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Dissection and engineering of the Escherichia coli formate hydrogenlyase complex
Author(s) -
McDowall Jennifer S.,
Hjersing M. Charlotte,
Palmer Tracy,
Sargent Frank
Publication year - 2015
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2015.08.043
Subject(s) - escherichia coli , formate , mutagenesis , chemistry , protein subunit , protein engineering , hydrogenase , strain (injury) , computational biology , biochemistry , enzyme , combinatorial chemistry , gene , biology , mutant , anatomy , catalysis
The Escherichia coli formate hydrogenlyase (FHL) complex is produced under fermentative conditions and couples formate oxidation to hydrogen production. In this work, the architecture of FHL has been probed by analysing affinity‐tagged complexes from various genetic backgrounds. In a successful attempt to stabilize the complex, a strain encoding a fusion between FdhF and HycB has been engineered and characterised. Finally, site‐directed mutagenesis of the hycG gene was performed, which is predicted to encode a hydrogenase subunit important for regulating sensitivity to oxygen. This work helps to define the core components of FHL and provides solutions to improving the stability of the enzyme.