Engineering an [FeFe]-Hydrogenase: Do Accessory Clusters Influence O2 Resistance and Catalytic Bias?

被引：52

作者：

Caserta, Giorgio ^{[1
,6
]}

Papini, Cecilia ^{[1
]}

Adamska-Venkatesh, Agnieszka ^{[2
]}

Pecqueur, Ludovic ^{[1
]}

Sommer, Constanze ^{[2
]}

Reijerse, Edward ^{[2
]}

Lubitz, Wolfgang ^{[2
]}

Gauquelin, Charles ^{[3
]}

Meynial-Salles, Isabelle ^{[3
]}

Pramanik, Debajyoti ^{[4
]}

Artero, Vincent ^{[4
]}

Atta, Mohamed ^{[4
]}

del Barrio, Melisa ^{[5
]}

Faivre, Bruno ^{[1
]}

Fourmond, Vincent ^{[5
]}

Leger, Christophe ^{[5
]}

Fontecave, Marc ^{[1
]}

机构：

[1] PSL Res Univ, Univ Pierre & Marie Curie, Lab Chim Proc Biol, Coll France,CNRS,UMR 8229, 11 Pl Marcelin Berthelot, F-75005 Paris, France

[2] Max Planck Inst Chem Energiekonvers, Stiftstr 34-36, D-45470 Mulheim, Germany

[3] Univ Toulouse, CNRS, INRA, LISBP,INSA, Toulouse, France

[4] Univ Grenoble Alpes, Lab Chim & Biol Metaux, CNRS, CEA BIG, 17 Rue Martyrs, F-38000 Grenoble, France

[5] Aix Marseille Univ, CNRS, UMR 7281, Lab Bioenerget & Ingn Prot, F-13400 Marseille, France

[6] Tech Univ Berlin, Dept Chem, Max Volmer Lab, Biochem Gas Converting Biocatalysts, Str 17 Juni 135, D-10623 Berlin, Germany

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2018年 / 140卷 / 16期

关键词：

H-CLUSTER; FEFE HYDROGENASES; ACTIVE-SITE; MEGASPHAERA-ELSDENII; ESCHERICHIA-COLI; DIRECT ELECTROCHEMISTRY; IRON HYDROGENASE; ENZYMES; INACTIVATION; ACTIVATION;

D O I：

10.1021/jacs.8b01689

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

[FeFe]-hydrogenases, HydAs, are unique biocatalysts for proton reduction to H-2. However, they suffer from a number of drawbacks for biotechnological applications: size, number and diversity of metal cofactors, oxygen sensitivity. Here we show that HydA from Megasphaera elsdenii (MeHydA) displays significant resistance to O-2. Furthermore, we produced a shorter version of this enzyme (MeH-HydA), lacking the N-terminal domain harboring the accessory FeS clusters. As shown by detailed spectroscopic and biochemical characterization, MeH-HydA displays the following interesting properties. First, a functional active site can be assembled in MeH-HydA in vitro, providing the enzyme with excellent hydrogenase activity. Second, the resistance of MeHydA to O-2 is conserved in MeH-HydA. Third, MeH-HydA is more biased toward proton reduction than MeHydA, as the result of the truncation changing the rate limiting steps in catalysis. This work shows that it is possible to engineer HydA to generate an active hydrogenase that combines the resistance of the most resistant HydAs and the simplicity of algal HydAs, containing only the H-cluster.

引用

页码：5516 / 5526

页数：11

共 54 条

[1] Understanding and Tuning the Catalytic Bias of Hydrogenase [J].

Abou Hamdan, Abbas ;

Dementin, Sebastien ;

Liebgott, Pierre-Pol ;

Gutierrez-Sanz, Oscar ;

Richaud, Pierre ;

De Lacey, Antonio L. ;

Roussett, Marc ;

Bertrand, Patrick ;

Cournac, Laurent ;

Leger, Christophe .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2012, 134 (20) :8368-8371

[2] Spectroscopic Characterization of the Bridging Amine in the Active Site of [FeFe] Hydrogenase Using lsotopologues of the H-Cluster [J].

Adamska-Venkatesh, Agnieszka ;

Roy, Souvik ;

Siebel, Judith F. ;

Simmons, Trevor R. ;

Fontecave, Marc ;

Artero, Vincent ;

Reijerse, Edward ;

Lubitz, Wolfgang .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2015, 137 (40) :12744-12747

[3] New Redox States Observed in [FeFe] Hydrogenases Reveal Redox Coupling Within the H-Cluster [J].

Adamska-Venkatesh, Agnieszka ;

Krawietz, Danuta ;

Siebel, Judith ;

Weber, Katharina ;

Happe, Thomas ;

Reijerse, Edward ;

Lubitz, Wolfgang .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2014, 136 (32) :11339-11346

[4] Deletion of iscR stimulates recombinant clostridial Fe-Fe hydrogenase activity and H2-accumulation in Escherichia coli BL21(DE3) [J].

Akhtar, M. Kalim ;

Jones, Patrik R. .

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2008, 78 (05) :853-862

[5] From Enzyme Maturation to Synthetic Chemistry: The Case of Hydrogenases [J].

Artero, Vincent ;

Berggren, Gustav ;

Atta, Mohamed ;

Caserta, Giorgio ;

Roy, Souvik ;

Pecqueur, Ludovic ;

Fontecave, Marc .

ACCOUNTS OF CHEMICAL RESEARCH, 2015, 48 (08) :2380-2387

[6] Reduction Potentials of [FeFe]-Hydrogenase Accessory Iron-Sulfur Clusters Provide Insights into the Energetics of Proton Reduction Catalysis [J].

Artz, Jacob H. ;

Mulder, David W. ;

Ratzloff, Michael W. ;

Lubner, Carolyn E. ;

Zadvornyy, Oleg A. ;

LeVan, Axl X. ;

Williams, S. Garrett ;

Adams, Michael W. W. ;

Jones, Anne K. ;

King, Paul W. ;

Peters, John W. .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2017, 139 (28) :9544-9550

[7] Characterization of the gene encoding the [Fe]-hydrogenase from Megasphaera elsdenii [J].

Atta, M ;

Meyer, J .

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 2000, 1476 (02) :368-371

[8] Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants:: the Keio collection [J].

Baba, Tomoya ;

Ara, Takeshi ;

Hasegawa, Miki ;

Takai, Yuki ;

Okumura, Yoshiko ;

Baba, Miki ;

Datsenko, Kirill A. ;

Tomita, Masaru ;

Wanner, Barry L. ;

Mori, Hirotada .

MOLECULAR SYSTEMS BIOLOGY, 2006, 2 (1) :2006.0008

[9] Hydrogen-activating enzymes:: Activity does not correlate with oxygen sensitivity [J].

Baffert, Carole ;

Demuez, Marie ;

Cournac, Laurent ;

Burlat, Benedicte ;

Guigliarelli, Bruno ;

Bertrand, Patrick ;

Girbal, Laurence ;

Leger, Christophe .

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2008, 47 (11) :2052-2054

[10] CO Disrupts the Reduced H-Cluster of FeFe Hydrogenase. A Combined DFT and Protein Film Voltammetry Study [J].

Baffert, Carole ;

Bertini, Luca ;

Lautier, Thomas ;

Greco, Claudio ;

Sybirna, Kateryna ;

Ezanno, Pierre ;

Etienne, Emilien ;

Soucaille, Philippe ;

Bertrand, Patrick ;

Bottin, Herve ;

Meynial-Salles, Isabelle ;

De Gioia, Luca ;

Leger, Christophe .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2011, 133 (07) :2096-2099

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