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

[1248 . 21]这个项目的目标是进行组合诱变，以改善铁氢化酶的功能，使藻类在氧气存在的情况下产生氢气。研究的具体组成部分包括：(1)将多个生物的铁氢化酶序列作为pcr介导的多亲本洗选的基础，以创建重组氢化酶序列；(2)将这些序列克隆到衣藻载体中，将载体转化为莱因哈特氏菌，并通过化学选择程序选择抗氧产氢的转化体。(3)将使用化学变色和气相色谱产氢试验选择幸存者的一部分。本研究的目标是一种耐氧铁氢化酶，它可能最终与其他氧抑制-消除机制结合，以促进商业藻类制氢。

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.