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

2001 可持续环境技术：NSF/EPA 合作伙伴关系：莱茵衣藻双向氢化酶的组合诱变 (TSE01-D)

• 批准号: 0124821
• 负责人: Dianne Ahmann
• 金额: $ 9.96万
• 依托单位: Colorado School of Mines
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2004-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0124821&HistoricalAwards=false
• 关键词: 2001; Technology; Sustainable; Environment; NSF

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.

该项目的目的是进行组合诱变以改善Fe-氢化酶在氧气存在下从藻类产生氢气的功能。 本研究的具体内容包括：（1）以多个生物体的铁氢化酶序列为基础，通过PCR介导的多亲本改组，构建重组氢化酶序列;（2）将这些序列克隆到衣原体载体中，转化到C. reinhardtii的转化体，并使用化学选择方法选择耐氧产氢的转化体，和（3）使用化学显色和气相色谱产氢测定法选择存活者的子集。 本研究的目标是一种耐氧铁氢化酶，最终可能与其他氧抑制-消除机制相结合，以促进商业藻类产氢。