Professor of Chemistry and Biochemistry; BS in Chemistry, University of Redlands, California; PhD in Biochemistry, University of California, Riverside; Postdoctoral Fellow, Center for Metalloenzyme Studies, University of Georgia.
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Lance C. Seefeldt Professor of Chemistry and Biochemistry; BS in Chemistry, University of Redlands, California; PhD in Biochemistry, University of California, Riverside; Postdoctoral Fellow, Center for Metalloenzyme Studies, University of Georgia. | |
Contact Information Funding Publications Team |
Research Description Biochemistry of Nitrogen Fixation and Bioenergy Production The Seefeldt research program focuses on elucidating the mechanism of complex metalloenzymes involved in biological nitrogen fixation and global energy cycling. Nitrogenase is the enzyme responsible for biological reduction of dinitrogen from air into ammonia, catalyzing a critical reaction in the reductive phase of the global nitrogen cycle. Research is focused on answering the major open questions about nitrogenase mechanism including: 1) How is ATP hydrolysis coupled to substrate reduction; 2) How are electrons delivered to the active site metal clusters; and 3) where and how do substrates get reduced at the active site metal cluster FeMo-cofactor? The research approach utilizes genetics, biochemistry, spectroscopy including EPR and ENDOR, and X-ray crystallography.  A second research program is focused on exploring the use of phototrophic microbes as a way to capture light and carbon dioxide
to make feedstock for liquid transportation fuels. Projects
include isolation of robust microalgae from the Great Salt Lake that
can grow in salt water and that make high quantities of precursors for
fuels. Additionally, a suite of novel bacterial enzymes are being
studied that can be used to produce biodiesel in cells. | Recent News Sept. 2010 Dr. Yang paper to appear in Angewandte Chemie Sept. 2010 Danyal paper appears as a Communication in J. Am. Chem. Soc. Nov. 2010 Wahlen and Willis paper accepted in Bioresource Technology |
2009-2011 Uintah Basin Mitigation Project. “Carbon dioxide capture and water remediation”. (PI. Seefeldt).
2009-2011 Department of Energy. “Multifunctional Solar Energy Systems” (PI. Jeff Muhs, CoPIs Wood, Seefeldt, Sims).
2009-2012 National Science Foundation. “Bacterial Enzyme Systems for Wax Ester Production”. (PI. Brett Barney, Univ. of Minnesota, CoPI Seefeldt).
2009-2013 National Institutes of Health. “Nitrogenase Mechanism” (PI. Seefeldt, Co-PI
Dean).
2009-2012 Department of Energy. "Extremophilic Microalgae: Advanced
Lipid and Biomass Production for Biofuels and Bioproducts" (PI.
Brent Payton Montana State University; Co-PI Seefeldt, Barney, Viamajala).
1. Arp, D. J., McCollum. L. C., and Seefeldt, L. C. 1985. Molecular and immunological comparison of membrane-bound, H2-oxidizing hydrogenases of Bradyrhizobium japonicum, Alcaligenes eutrophus, Alcaligenes latus, and Azotobacter vinelandii. J. Bacteriol. 163, 15-20.
2. Seefeldt, L. C. and Arp, D. J. 1986. Purification to homogeneity of Azotobacter vinelandii hydrogenase: a nickel and iron containing dimer. Biochimie 68, 25-34.
3. Seefeldt, L. C., Fox, C. A., and Arp, D. J. 1986. Reversible inactivation of the O2-labile hydrogenases from Azotobacter vinelandii and Rhizobium japonicum. J. Biol. Chem. 261, 10688-10694.
4. Seefeldt, L. C., McCollum, L. C., Doyle, C. M., and Arp, D. J. 1987. Immunological and molecular evidence for a membrane-bound, dimeric hydrogenase in Rhodopseudomonas capsulata. Biochim. Biophys. Acta 914, 299-303.
5. Seefeldt, L. C. and Arp, D. J. 1987. Redox-dependent subunit dissociation of Azotobacter vinelandii hydrogenase in the presence of sodium dodecyl sulfate. J. Biol. Chem. 262, 16816-16821.
6. Hyman, M. R., Seefeldt, L. C., and Arp, D. J. 1988. Aerobic, inactive forms of Azotobacter vinelandii hydrogenase: activation kinetics and insensitivity to C2H2 inhibition. Biochim. Biophys. Acta 957, 91-96.
7. Kovacs, K. L., Seefeldt, L. C., Tigyi, G., Doyle, C. M., Mortenson, L. E., and Arp, D. J. 1989. Immunological relationship among hydrogenases. J. Bacteriol. 171, 430-435.
8. Seefeldt, L. C. and Arp, D. J. 1989. Oxygen effects on the nickel- and iron-containing hydrogenase from Azotobacter vinelandii. Biochemistry 28, 1588-1596.
9. Seefeldt, L. C. and Arp, D. J. 1989. Cyanide inactivation of hydrogenase from Azotobacter vinelandii. J. Bacteriol. 171, 3298-3303.
