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[FeFe]‐Hydrogenase with Chalcogenide Substitutions at the H‐Cluster Maintains Full H 2 Evolution Activity
Author(s) -
Noth Jens,
Esselborn Julian,
Güldenhaupt Jörn,
Brünje Annika,
Sawyer Anne,
Apfel UlfPeter,
Gerwert Klaus,
Hofmann Eckhard,
Winkler Martin,
Happe Thomas
Publication year - 2016
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201511896
Subject(s) - hydrogenase , chlamydomonas reinhardtii , chemistry , bioinorganic chemistry , cluster (spacecraft) , cofactor , active site , crystallography , chalcogenide , enzyme , stereochemistry , biochemistry , organic chemistry , computer science , mutant , programming language , gene
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.