title: A functional approach to uncover the low-temperature adaptation strategies of the archaeon Methanosarcina barkeri.
abstract: Low-temperature anaerobic digestion (LTAD) technology is underpinned by a diverse microbial community. The methanogenic archaea represent a key functional group in these consortia, undertaking CO2 reduction as well as acetate and methylated C1 metabolism with subsequent biogas (40 to 60% CH4 and 30 to 50% CO2) formation. However, the cold adaptation strategies, which allow methanogens to function efficiently in LTAD, remain unclear. Here, a pure-culture proteomic approach was employed to study the functional characteristics of Methanosarcina barkeri (optimum growth temperature, 37°C), which has been detected in LTAD bioreactors. Two experimental approaches were undertaken. The first approach aimed to characterize a low-temperature shock response (LTSR) of M. barkeri DSMZ 800(T) grown at 37°C with a temperature drop to 15°C, while the second experimental approach aimed to examine the low-temperature adaptation strategies (LTAS) of the same strain when it was grown at 15°C. The latter experiment employed cell viability and growth measurements (optical density at 600 nm [OD600]), which directly compared M. barkeri cells grown at 15°C with those grown at 37°C. During the LTSR experiment, a total of 127 proteins were detected in 37°C and 15°C samples, with 20 proteins differentially expressed with respect to temperature, while in the LTAS experiment 39% of proteins identified were differentially expressed between phases of growth. Functional categories included methanogenesis, cellular information processing, and chaperones. By applying a polyphasic approach (proteomics and growth studies), insights into the low-temperature adaptation capacity of this mesophilically characterized methanogen were obtained which suggest that the metabolically diverse Methanosarcinaceae could be functionally relevant for LTAD systems.
authors: Microbial Ecology Laboratory, Microbiology, School of Natural Sciences, and Ryan Institute, National University of Ireland, Galway, Ireland. e.gunnigle1@nuigalway.ie
authors: Eoin
authors: E
authors: Gunnigle
authors: Paul
authors: P
authors: McCay
authors: Matthew
authors: M
authors: Fuszard
authors: Catherine H
authors: CH
authors: Botting
authors: Florence
authors: F
authors: Abram
authors: Vincent
authors: V
authors: O'Flaherty
mesh_terms: D019342:Acetic Acid / Q000378:metabolism; D000222:Adaptation, Physiological; D001426:Bacterial Proteins / Q000378:metabolism*; D019149:Bioreactors / Q000382:microbiology; D002245:Carbon Dioxide / Q000378:metabolism; D002853:Chromatography, Liquid; D003080:Cold Temperature; D058639:Cold-Shock Response; D015180:Electrophoresis, Gel, Two-Dimensional; D006859:Hydrogen / Q000378:metabolism; D000432:Methanol / Q000378:metabolism; D017021:Methanosarcina barkeri / Q000254:growth & development / Q000502:physiology*; D020543:Proteome / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D001426:Bacterial Proteins; D020543:Proteome; D002245:Carbon Dioxide; D006859:Hydrogen; D019342:Acetic Acid; D000432:Methanol
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