OLD YELLOW ENZYME ENGINEERING

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们正在将最先进的蛋白质工程技术应用于烯烃还原酶，使其成为制备有机合成的首选催化剂。我们选择迭代饱和诱变（ISM）用于大多数蛋白质改良研究，因为这种方法需要相对较小的文库（ca. 100）和相应的简单技术。我们正在使用X射线晶体学来评估蛋白质工程研究的结果，并指导未来的努力。两个相关的结构已经解决，其他几个结构也接近完成。 我们的首选烯烃还原酶是树干毕赤酵母OYE 2.6;如果这证明不合适，我们也准备使用巴斯德酵母（以前的卡尔斯伯酵母）OYE 1。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We are applying sstate-of-the-art protein engineering techniques to alkene reductases so that they become first-choice catalysts for preparative organic synthesis.We have chosen iterative saturation mutagenesis (ISM) for most of the protein improvement studies since this methodology requires relatively small libraries (ca. 100 for a given amino acid) and correspondingly simple technology. We are using X-ray crystallography to assess the results of protein engineering studies and guide future efforts. Two relevant structures have been solved with several others nearing completion. Our first choice alkene reductase is Pichia stipitis OYE 2.6; should this prove unsuitable, we are also prepared to use Saccharomyces pastorianus (formerly Saccharomyces carlsbergensis) OYE1.