Alliance for Sustainable Energy, LLC

Allaince for Sustainable Energy, LLC

Paul W King

Michael Seibert

Maria Lucia Ghirardi

Matthew C Posewitz

Sharon L Smolinski

A process for expression of active [FeFe]-hydrogenase in a host organism that does not contain either the structural gene(s) for [FeFe]-hydrogenases and/or homologues for the maturation genes HydE, HydF and HyG, comprising: cloning the structural hydrogenase gene(s) and/or the maturation genes HydE, HydF and HydG from an organisms that contains these genes into expression plasmids; transferring the plasmids into an organism that lacks a native [FeFe]-hydrogenase or that has a disrupted [FeFe]-hydrogenase and culturing it aerobically; and inducing anaerobiosis to provide [FeFe] hydrogenase biosynthesis and H?2#191 production.

Process and genes for expression and overexpression of active [FeFe] hydrogenases

The invention provides oxygen-resistant iron-hydrogenases ([Fe]-hydrogenases) for use in the production of H. Methods used in the design and engineering of the oxygen-resistant [Fe]-hydrogenases are disclosed, as are the methods of transforming and culturing appropriate host cells with the oxygen-resistant [Fe]-hydrogenases. Finally, the invention provides methods for utilizing the transformed, oxygen insensitive, host cells in the bulk production of Hin a light catalyzed reaction having water as the reactant.

Oxygen-resistant hydrogenases and methods for designing and making same

The invention provides oxygen- resistant iron-hydrogenases ([Fe]-hydrogenases) for use in the production of H2. Methods used in the design and engineering of the oxygen-resistant [Fe]-hydrogenases are disclosed, as are the methods of transforming and culturing appropriate host cells with the oxygen-resistant [Fe]-hydrogenases. Finally, the invention provides methods for utilizing the transformed, oxygen insensitive, host cells in the bulk production of Hin a light catalyzed reaction having water as the reactant.

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Golden, CO, United States of America

Paul W King