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Applied and Environmental Microbiology, March 2007, p. 1653-1658, Vol. 73, No. 5
0099-2240/07/$08.00+0     doi:10.1128/AEM.01827-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

A Portable Anaerobic Microbioreactor Reveals Optimum Growth Conditions for the Methanogen Methanosaeta concilii{triangledown}

Benjamin Steinhaus,1 Marcelo L. Garcia,2 Amy Q. Shen,1 and Largus T. Angenent2*

Department of Mechanical and Aerospace Engineering, Washington University in St. Louis, St. Louis, Missouri 63130,1 Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis, St. Louis, Missouri 631302

Received 2 August 2006/ Accepted 30 December 2006

Conventional studies of the optimum growth conditions for methanogens (methane-producing, obligate anaerobic archaea) are typically conducted with serum bottles or bioreactors. The use of microfluidics to culture methanogens allows direct microscopic observations of the time-integrated response of growth. Here, we developed a microbioreactor (µBR) with ~1-µl microchannels to study some optimum growth conditions for the methanogen Methanosaeta concilii. The µBR is contained in an anaerobic chamber specifically designed to place it directly onto an inverted light microscope stage while maintaining a N2-CO2 environment. The methanogen was cultured for months inside microchannels of different widths. Channel width was manipulated to create various fluid velocities, allowing the direct study of the behavior and responses of M. concilii to various shear stresses and revealing an optimum shear level of ~20 to 35 µPa. Gradients in a single microchannel were then used to find an optimum pH level of 7.6 and an optimum total NH4-N concentration of less than 1,100 mg/liter (<47 mg/liter as free NH3-N) for M. concilii under conditions of the previously determined ideal shear stress and pH and at a temperature of 35°C.

* Corresponding author. Mailing address: Washington University in St. Louis, One Brookings Drive, Campus Box 1180, St. Louis, MO 63130. Phone: (314) 935-5663. Fax: (314) 935-5464. E-mail: angenent{at}seas.wustl.edu.

{triangledown} Published ahead of print on 12 January 2007.

Applied and Environmental Microbiology, March 2007, p. 1653-1658, Vol. 73, No. 5
0099-2240/07/$08.00+0     doi:10.1128/AEM.01827-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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