Page 7 of 8
Journal of the American Chemical Society
1
2
3
4
ize the formyl group of formyl-CoA enabling proton abstraction by Glu449 leading to the formation of CO and CoAS-. Protonated
Glu449 then re-protonates CoAS regenerating CoA.
(5) Reissmann, S.; Hochleitner, E.; Wang, H.; Paschos, A.; Lottspeich,
F.; Glass, R. S.; Böck, A. Taming of a poison: biosynthesis of the NiFe-
5
6
7
8
hydrogenase cyanide ligands. Science 2003, 299, 1067–1070.
ASSOCIATED CONTENT
(6) Forzi, L.; Hellwig, P.; Thauer, R. K.; Sawers, R. G. The CO and CN-
ligands to the active site Fe in [NiFe]-hydrogenase of Escherichia coli
have different metabolic origins. FEBS Lett 2007, 581, 3317–3321.
(7) Bürstel, I.; Siebert, E.; Winter, G.; Hummel, P.; Zebger, I.; Frie-
drich, B.; Lenz, O. A universal scaffold for synthesis of the Fe(CN)2(CO)
moiety of [NiFe] hydrogenase. J Biol Chem 2012, 287, 38845–38853.
(8) Stripp, S. T.; Soboh, B.; Lindenstrauss, U.; Braussemann, M.;
Herzberg, M.; Nies, D. H.; Sawers, R. G.; Heberle, J. HypD is the scaffold
protein for Fe-(CN)2CO cofactor assembly in [NiFe]-hydrogenase
maturation. Biochemistry 2013, 52, 3289–3296.
(9) Khorasani-Motlagh, M.; Noroozifar, M.; Kerman, K.; Zamble, D.
B. Complex formation between the Escherichia coli [NiFe]-
hydrogenase nickel maturation factors. BioMetals 2019, 32, 521–532.
(10) Bürstel, I.; Hummel, P.; Siebert, E.; Wisitruangsakul, N.; Zebger,
I.; Friedrich, B.; Lenz, O. Probing the origin of the metabolic precursor
of the CO ligand in the catalytic center of [NiFe] hydrogenase. J Biol
Chem 2011, 286, 44937–44944.
(11) Soboh, B.; Stripp, S. T.; Bielak, C.; Lindenstrauß, U.;
Braussemann, M.; Javaid, M.; Hallensleben, M.; Granich, C.; Herzberg,
M.; Heberle, J.; Sawers, R. G. The [NiFe]-hydrogenase accessory chap-
erones HypC and HybG of Escherichia coli are iron- and carbon diox-
ide-binding proteins. FEBS Lett 2013, 587, 2512–2516.
(12) Lenz, O.; Lauterbach, L.; Frielingsdorf, S.; Friedrich, B. Oxygen-
tolerant hydrogenases and their biotechnological potential. In Biohy-
drogen; Rögner, M., Ed.; Walter de Gruyter GmbH & Co. KG: Berlin,
Boston, 2015; pp 61–88.
(13) Buhrke, T.; Friedrich, B. hoxX (hypX) is a functional member of
the Alcaligenes eutrophus hyp gene cluster. Arch Microbiol 1998, 170,
460–463.
(14) Bürstel, I.; Siebert, E.; Frielingsdorf, S.; Zebger, I.; Friedrich, B.;
Lenz, O. CO synthesized from the central one-carbon pool as source
for the iron carbonyl in O2-tolerant [NiFe]-hydrogenase. Proc Natl
Acad Sci USA 2016, 113, 14722–14726.
(15) Rey, L.; Fernández, D.; Brito, B.; Hernando, Y.; Palacios, J.-M.;
Ruiz-Argüeso, T.; Imperial, J. The hydrogenase gene cluster of Rhizo-
bium leguminosarum bv. viciae contains an additional gene (hypX),
which encodes a protein with sequence similarity to the N10-
formyltetrahydrofolate-dependent enzyme family and is required for
nickel-dependent hydrogenase processing and activity. Molec Gen
Genet 1996, 252, 237–248.
(16) Muraki, N.; Ishii, K.; Uchiyama, S.; Itoh, S. G.; Okumura, H.;
Aono, S. Structural characterization of HypX responsible for CO bio-
synthesis in the maturation of NiFe-hydrogenase. Commun Biol 2019,
2, 1–12.
Supporting Information
The Supporting Information is available free of charge on the
ACS Publications website.
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Experimental details and analytical data (PDF)
AUTHOR INFORMATION
Corresponding Author
*E-mail: oliver.lenz@tu-berlin.de
ORCID
Stefan Frielingsdorf: 0000-0002-4141-7836
Phillip Pommerening: 0000-0003-2595-9448
Lars Lauterbach: 0000-0002-6601-6473
Giovanni Bistoni: 0000-0003-4849-1323
Frank Neese: 0000-0003-4691-0547
Martin Oestreich: 0000-0002-1487-9218
Oliver Lenz: 0000-0003-4550-5128
Notes
The authors declare no competing financial interests.
