Anaerobic high-throughput cultivation method for isolation of thermophiles using biomass-derived substrates.
Identifieur interne : 002C06 ( Main/Exploration ); précédent : 002C05; suivant : 002C07Anaerobic high-throughput cultivation method for isolation of thermophiles using biomass-derived substrates.
Auteurs : Scott D. Hamilton-Brehm [États-Unis] ; Tatiana A. Vishnivetskaya ; Steve L. Allman ; Jonathan R. Mielenz ; James G. ElkinsSource :
- Methods in molecular biology (Clifton, N.J.) [ 1940-6029 ] ; 2012.
Descripteurs français
- KwdFr :
- Bactéries anaérobies (croissance et développement), Bactéries anaérobies (isolement et purification), Bactéries anaérobies (métabolisme), Biomasse (MeSH), Biotechnologie (méthodes), Clostridium thermocellum (croissance et développement), Clostridium thermocellum (isolement et purification), Clostridium thermocellum (métabolisme), Cytométrie en flux (méthodes), Microbiologie de l'eau (MeSH), Pyrococcus furiosus (croissance et développement), Pyrococcus furiosus (isolement et purification), Pyrococcus furiosus (métabolisme), Techniques de culture (méthodes), Température (MeSH), Tests de criblage à haut débit (méthodes), Wyoming (MeSH).
- MESH :
- croissance et développement : Bactéries anaérobies, Clostridium thermocellum, Pyrococcus furiosus.
- isolement et purification : Bactéries anaérobies, Clostridium thermocellum, Pyrococcus furiosus.
- métabolisme : Bactéries anaérobies, Clostridium thermocellum, Pyrococcus furiosus.
- méthodes : Biotechnologie, Cytométrie en flux, Techniques de culture, Tests de criblage à haut débit.
- Biomasse, Microbiologie de l'eau, Température, Wyoming.
English descriptors
- KwdEn :
- Bacteria, Anaerobic (growth & development), Bacteria, Anaerobic (isolation & purification), Bacteria, Anaerobic (metabolism), Biomass (MeSH), Biotechnology (methods), Clostridium thermocellum (growth & development), Clostridium thermocellum (isolation & purification), Clostridium thermocellum (metabolism), Culture Techniques (methods), Flow Cytometry (methods), High-Throughput Screening Assays (methods), Pyrococcus furiosus (growth & development), Pyrococcus furiosus (isolation & purification), Pyrococcus furiosus (metabolism), Temperature (MeSH), Water Microbiology (MeSH), Wyoming (MeSH).
- MESH :
- geographic : Wyoming.
- growth & development : Bacteria, Anaerobic, Clostridium thermocellum, Pyrococcus furiosus.
- isolation & purification : Bacteria, Anaerobic, Clostridium thermocellum, Pyrococcus furiosus.
- metabolism : Bacteria, Anaerobic, Clostridium thermocellum, Pyrococcus furiosus.
- methods : Biotechnology, Culture Techniques, Flow Cytometry, High-Throughput Screening Assays.
- Biomass, Temperature, Water Microbiology.
Abstract
Flow cytometry (FCM) techniques have been developed for sorting mesophilic organisms, but the difficulty increases if the target microbes are thermophilic anaerobes. We demonstrate a reliable, high-throughput method of screening thermophilic anaerobic organisms using FCM and 96-well plates for growth on biomass-relevant substrates. The method was tested using the cellulolytic thermophiles Clostridium thermocellum (T(opt) = 55 °C), Caldicellulosiruptor obsidiansis (T(opt) = 78 °C) and the fermentative hyperthermophiles, Pyrococcus furiosus (T(opt) = 100 °C) and Thermotoga maritima (T(opt) = 80 °C). Multi-well plates were incubated at various temperatures for approximately 72-120 h and then tested for growth. Positive growth resulting from single cells sorted into individual wells containing an anaerobic medium was verified by OD(600). Depending on the growth substrate, up to 80 % of the wells contained viable cultures, which could be transferred to fresh media. This method was used to isolate thermophilic microbes from Rabbit Creek, Yellowstone National Park (YNP), Wyoming. Substrates for enrichment cultures including crystalline cellulose (Avicel), xylan (from Birchwood), pretreated switchgrass and Populus were used to cultivate organisms that may be of interest to lignocellulosic biofuel production.
