Heterologous Expression and Engineering of the Nitrogenase Cofactor Biosynthesis Scaffold NifEN
Abstract
Abstract NifEN plays a crucial role in the biosynthesis of nitrogenase, catalyzing the final step of cofactor maturation prior to delivering the cofactor to NifDK, the catalytic component of nitrogenase. The difficulty in expressing NifEN, a complex, heteromultimeric metalloprotein sharing structural/functional homology with NifDK, is a major challenge in the heterologous expression of nitrogenase. Herein, we report the expression and engineering of Azotobacter vinelandii NifEN in Escherichia coli . Biochemical and spectroscopic analyses demonstrate the integrity of the heterologously expressed NifEN in composition and functionality and, additionally, the ability of an engineered NifEN variant to mimic NifDK in retaining the matured cofactor at an analogous cofactor‐binding site. This is an important step toward piecing together a viable pathway for the heterologous expression of nitrogenase and identifying variants for the mechanistic investigation of this enzyme.
- Authors:
-
[1];
- Univ. of California, Irvine, CA (United States). Dept. of Molecular Biology & Biochemistry. Dept. Chemistry
- Univ. of California, Irvine, CA (United States). Dept. of Molecular Biology & Biochemistry
- Publication Date:
- Research Org.:
- Univ. of California, Irvine, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1803013
- Alternate Identifier(s):
- OSTI ID: 1602596
- Grant/Contract Number:
- SC0016510
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Angewandte Chemie (International Edition)
- Additional Journal Information:
- Journal Name: Angewandte Chemie (International Edition); Journal Volume: 59; Journal Issue: 17; Journal ID: ISSN 1433-7851
- Publisher:
- Wiley
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; protein engineering; metalloproteins; metalloenzymes; nitrogenase; bioinorganic chemistry
Citation Formats
Solomon, Joseph B., Lee, Chi Chung, Jasniewski, Andrew J., Rasekh, Mahtab F., Ribbe, Markus W., and Hu, Yilin. Heterologous Expression and Engineering of the Nitrogenase Cofactor Biosynthesis Scaffold NifEN. United States: N. p., 2020.
Web. doi:10.1002/anie.201916598.
Solomon, Joseph B., Lee, Chi Chung, Jasniewski, Andrew J., Rasekh, Mahtab F., Ribbe, Markus W., & Hu, Yilin. Heterologous Expression and Engineering of the Nitrogenase Cofactor Biosynthesis Scaffold NifEN. United States. https://doi.org/10.1002/anie.201916598
Solomon, Joseph B., Lee, Chi Chung, Jasniewski, Andrew J., Rasekh, Mahtab F., Ribbe, Markus W., and Hu, Yilin. Wed .
"Heterologous Expression and Engineering of the Nitrogenase Cofactor Biosynthesis Scaffold NifEN". United States. https://doi.org/10.1002/anie.201916598. https://www.osti.gov/servlets/purl/1803013.
@article{osti_1803013,
title = {Heterologous Expression and Engineering of the Nitrogenase Cofactor Biosynthesis Scaffold NifEN},
author = {Solomon, Joseph B. and Lee, Chi Chung and Jasniewski, Andrew J. and Rasekh, Mahtab F. and Ribbe, Markus W. and Hu, Yilin},
abstractNote = {Abstract NifEN plays a crucial role in the biosynthesis of nitrogenase, catalyzing the final step of cofactor maturation prior to delivering the cofactor to NifDK, the catalytic component of nitrogenase. The difficulty in expressing NifEN, a complex, heteromultimeric metalloprotein sharing structural/functional homology with NifDK, is a major challenge in the heterologous expression of nitrogenase. Herein, we report the expression and engineering of Azotobacter vinelandii NifEN in Escherichia coli . Biochemical and spectroscopic analyses demonstrate the integrity of the heterologously expressed NifEN in composition and functionality and, additionally, the ability of an engineered NifEN variant to mimic NifDK in retaining the matured cofactor at an analogous cofactor‐binding site. This is an important step toward piecing together a viable pathway for the heterologous expression of nitrogenase and identifying variants for the mechanistic investigation of this enzyme.},
doi = {10.1002/anie.201916598},
journal = {Angewandte Chemie (International Edition)},
number = 17,
volume = 59,
place = {United States},
year = {Wed Feb 05 00:00:00 EST 2020},
month = {Wed Feb 05 00:00:00 EST 2020}
}
Free Publicly Available Full Text
Publisher's Version of Record
Other availability
Search WorldCat to find libraries that may hold this journal
Cited by: 6 works
Citation information provided by
Web of Science
Web of Science
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.
