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Applied and Environmental Microbiology, April 2009, p. 1867-1875, Vol. 75, No. 7
0099-2240/09/$08.00+0     doi:10.1128/AEM.02009-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Engineering a Synthetic Dual-Organism System for Hydrogen Production{triangledown}

Zeev Waks and Pamela A. Silver*

Department of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Boston, Massachusetts 02115

Received 29 August 2008/ Accepted 1 February 2009

Molecular hydrogen produced biologically from renewable biomass is an attractive replacement for fossil fuels. One potential route for biological hydrogen production is the conversion of biomass into formate, which can subsequently be processed into hydrogen by Escherichia coli. Formate is also a widely used commodity chemical, making its bioproduction even more attractive. Here we demonstrate the implementation of a formate-overproducing pathway in Saccharomyces cerevisiae, a well-established industrial organism. By expressing the anaerobic enzyme pyruvate formate lyase from E. coli, we engineered a strain of yeast that overproduced formate relative to undetectable levels in the wild type. The addition of a downstream enzyme, AdhE of E. coli, resulted in an additional 4.5-fold formate production increase as well as an increase in growth rate and biomass yield. Overall, an 18-fold formate increase was achieved in a strain background whose formate degradation pathway had been deleted. Finally, as a proof of concept, we were able to produce hydrogen from this formate-containing medium by using E. coli as a catalyst in a two-step process. With further optimizations, it may be feasible to use S. cerevisiae on a larger scale as the foundation for yeast-based biohydrogen.

* Corresponding author. Mailing address: Department of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115. Phone: (617) 432-6401. Fax: (617) 432-5012. E-mail: pamela_silver{at}hms.harvard.edu

{triangledown} Published ahead of print on 6 February 2009.

Applied and Environmental Microbiology, April 2009, p. 1867-1875, Vol. 75, No. 7
0099-2240/09/$08.00+0     doi:10.1128/AEM.02009-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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