DOI: 10.1007/978-1-61779-956-3_15
PubMed: 22843398
Affiliations:
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Mielenz</name></author><author><name sortKey="Elkins, James G" sort="Elkins, James G" uniqKey="Elkins J" first="James G" last="Elkins">James G. Elkins</name></author></analytic><series><title level="j">Methods in molecular biology (Clifton, N.J.)</title><idno type="eISSN">1940-6029</idno><imprint><date when="2012" type="published">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Bacteria, Anaerobic (growth & development)</term><term>Bacteria, Anaerobic (isolation & purification)</term><term>Bacteria, Anaerobic (metabolism)</term><term>Biomass (MeSH)</term><term>Biotechnology (methods)</term><term>Clostridium thermocellum (growth & development)</term><term>Clostridium thermocellum (isolation & purification)</term><term>Clostridium thermocellum (metabolism)</term><term>Culture Techniques (methods)</term><term>Flow Cytometry (methods)</term><term>High-Throughput Screening Assays (methods)</term><term>Pyrococcus furiosus (growth & development)</term><term>Pyrococcus furiosus (isolation & purification)</term><term>Pyrococcus furiosus (metabolism)</term><term>Temperature (MeSH)</term><term>Water Microbiology (MeSH)</term><term>Wyoming (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Bactéries anaérobies (croissance et développement)</term><term>Bactéries anaérobies (isolement et purification)</term><term>Bactéries anaérobies (métabolisme)</term><term>Biomasse (MeSH)</term><term>Biotechnologie (méthodes)</term><term>Clostridium thermocellum (croissance et développement)</term><term>Clostridium thermocellum (isolement et purification)</term><term>Clostridium thermocellum (métabolisme)</term><term>Cytométrie en flux (méthodes)</term><term>Microbiologie de l'eau (MeSH)</term><term>Pyrococcus furiosus (croissance et développement)</term><term>Pyrococcus furiosus (isolement et purification)</term><term>Pyrococcus furiosus (métabolisme)</term><term>Techniques de culture (méthodes)</term><term>Température (MeSH)</term><term>Tests de criblage à haut débit (méthodes)</term><term>Wyoming (MeSH)</term></keywords><keywords scheme="MESH" type="geographic" xml:lang="en"><term>Wyoming</term></keywords><keywords scheme="MESH" qualifier="croissance et développement" xml:lang="fr"><term>Bactéries anaérobies</term><term>Clostridium thermocellum</term><term>Pyrococcus furiosus</term></keywords><keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Bacteria, Anaerobic</term><term>Clostridium thermocellum</term><term>Pyrococcus furiosus</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Bacteria, Anaerobic</term><term>Clostridium thermocellum</term><term>Pyrococcus furiosus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Bactéries anaérobies</term><term>Clostridium thermocellum</term><term>Pyrococcus furiosus</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Bacteria, Anaerobic</term><term>Clostridium thermocellum</term><term>Pyrococcus furiosus</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Biotechnology</term><term>Culture Techniques</term><term>Flow Cytometry</term><term>High-Throughput Screening Assays</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Bactéries anaérobies</term><term>Clostridium thermocellum</term><term>Pyrococcus furiosus</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Biotechnologie</term><term>Cytométrie en flux</term><term>Techniques de culture</term><term>Tests de criblage à haut débit</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Biomass</term><term>Temperature</term><term>Water Microbiology</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Biomasse</term><term>Microbiologie de l'eau</term><term>Température</term><term>Wyoming</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Flow cytometry (FCM) techniques have been developed for sorting mesophilic organisms, but the difficulty increases if the target microbes are thermophilic anaerobes. We demonstrate a reliable, high-throughput method of screening thermophilic anaerobic organisms using FCM and 96-well plates for growth on biomass-relevant substrates. The method was tested using the cellulolytic thermophiles Clostridium thermocellum (T(opt) = 55 °C), Caldicellulosiruptor obsidiansis (T(opt) = 78 °C) and the fermentative hyperthermophiles, Pyrococcus furiosus (T(opt) = 100 °C) and Thermotoga maritima (T(opt) = 80 °C). Multi-well plates were incubated at various temperatures for approximately 72-120 h and then tested for growth. Positive growth resulting from single cells sorted into individual wells containing an anaerobic medium was verified by OD(600). Depending on the growth substrate, up to 80 % of the wells contained viable cultures, which could be transferred to fresh media. This method was used to isolate thermophilic microbes from Rabbit Creek, Yellowstone National Park (YNP), Wyoming. Substrates for enrichment cultures including crystalline cellulose (Avicel), xylan (from Birchwood), pretreated switchgrass and Populus were used to cultivate organisms that may be of interest to lignocellulosic biofuel production.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">22843398</PMID><DateCompleted><Year>2012</Year><Month>12</Month><Day>03</Day></DateCompleted><DateRevised><Year>2012</Year><Month>07</Month><Day>30</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1940-6029</ISSN><JournalIssue CitedMedium="Internet"><Volume>908</Volume><PubDate><Year>2012</Year></PubDate></JournalIssue><Title>Methods in molecular biology (Clifton, N.J.)