10. Ford, C. M., Garg, N., Garg, R. P., Tibelius, K. H., Yates, M. G., Arp, D. J., and Seefeldt, L. C. 1990. The identification, characterization, sequencing and mutagenesis of the genes (hupSL) encoding the small and large subunits of the H2-uptake hydrogenase of Azotobacter chroococcum. Molecular Microbiol. 4, 999-1008.
11. Hyman, M. R., Seefeldt, L. C., Morgan, T. V., Arp, D. J., and Mortenson, L. E. 1992. A kinetic and spectroscopic analysis of the inactivating effects of nitric oxide on the individual components of Azotobacter vinelandii nitrogenase. Biochemistry 31, 2947-2955.
12. Seefeldt,
L. C., Morgan, T. V., Dean, D. R., and
Mortenson, L. E. 1992. Mapping the site(s) of MgATP and MgADP interaction with
the nitrogenase of Azotobacter vinelandii:
Lysine 15 of the Fe protein plays a major role in MgATP interaction. J.
Biol. Chem. 267, 6680-6688.
13. Seefeldt, L.C. and Mortenson, L.E. 1993. Increasing nitrogenase catalytic efficiency for MgATP by site-directed mutagenesis of its Fe protein: Use of Mn++ to show interaction of serine 16 with Mg++. Protein Science 2, 93-102.
14. Seefeldt,
L.C. and
15. Seefeldt, L.C. 1994. Docking of nitrogenase iron- and molybdenum-iron proteins for electron transfer and MgATP hydrolysis: The role of arginine 140 and lysine 143 of the Azotobacter vinelandii iron protein. Protein Science 3, 2073-2081.
16. Seefeldt,
L.C.,
17. Ryle, M.J., Lanzilotta, W.N., Mortenson, L.E., Watt, G.D., and Seefeldt, L.C. 1995. Evidence for a central role of lysine 15 of Azotobacter vinelandii nitrogenase iron protein in nucleotide binding and protein conformational changes. J. Biol. Chem. 270, 13112-13117.
18. Lanzilotta, W.N., Ryle, M.J., and Seefeldt, L.C. 1995. Nucleotide hydrolysis and protein conformational changes in the Azotobacter vinelandii nitrogenase iron protein: Defining the function of aspartate 129. Biochemistry 34, 10713-10723.
19. Chen, J.C., Mortenson, L.E, and Seefeldt, L.C. 1995. Analysis of a gene region required for dihydrogen oxidation in Azotobacter vinelandii. Curr. Microbiol. 30, 351-355.
20. Lanzilotta,
W.N., Holz, R.C., and Seefeldt, L.C. 1995. Proton NMR investigation of the
[4Fe-4S]1+ cluster environment of
nitrogenase iron protein from Azotobacter
vinelandii: Defining nucleotide
induced conformational changes. Biochemistry
34, 15646-15653.
21. Ryle, M.J., Lanzilotta, W.N., Seefeldt, L.C., Scarrow, R.C., and Jensen, G.M. 1996. Circular dichroism and X-ray spectroscopies of Azotobacter vinelandii nitrogenase iron protein: MgATP and MgADP induced protein conformational changes affecting the [4Fe-4S] cluster and characterization of a [2Fe-2S] form. J. Biol. Chem. 271, 1551-1557.
22. Ryle, M.J. and Seefeldt, L.C. 1996. Elucidation of a MgATP signal transduction pathway in the nitrogenase iron protein: Formation of a conformation resembling the MgATP-bound state by protein engineering. Biochemistry 35, 4766-4775.
23. Lanzilotta, W.N., Fisher, K., and Seefeldt, L.C. 1996. Evidence for electron transfer from the nitrogenase iron protein to the molybdenum-iron protein without MgATP hydrolysis: Characterization of a tight protein-protein complex. Biochemistry 35, 7188-7196.
24. Ryle, M.J., Lanzilotta, W.N., and Seefeldt, L.C. 1996. Elucidating the mechanism of nucleotide-dependent changes in the redox potential of the [4Fe-4S] cluster in nitrogenase iron protein: The role of phenylalanine 135. Biochemistry 35, 9424-9434.
25. Ryle, M.J. and Seefeldt, L.C. 1996. The [4Fe-4S] cluster domain of the nitrogenase iron protein facilitates conformational changes required for the cooperative binding of two nucleotides. Biochemistry 35, 15654-15662.
26. Lanzilotta, W.N. and Seefeldt, L.C. 1996. Electron transfer from the nitrogenase iron protein to the [8Fe-(7/8)] clusters of the molybdenum-iron protein. Biochemistry 35, 16770-16776.
27. Lanzilotta, W.N., Fisher, K., and Seefeldt, L.C. 1997. Evidence for electron transfer-dependent formation of a nitrogenase iron protein-molybdenum-iron protein tight complex: The role of aspartate 39. J. Biol. Chem. 272, 4157-4165.
