2001 Technology for a Sustainable Environment: NSF/EPA Partnership: Combinatorial Mutagenesis of a Bidirectional Hydrogenase in Chlamydomonas reinhardtii (TSE01-D)

Colorado School Of Mines, Golden CO

Investigators

Abstract

0124821 Ahmann The objective of this project is to perform combinatorial mutagenesis to improve the functioning of a Fe-hydrogenase for the production of hydrogen gas from algae in the presence of oxygen. The specific components of the research include: (1) the Fe-hydrogenase sequences of multiple organisms will be used as the basis for a PCR-mediated multi-parent shuffle to create recombinant hydrogenase sequences, (2) these sequences will be cloned into a Chlamydomonas vector, the vectors will be transformed into C. reinhardtii, and transformants for oxygen-resistant hydrogen production will be selected using a chemical selection procedure, and (3) a subset of the survivors will be selected using chemochromic and gas chromatographic hydrogen production assays. The goal of this research is an oxygen tolerant Fe-hydrogenase that may ultimately combined with other oxygen-inhibition-elimination mechanisms to facilitate commercial algal hydrogen production.