Alliance for Sustainable Energy, LLC

Midwest Research Institute

Allaince for Sustainable Energy, LLC

Maria Lucia Ghirardi

Michael Seibert

Paul W King

Anastasios Melis

John Rudiger Benemann

Matthew C Posewitz

Marc Forestier

Sharon L Smolinski

Sergey Kosourov

Liping Zhang

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

A method of using sequential chemostat culture vessels to provide continuous Hproduction, in which photosynthetic Oevolution and Hphotoproduction are separated physically into two separate bioreactors, comprising: a) growing a microorganism culture able to continuously generate Hby photosynthetically producing cells at about the early-to-late log state in a first photobioreactor operating as a sulfur chemostat under aerobic and/or conditions; b) continuously feeding cells from the first photobioreactor to a second photobioreactor operating under anaerobic conditions and sulfur deprivation conditions resulting from constant uptake of sulfate in the first bioreactor and a low rate of culture flow between the first and second bioreactors, for induction of hydrogenase and Hphotoproduction to allow for continuous cultivation of the microorganism's cells in the first photobioreactor and constant Hproduction in the second photobioreactor, and c) Hgas from the second photobioreactor.

Multi-stage microbial system for continuous hydrogen production

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.

A reversible physiological process provides for the temporal separation of oxygen evolution and hydrogen production in a microorganism, which includes the steps of growing a culture of the microorganism in medium under illuminated conditions to accumulate an endogenous substrate, depleting from the medium a nutrient selected from the group consisting of sulfur, iron, and/or manganese, sealing the culture from atmospheric oxygen, incubating the culture in light whereby a rate of light-induced oxygen production is equal to or less than a rate of respiration, and collecting an evolved gas. The process is particularly useful to accomplish a sustained photobiological hydrogen gas production in cultures of microorganisms, such as

Hydrogen production using hydrogenase-containing oxygenic photosynthetic organisms

A process for selection of oxygen-tolerant, H.sub.2 -producing algal mutant cells comprising:

Growing community of inventors

Lakewood, CO, United States of America

Maria Lucia Ghirardi