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

0124821Ahmann这个项目的目标是进行组合诱变，以提高在氧气存在下从藻类中生产氢气的铁氢酶的功能。这项研究的具体组成部分包括：(1)多个生物的铁氢酶序列将被用作PCR介导的多亲本改组的基础，以创建重组氢酶序列，(2)这些序列将被克隆到衣藻载体中，载体将被转化到莱茵梭菌中，并将通过化学选择程序选择耐氧产氢转化子，以及(3)将使用化学显色和气相色谱产氢分析来选择幸存者的子集。本研究的目标是一种耐氧铁氢酶，它最终可能与其他氧抑制消除机制结合起来，促进商业藻类氢气的生产。

0124821AhmannThe 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.