Works referenced in this record:
Identification of a nitrogenase FeMo cofactor precursor on NifEN complex
journal, February 2005
- Hu, Y.; Fay, A. W.; Ribbe, M. W.
- Proceedings of the National Academy of Sciences, Vol. 102, Issue 9
Structural insights into a protein-bound iron-molybdenum cofactor precursor
journal, January 2006
- Corbett, M. C.; Hu, Y.; Fay, A. W.
- Proceedings of the National Academy of Sciences, Vol. 103, Issue 5
Fischer-Tropsch-Chemie bei Raumtemperatur?
journal, July 2011
- Gerlach, Deidra L.; Lehnert, Nicolai
- Angewandte Chemie, Vol. 123, Issue 35
Nitrogenase MoFe-Protein at 1.16 A Resolution: A Central Ligand in the FeMo-Cofactor
journal, September 2002
- Einsle, O.
- Science, Vol. 297, Issue 5587
Optimization of FeMoco Maturation on NifEN
journal, July 2009
- Yoshizawa, Janice M.; Blank, Michael A.; Fay, Aaron W.
- Journal of the American Chemical Society, Vol. 131, Issue 26
Tuning Electron Flux through Nitrogenase with Methanogen Iron Protein Homologues
journal, October 2017
- Hiller, Caleb J.; Stiebritz, Martin T.; Lee, Chi Chung
- Chemistry - A European Journal, Vol. 23, Issue 64
The Azotobacter vinelandii NifEN Complex Contains Two Identical [4Fe-4S] Clusters †
journal, July 1998
- Goodwin, Paul J.; Agar, Jeffrey N.; Roll, Jon T.
- Biochemistry, Vol. 37, Issue 29
A fuel-producing microbe
journal, January 2017
- Dobbek, Holger
- Nature Chemical Biology, Vol. 13, Issue 2
Mechanism of Nitrogen Fixation by Nitrogenase: The Next Stage
journal, January 2014
- Hoffman, Brian M.; Lukoyanov, Dmitriy; Yang, Zhi-Yong
- Chemical Reviews, Vol. 114, Issue 8
Elucidation of the biosynthesis of the methane catalyst coenzyme F430
journal, February 2017
- Moore, Simon J.; Sowa, Sven T.; Schuchardt, Christopher
- Nature, Vol. 543, Issue 7643
Spectroscopic Characterization of an Eight-Iron Nitrogenase Cofactor Precursor that Lacks the “9 th Sulfur”
journal, September 2019
- Jasniewski, Andrew J.; Wilcoxen, Jarett; Tanifuji, Kazuki
- Angewandte Chemie International Edition, Vol. 58, Issue 41
Refactoring the nitrogen fixation gene cluster from Klebsiella oxytoca
journal, April 2012
- Temme, K.; Zhao, D.; Voigt, C. A.
- Proceedings of the National Academy of Sciences, Vol. 109, Issue 18
Structure of a Cofactor-Deficient Nitrogenase MoFe Protein
journal, April 2002
- Schmid, B.
- Science, Vol. 296, Issue 5566
Structure of Precursor-Bound NifEN: A Nitrogenase FeMo Cofactor Maturase/Insertase
journal, January 2011
- Kaiser, J. T.; Hu, Y.; Wiig, J. A.
- Science, Vol. 331, Issue 6013
ATP-driven Reduction by Dark-operative Protochlorophyllide Oxidoreductase from Chlorobium tepidum Mechanistically Resembles Nitrogenase Catalysis
journal, February 2008
- Bröcker, Markus J.; Virus, Simone; Ganskow, Stefanie
- Journal of Biological Chemistry, Vol. 283, Issue 16
Structural analysis of cubane-type iron clusters
journal, July 2013
- Tan, Lay Ling; Holm, R. H.; Lee, Sonny C.
- Polyhedron, Vol. 58
Biosynthesis of the Metalloclusters of Nitrogenases
journal, June 2016
- Hu, Yilin; Ribbe, Markus W.