</Title><ISOAbbreviation>Methods Mol Biol</ISOAbbreviation></Journal><ArticleTitle>Anaerobic high-throughput cultivation method for isolation of thermophiles using biomass-derived substrates.</ArticleTitle><Pagination><MedlinePgn>153-68</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/978-1-61779-956-3_15</ELocationID><Abstract><AbstractText>Flow cytometry (FCM) techniques have been developed for sorting mesophilic organisms, but the difficulty increases if the target microbes are thermophilic anaerobes. We demonstrate a reliable, high-throughput method of screening thermophilic anaerobic organisms using FCM and 96-well plates for growth on biomass-relevant substrates. The method was tested using the cellulolytic thermophiles Clostridium thermocellum (T(opt) = 55 °C), Caldicellulosiruptor obsidiansis (T(opt) = 78 °C) and the fermentative hyperthermophiles, Pyrococcus furiosus (T(opt) = 100 °C) and Thermotoga maritima (T(opt) = 80 °C). Multi-well plates were incubated at various temperatures for approximately 72-120 h and then tested for growth. Positive growth resulting from single cells sorted into individual wells containing an anaerobic medium was verified by OD(600). Depending on the growth substrate, up to 80 % of the wells contained viable cultures, which could be transferred to fresh media. This method was used to isolate thermophilic microbes from Rabbit Creek, Yellowstone National Park (YNP), Wyoming. Substrates for enrichment cultures including crystalline cellulose (Avicel), xylan (from Birchwood), pretreated switchgrass and Populus were used to cultivate organisms that may be of interest to lignocellulosic biofuel production.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Hamilton-Brehm</LastName><ForeName>Scott D</ForeName><Initials>SD</Initials><AffiliationInfo><Affiliation>Oak Ridge National Laboratory, BioEnergy Science Center, Oak Ridge, TN, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Vishnivetskaya</LastName><ForeName>Tatiana A</ForeName><Initials>TA</Initials></Author><Author ValidYN="Y"><LastName>Allman</LastName><ForeName>Steve L</ForeName><Initials>SL</Initials></Author><Author ValidYN="Y"><LastName>Mielenz</LastName><ForeName>Jonathan R</ForeName><Initials>JR</Initials></Author><Author ValidYN="Y"><LastName>Elkins</LastName><ForeName>James G</ForeName><Initials>JG</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013486">Research Support, U.S. Gov't, Non-P.H.S.</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Methods Mol Biol</MedlineTA><NlmUniqueID>9214969</NlmUniqueID><ISSNLinking>1064-3745</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D001421" MajorTopicYN="N">Bacteria, Anaerobic</DescriptorName><QualifierName UI="Q000254" MajorTopicYN="N">growth & development</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018533" MajorTopicYN="N">Biomass</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001709" MajorTopicYN="N">Biotechnology</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D048013" MajorTopicYN="N">Clostridium thermocellum</DescriptorName><QualifierName UI="Q000254" MajorTopicYN="N">growth & development</QualifierName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D046508" MajorTopicYN="N">Culture Techniques</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005434" MajorTopicYN="N">Flow Cytometry</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D057166" MajorTopicYN="N">High-Throughput Screening Assays</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D020027" MajorTopicYN="N">Pyrococcus furiosus</DescriptorName><QualifierName UI="Q000254" MajorTopicYN="N">growth & development</QualifierName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013696" MajorTopicYN="N">Temperature</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014871" MajorTopicYN="Y">Water Microbiology</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014959" MajorTopicYN="N" Type="Geographic">Wyoming</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2012</Year><Month>7</Month><Day>31</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2012</Year><Month>7</Month><Day>31</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2012</Year><Month>12</Month><Day>10</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">22843398</ArticleId><ArticleId IdType="doi">10.1007/978-1-61779-956-3_15</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>États-Unis</li></country><region><li>Tennessee</li></region></list><tree><noCountry><name sortKey="Allman, Steve L" sort="Allman, Steve L" uniqKey="Allman S" first="Steve L" last="Allman">Steve L. Allman</name><name sortKey="Elkins, James G" sort="Elkins, James G" uniqKey="Elkins J" first="James G" last="Elkins">James G. Elkins</name><name sortKey="Mielenz, Jonathan R" sort="Mielenz, Jonathan R" uniqKey="Mielenz J" first="Jonathan R" last="Mielenz">Jonathan R. Mielenz</name><name sortKey="Vishnivetskaya, Tatiana A" sort="Vishnivetskaya, Tatiana A" uniqKey="Vishnivetskaya T" first="Tatiana A" last="Vishnivetskaya">Tatiana A. Vishnivetskaya</name></noCountry><country name="États-Unis"><region name="Tennessee"><name sortKey="Hamilton Brehm, Scott D" sort="Hamilton Brehm, Scott D" uniqKey="Hamilton Brehm S" first="Scott D" last="Hamilton-Brehm">Scott D. Hamilton-Brehm</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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