 0.0001 and about 0.01 milliliter.
9. The method of claim 1, wherein said selectable characteristic comprises a mutant enzyme exhibiting altered performance.
10. The method of claim 9, wherein said mutant enzyme exhibiting altered performance comprises altered substrate specificity, altered specific activity, altered temperature/activity profile, altered pH/activity profile, altered salt or ion requirements, altered activity or stability in the presence of surfactants, solvents, or other solutes.
11. The method of claim 9, wherein said enzyme comprises a hydrolase, an oxidoreductase, a transferase, a lyase or a ligase.
12. The method of claim 9, wherein said enzyme comprises a protease, lipase, amylase, .beta.-galactosidase, cellulase, lactase, polygalacturonase, .beta.-glucoamylase, esterase, hemicellulase, peroxidase, oxidase, laccase, glucose oxidase, ligninase NADH reductase or 2,5DKG reductase.
13. The method of claim 1, wherein said microorganisms comprise bacteria, filamentous fungi or yeast.
14. The method of claim 1, wherein said microorgansims are subjected to mutagenesis prior to inoculating said growth medium with said microorganisms.
15. The method of claim 1, wherein said growth means comprises hollow fibers, glass beads, dispersion of the inoculated media in a non-miscible fluid, or encapsulation of the media in a solid matrix.
16. A method of isolating microorganisms having a selectable characteristic comprising:
17 (a) preparing a population of microorganisms for which said selectable characteristic is desired and subjecting said population to one or more mutational events to produce mutant microorganisms;
(b) providing a growth chamber capable of forming a plurality of discrete compartments, said compartments comprising a barrier which inhibits the diffusion of metabolites or nutrients or proteins between said compartments within said growth chamber;
(c) preparing a growth medium which by its lack of or addition of one or more metabolites or nutrients, preferentially facilitates growth of said mutant microorganisms exhibiting said selectable characteristic from mutant or non-mutant microorganisms which do not exhibit said selectable characteristic;
(d) inoculating said growth medium with said mutant microorganisms;
(e) placing said inoculated growth medium into said growth chamber so as to randomly disperse said inoculated microorganisms among discrete compartments in said growth chamber;
(f) incubating said growth chamber for a suitable time and under suitable conditions to obtain a detectable disparity of growth between mutant microorganisms having said selectable characteristic and microorganisms lacking said selectable characteristic; and (g) selecting said mutant microorganisms identified as having said selectable characteristic.

17. The method according to claim 16, wherein said step (b) occurs prior to step (c).
18. The method according to claim 16, wherein said step (b) occurs simultaneously with or subsequent to step (c).
19. The method according to claim 16, wherein 50% of said compartments contain only one microorganism.
20. The method according to claim 19, wherein substantially all of said compartments contain only one microorganism.
CA2247351A 1996-03-29 1997-02-17 Compartmentalization method for screening microorganisms Expired - Lifetime CA2247351C (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US08/625,488 1996-03-29
US08/625,488 US6001586A (en) 1996-03-29 1996-03-29 Compartmentalization method for screening microorganisms
PCT/US1997/002527 WO1997037036A1 (en) 1996-03-29 1997-02-17 Compartmentalization method for screening microorganisms

Publications (2)

Publication Number Publication Date
CA2247351A1 true CA2247351A1 (en) 1997-10-09
CA2247351C CA2247351C (en) 2010-09-21

Family

ID=24506321

Family Applications (1)

Application Number Title Priority Date Filing Date
CA2247351A Expired - Lifetime CA2247351C (en) 1996-03-29 1997-02-17 Compartmentalization method for screening microorganisms

Country Status (8)

Country Link
US (1) US6001586A (en)
EP (1) EP0892853B1 (en)
AT (1) ATE278802T1 (en)
AU (1) AU731935B2 (en)
CA (1) CA2247351C (en)
DE (1) DE69731080T2 (en)
DK (1) DK0892853T3 (en)
WO (1) WO1997037036A1 (en)