- Annual Review of Biochemistry, Vol. 85, Issue 1
Structural basis of biological nitrogen fixation
journal, April 2005
- Rees, Douglas C.; Akif Tezcan, F.; Haynes, Chad A.
- Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences, Vol. 363, Issue 1829
Nitrogenase Cofactor Assembly: An Elemental Inventory
journal, October 2017
- Sickerman, Nathaniel S.; Ribbe, Markus W.; Hu, Yilin
- Accounts of Chemical Research, Vol. 50, Issue 11
Radical SAM-Dependent Carbon Insertion into the Nitrogenase M-Cluster
journal, September 2012
- Wiig, J. A.; Hu, Y.; Lee, C. C.
- Science, Vol. 337, Issue 6102
Spectroscopic Characterization of an Eight‐Iron Nitrogenase Cofactor Precursor that Lacks the “9 th Sulfur”
journal, September 2019
- Jasniewski, Andrew J.; Wilcoxen, Jarett; Tanifuji, Kazuki
- Angewandte Chemie, Vol. 131, Issue 41
Evidence for Interstitial Carbon in Nitrogenase FeMo Cofactor
journal, November 2011
- Spatzal, T.; Aksoyoglu, M.; Zhang, L.
- Science, Vol. 334, Issue 6058
FeMo cofactor maturation on NifEN
journal, October 2006
- Hu, Y.; Corbett, M. C.; Fay, A. W.
- Proceedings of the National Academy of Sciences, Vol. 103, Issue 46
Genetic Transfer of Nitrogen Fixation from Klebsiella pneumoniae to Escherichia coli
journal, May 1972
- Dixon, R. A.; Postgate, J. R.
- Nature, Vol. 237, Issue 5350
Mechanism of Molybdenum Nitrogenase
journal, January 1996
- Burgess, Barbara K.; Lowe, David J.
- Chemical Reviews, Vol. 96, Issue 7
X-ray Emission Spectroscopy Evidences a Central Carbon in the Nitrogenase Iron-Molybdenum Cofactor
journal, November 2011
- Lancaster, K. M.; Roemelt, M.; Ettenhuber, P.
- Science, Vol. 334, Issue 6058
Transfer of Nitrogen-fixation Genes by Conjugation in Klebsiella pneumoniae
journal, November 1971
- Dixon, R. A.; Postgate, J. R.
- Nature, Vol. 234, Issue 5323
The biosynthetic pathway of coenzyme F430 in methanogenic and methanotrophic archaea
journal, October 2016
- Zheng, Kaiyuan; Ngo, Phong D.; Owens, Victoria L.
- Science, Vol. 354, Issue 6310
Assembly scaffold NifEN: A structural and functional homolog of the nitrogenase catalytic component
journal, August 2016
- Fay, Aaron W.; Blank, Michael A.; Rebelein, Johannes G.
- Proceedings of the National Academy of Sciences, Vol. 113, Issue 34
Fischer-Tropsch Chemistry at Room Temperature?
journal, July 2011
- Gerlach, Deidra L.; Lehnert, Nicolai
- Angewandte Chemie International Edition, Vol. 50, Issue 35
Tracing the ‘ninth sulfur’ of the nitrogenase cofactor via a semi-synthetic approach
journal, April 2018
- Tanifuji, Kazuki; Lee, Chi Chung; Sickerman, Nathaniel S.
- Nature Chemistry, Vol. 10, Issue 5
Identification and characterization of functional homologs of nitrogenase cofactor biosynthesis protein NifB from methanogens
journal, November 2015
- Fay, Aaron W.; Wiig, Jared A.; Lee, Chi Chung
- Proceedings of the National Academy of Sciences, Vol. 112, Issue 48
Characterization of Isolated Nitrogenase FeVco
journal, September 2010
- Fay, Aaron W.; Blank, Michael A.; Lee, Chi Chung
- Journal of the American Chemical Society, Vol. 132, Issue 36
Structural Models of the [Fe 4 S 4 ] Clusters of Homologous Nitrogenase Fe Proteins
journal, August 2011
- Blank, Michael A.; Lee, Chi Chung; Hu, Yilin
- Inorganic Chemistry, Vol. 50, Issue 15