Continuous culture of Methanococcus maripaludis under defined nutrient conditions

FEMS Microbiology Letters
Andrew K HaydockJohn A Leigh

Abstract

To study global regulation in the methanogenic archaeon Methanococcus maripaludis, we devised a system for steady-state growth in chemostats. New Brunswick Bioflo 110 bioreactors were equipped with controlled delivery of hydrogen, nitrogen, carbon dioxide, hydrogen sulfide, and anaerobic medium. We determined conditions and media compositions for growth with three different limiting nutrients, hydrogen, phosphate, and leucine. To investigate leucine limitation we constructed and characterized a mutant in the leuA gene for 2-isopropylmalate synthase, demonstrating for the first time the function of this gene in the Archaea. Steady state specific growth rates in these studies ranged from 0.042 to 0.24 h(-1). Plots of culture density vs. growth rate for each condition showed the behavior predicted by growth modeling. The results show that growth behavior is normal and reproducible and validate the use of the chemostat system for metabolic and global regulation studies in M. maripaludis.

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Citations

Jun 29, 2012·The ISME Journal·Christopher B WalkerDavid A Stahl
Jan 5, 2005·Nature Reviews. Genetics·Thorsten Allers, Moshe Mevarech
Jun 11, 2010·Proceedings of the National Academy of Sciences of the United States of America·Kyle C CostaJohn A Leigh
Jul 14, 2011·Genome Research·Sung Ho YoonNitin S Baliga
Jan 22, 2008·Journal of Bacteriology·Erik L HendricksonJohn A Leigh
May 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Erik L HendricksonJohn A Leigh
Jan 20, 2018·Archaea : an International Microbiological Journal·Feng LongWilliam B Whitman

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