ACKNOWLEDGMENTS
A.C.S and O.L. gratefully acknowledge funding from the
Deutsche Forschungsgemeinschaft (DFG, German Research
Foundation) via SPP 1927 “Iron Sulfur for Life” (Project No. LE
2934/1-1). S.F., L.L., and O.L. received funding from the
Deutsche Forschungsgemeinschaft (DFG, German Research
Foundation) under Germany´s Excellence Strategy – EXC
2008/1
Forschungsgemeinschaft
–
390540038 (Gefördert durch die Deutsche
(DFG) im Rahmen der
Exzellenzstrategie des Bundes und der Länder – EXC 2008/1
– 390540038.) and the Einstein Center of Catalysis (EC2). We
are grateful to Jasmin Kurtzke and Silke Leimkühler (Universi-
tät Potsdam) for metal determination by ICP-OES. We are
indebted to Matthias Boll (Albert-Ludwigs-Universität Frei-
burg) and Bernhard Schink (Universität Konstanz) for sharing
expertise and protocols on the chemical synthesis of formyl-
CoA. We thank Georg Fuchs (Albert-Ludwigs-Universität Frei-
burg) for discussions about the phylogenetic distribution of
HypX-like proteins.
(17) Kleihues, L.; Lenz, O.; Bernhard, M.; Buhrke, T.; Friedrich, B.
The H2 sensor of Ralstonia eutropha is a member of the subclass of
regulatory [NiFe] hydrogenases. J Bacteriol 2000, 182, 2716–2724.
(18) Simon, R.; Priefer, U.; Pühler, A. A broad host range mobiliza-
tion system for in vivo genetic engineering: transposon mutagenesis in
gram negative bacteria. Nat Biotechnol 1983, 1, 784–791.
(19) Nakamura, K.; Ishida, H.; Iizumi, T. Constitutive trichloroeth-
ylene degradation led by tac promoter chromosomally integrated
upstream of phenol hydroxylase genes of Ralstonia sp. KN1 and its
nucleotide sequence analysis. J Biosci Bioeng 2000, 89, 47–54.
(20) Hartmann, S.; Frielingsdorf, S.; Ciaccafava, A.; Lorent, C.;
Fritsch, J.; Siebert, E.; Priebe, J.; Haumann, M.; Zebger, I.; Lenz, O. O2-
tolerant H2 activation by an isolated large subunit of a [NiFe] hydro-
genase. Biochemistry 2018, 57, 5339–5349.
REFERENCES
(1) Lubitz, W.; Ogata, H.; Rüdiger, O.; Reijerse, E. Hydrogenases.
Chem Rev 2014, 114, 4081–4148.
(2) Böck, A.; King, P. W.; Blokesch, M.; Posewitz, M. C. Maturation of
hydrogenases. Adv Microb Physiol 2006, 51, 1–71.
(3) Bai, L.; Fujishiro, T.; Huang, G.; Koch, J.; Takabayashi, A.; Yokono,
M.; Tanaka, A.; Xu, T.; Hu, X.; Ermler, U.; Shima, S. Towards artificial
methanogenesis: biosynthesis of the [Fe]-hydrogenase cofactor and
characterization of the semi-synthetic hydrogenase. Faraday Discuss
2017, 198, 37–58.
(4) Kuchenreuther, J. M.; Myers, W. K.; Stich, T. A.; George, S. J.;
Nejatyjahromy, Y.; Swartz, J. R.; Britt, R. D. A radical intermediate in
tyrosine scission to the CO and CN- ligands of FeFe hydrogenase.
Science 2013, 342, 472–475.
(21) Antonini, E.; Brunori, M. Hemoglobin and myoglobin in their
reactions with ligands; Amsterdam : North-Holland, 1971.
(22) Shepard, E. M.; Duffus, B. R.; George, S. J.; McGlynn, S. E.; Chal-
land, M. R.; Swanson, K. D.; Roach, P. L.; Cramer, S. P.; Peters, J. W.;
Broderick, J. B. [FeFe]-hydrogenase maturation: HydG-catalyzed
synthesis of carbon monoxide. J Am Chem Soc 2010, 132, 9247–9249.
(23) Esselborn, J.; Lambertz, C.; Adamska-Venkates, A.; Simmons,
T.; Berggren, G.; Noth, J.; Siebel, J.; Hemschemeier, A.; Artero, V.; Rei-
jerse, E.; Fontecave, M.; Lubitz, W.; Happe, T. Spontaneous activation
7
ACS Paragon Plus Environment