[FeFe]-Hydrogenase with Chalcogenide Substitutions at the H-Cluster Maintains Full H2 Evolution Activity

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• 作者：Dr. Jens Noth ; Dr. Julian Esselborn ; Dr. Jö ; rn G&#252 ; ldenhaupt ; Annika Br&#252 ; nje ; Dr. Anne Sawyer ; Dr. Ulf-Peter Apfel ; Prof. Klaus Gerwert ; Prof. Eckhard Hofmann ; Dr. Martin Winkler and Prof. Thomas Happe
• 刊名：Angewandte Chemie International Edition
• 出版年：2016
• 出版时间：July 11, 2016
• 年：2016
• 卷：55
• 期：29
• 页码：8396-8400
• 全文大小：1660K
• ISSN：1521-3773

文摘

The [FeFe]-hydrogenase HYDA1 from Chlamydomonas reinhardtii is particularly amenable to biochemical and biophysical characterization because the H-cluster in the active site is the only inorganic cofactor present. Herein, we present the complete chemical incorporation of the H-cluster into the HYDA1-apoprotein scaffold and, furthermore, the successful replacement of sulfur in the native [4Fe_H] cluster with selenium. The crystal structure of the reconstituted pre-mature HYDA1[4Fe4Se]_H protein was determined, and a catalytically intact artificial H-cluster variant was generated upon in vitro maturation. Full hydrogen evolution activity as well as native-like composition and behavior of the redesigned enzyme were verified through kinetic assays, FTIR spectroscopy, and X-ray structure analysis. These findings reveal that even a bioinorganic active site with exceptional complexity can exhibit a surprising level of compositional plasticity.