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US6936289B2 (en) 1995-06-07 2005-08-30 Danisco A/S Method of improving the properties of a flour dough, a flour dough improving composition and improved food products
US6696286B1 (en) 1997-04-09 2004-02-24 3M Innovative Properties Company Method and devices for detecting and enumerating microorganisms
US6391578B2 (en) 1997-04-09 2002-05-21 3M Innovative Properties Company Method and devices for partitioning biological sample liquids into microvolumes
DE69835112T2 (en) 1997-04-09 2007-01-04 Danisco A/S Use of lipase to improve doughs and baked goods
JP4217378B2 (en) * 1997-08-01 2009-01-28 スリーエム カンパニー Methods and apparatus for detecting and counting microorganisms
US6893877B2 (en) 1998-01-12 2005-05-17 Massachusetts Institute Of Technology Methods for screening substances in a microwell array
DE69930726T2 (en) * 1998-01-12 2007-01-25 Massachusetts Institute Of Technology, Cambridge METHOD AND DEVICE FOR MICROTEST PROCESSING
ATE231186T1 (en) 1998-07-21 2003-02-15 Danisco GROCERIES
US6723504B1 (en) 1998-10-28 2004-04-20 Novozymes A/S Method for generating a gene library
US6174699B1 (en) 1999-03-09 2001-01-16 3M Innovative Properties Company Disc assay device with inoculation pad and methods of use
EP1165235B1 (en) 1999-03-19 2011-09-28 Life Technologies Corporation Method of screening mutated cells
GB9929808D0 (en) * 1999-12-16 2000-02-09 Carr Anthony H Microbial growth variations
US20020151040A1 (en) 2000-02-18 2002-10-17 Matthew O' Keefe Apparatus and methods for parallel processing of microvolume liquid reactions
US6248541B1 (en) * 2000-04-21 2001-06-19 Genencor International, Inc. Screening under nutrient limited conditions
BR0209154A (en) 2001-05-18 2004-07-20 Danisco Process of preparing a dough with an enzyme
US7011957B2 (en) 2001-09-26 2006-03-14 Northeastern University Isolation and cultivation of microorganisms from natural environments and drug discovery based thereon
US20030059866A1 (en) * 2001-09-26 2003-03-27 Kim Lewis Isolation and cultivation of microorganisms from natural environments and drug discovery based thereon
US20040137547A1 (en) * 2001-12-28 2004-07-15 Medtronic Minimed, Inc. Method for formulating a glucose oxidase enzyme with a desired property or properties and a glucose oxidase enzyme with the desired property
CN1649996A (en) * 2002-01-04 2005-08-03 宝洁公司 Cellulose-active microorganisms
US8277753B2 (en) 2002-08-23 2012-10-02 Life Technologies Corporation Microfluidic transfer pin
EP1608952B1 (en) 2002-12-20 2016-08-10 Life Technologies Corporation Assay apparatus and method using microfluidic arrays
DE602004030000D1 (en) 2003-01-17 2010-12-23 Danisco PROCESS FOR IN-SITU-PRODUCTION OF AN EMULSIFIER IN A FOODSTUFF
US20050196766A1 (en) 2003-12-24 2005-09-08 Soe Jorn B. Proteins
US7955814B2 (en) 2003-01-17 2011-06-07 Danisco A/S Method
US7417726B2 (en) 2003-09-19 2008-08-26 Applied Biosystems Inc. Normalization of data using controls
US20050221357A1 (en) 2003-09-19 2005-10-06 Mark Shannon Normalization of gene expression data
US7906307B2 (en) 2003-12-24 2011-03-15 Danisco A/S Variant lipid acyltransferases and methods of making
US7718408B2 (en) 2003-12-24 2010-05-18 Danisco A/S Method
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BRPI0720801A2 (en) 2007-01-25 2014-09-16 Dupont Nutrition Biosci Aps Production of an acyltransferase lipid from transformed cells of Bacillus licheniformis.
FR2927090A1 (en) * 2008-01-31 2009-08-07 Julien Sylvestre MASS SELECTION OF CELLS EXTRATING EXTRACELLULAR ENZYMES IN A COMPARTMENTALLY SELECTIVE MEDIUM
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WO2021168182A1 (en) * 2020-02-21 2021-08-26 Eastern Kentucky University Method for detection of escherichia coli and antibiotic resistant bacteria in water
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Also Published As

Publication number Publication date
DE69731080D1 (en) 2004-11-11
ATE278802T1 (en) 2004-10-15
AU731935B2 (en) 2001-04-05
WO1997037036A1 (en) 1997-10-09
DK0892853T3 (en) 2005-01-31
DE69731080T2 (en) 2006-02-23
CA2247351C (en) 2010-09-21
EP0892853B1 (en) 2004-10-06
US6001586A (en) 1999-12-14
EP0892853A1 (en) 1999-01-27
AU2052097A (en) 1997-10-22

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