A cell-free microtiter plate screen for improved [FeFe] hydrogenases.

ABSTRACT:

Background

[FeFe] hydrogenase enzymes catalyze the production and dissociation of H(2), a potential renewable fuel. Attempts to exploit these catalysts in engineered systems have been hindered by the biotechnologically inconvenient properties of the natural enzymes, including their extreme oxygen sensitivity. Directed evolution has been used to improve the characteristics of a range of natural catalysts, but has been largely unsuccessful for [FeFe] hydrogenases because of a lack of convenient screening platforms.

Methodology/principal findings

Here we describe an in vitro screening technology for oxygen-tolerant and highly active [FeFe] hydrogenases. Despite the complexity of the protocol, we demonstrate a level of reproducibility that allows moderately improved mutants to b

SUBMITTER: Stapleton JA 

PROVIDER: S-EPMC2866662 | biostudies-literature | 2010 May

REPOSITORIES: biostudies-literature