Molecular tools for investigating ANME community structure and function

Book · Fri May 20 00:00:00 EDT 2011

OSTI ID:1023729

Hallam, Steven J; Page, Antoine P; Constan, Lea; Song, Young C; Norbeck, Angela D; Brewer, Heather M; Pasa-Tolic, Ljiljana

Methane production and consumption in anaerobic marine sediments 1 is catalyzed by a series of reversible tetramethanopterin (H4MPT)-linked C1 transfer reactions. Although many of these reactions are conserved between one-carbon compound utilizing microorganisms, two remain diagnostic for archaeal methane metabolism. These include reactions catalyzed by N5-methyltetrahydromethanopterin: coenzyme M methyltransferase and methyl coenzyme M reductase. The latter enzyme is central to C-H bond formation and cleavage underlying methanogenic and reverse methanogenic phenotypes. Here we describe a set of novel tools for the detection and functional analysis of H4MPT-linked C1 transfer reactions mediated by uncultivated anaerobic methane oxidizing archaea (ANME). These tools include polymerase chain reaction primers targeting ANME methyl coenzyme M reductase subunit A subgroups and protein extraction methods from marine sediments compatible with high-resolution mass spectrometry for profiling population structure and functional dynamics. [910, 1,043]

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 1023729
• Report Number(s): PNNL-SA-75404; 40089; 25674; KP1704020; TRN: US201120%%1065
• Resource Relation: Related Information: Methods in Enzymology, Part A, 494:75-90
• Country of Publication: United States
• Language: English

Related Subjects

03 NATURAL GAS
ANAEROBIC CONDITIONS
CATALYSIS
CLEAVAGE
COENZYMES
DETECTION
DYNAMICS
ENZYMES
EXTRACTION
FUNCTIONAL ANALYSIS
MASS SPECTROSCOPY
METABOLISM
METHANE
MICROORGANISMS
OXIDOREDUCTASES
POLYMERASE CHAIN REACTION
PRODUCTION
PROTEINS
SEDIMENTS
TRANSFER REACTIONS
Environmental Molecular Sciences Laboratory