28. Seefeldt, L.C. and Dean, D.R. 1997. Role of nucleotides in nitrogenase catalysis. Acc. Chem. Res. 30, 260-266.
29. Parker, V.D. and Seefeldt, L.C. 1997. A mediated thin-layer voltammetry method for the study of redox protein electrochemistry. Anal. Biochem. 247, 152-157.
30.
31. Parker, V.D., Roddick, A., Seefeldt, L.C., Wang, H., and Zheng, G. 1997. Determination of rate and equilibrium constants for the reactions between electron transfer mediators and proteins by linear sweep voltammetry. Anal. Biochem. 249, 212-218.
32. Lanzilotta, W.N. and Seefeldt, L.C. 1997. Changes in the midpoint potentials of the nitrogenase metal centers as a result of iron protein-molybdenum-iron protein complex formation. Biochemistry 36, 12976-12983.
33. Liu, Y., Seefeldt, L.C., and Parker, V.D. 1997. Entropies of redox reactions between proteins and mediators: the temperature dependence of reversible electrode potentials in aqueous buffers. Anal. Biochem. 250, 196-202.
34. Lanzilotta, W.N., Parker, V.D., and Seefeldt, L.C. 1998. Electron transfer in nitrogenase analyzed by Marcus theory: Evidence for gating by MgATP. Biochemistry 37, 399-407.
35. Lanzilotta, W.N., Christiansen, J., Dean, D.R., and Seefeldt, L.C. 1998. Evidence for coupled electron and proton transfer in the [8Fe-7S] cluster of nitrogenase. Biochemistry 37, 11376-11384.
36. Christiansen, J., Goodwin, P.J., Lanzilotta, W.N., Seefeldt, L.C., and Dean, D.R. 1998. Catalytic and biophysical properties of a nitrogenase apo-MoFe protein produced by a nifB-deletion mutant of Azotobacter vinelandii. Biochemistry 37, 12611-12623.
37. Peters, J.W., Lanzilotta, W.N., Lemon, B.J., and Seefeldt, L.C. 1998. The x-ray crystal structure of the Fe-only hydrogenase (CpI) from Clostridium pasteurianum Science 282, 1853-1858. to 1.8 angstrom resolution.
38. Lanzilotta, W.N., Parker, V.D., and Seefeldt, L.C. 1999. Thermodynamics of nucleotide interactions with the Azotobacter vinelandii nitrogenase iron protein. Biochim. Biophys. Acta 1429, 411-421.
39. Chan, J.M., Christiansen, J., Dean, D.R., and Seefeldt, L.C. 1999. Spectroscopic evidence for changes in the redox state of the nitrogenase P-cluster during turnover. Biochemistry 38, 5779-5785.
40. Chan, J.M., Ryle, M.J., and Seefeldt, L.C. 1999. Evidence that MgATP accelerates primary electron transfer in a Clostridium pasteurianum Fe protein-Azotobacter vinelandii MoFe protein nitrogenase tight complex. J. Biol. Chem. 274, 17593-17598.
41. Jang, S.B., Seefeldt, L.C., and Peters, J.W. 2000. Modulating the midpoint potential of the [4Fe-4S] cluster of the nitrogenase Fe protein. Biochemistry 39, 641-648.
42. Christiansen, J., Chan, J.M., Seefeldt, L.C., and Dean, D.R. 2000. The role of the MoFe protein a-125Phe and b-125Phe residues in Azotobacter vinelandii J. Inorg. Biochem. 80, 195-204. MoFe protein-Fe protein interactions.
43. Christiansen, J., Cash, V.L., Seefeldt, L.C., Dean, D.R. 2000. Isolationi and characterization of an acetylene resistant nitrogenase. J. Biol. Chem. 275, 11459-11464.
44. Ryle, M.J., Lee, H.I., Seefeldt, L.C., and Hoffman, B.M. 2000. Nitrogenase reduction of carbon disulfide: freeze quench EPR and ENDOR evidence for three sequential intermediates with cluster-bound carbon moieties. Biochemistry 39, 1114-1119.
45. Ryle, M.J., and Seefeldt, L.C. 2000. Hydrolysis of nucleoside triphosphates other than ATP by nitrogenase. J. Biol. Chem. 275, 6214-6219.
46. Chan, J.M., Wu, W., Dean, D.R., and Seefeldt, L.C. 2000. Construction and characterization of a heterodimeric Fe protein: Defining roles for ATP in nitrogenase catalysis. Biochemistry 39, 7221-7228.
47. Christiansen, J. Seefeldt, L.C., and Dean, D.R. 2000. Competitive substrate and inhibitor interactions at the physiologically relevant active site of nitrogenase. J. Biol. Chem. 275, 36104-36107.
48. Jang, S.B., Seefeldt, L.C., and Peters, J.W. 2000. Insights into nucleotide signal transduction in nitrogenase: structure of an iron protein with MgADP bound. Biochemistry 39, 14745-14752.
49. Sorlie, M., Seefeldt, L.C., and Parker, V.D. 2000. Use of stopped-flow spectrophotometry to establish midpoint potentials for redox proteins. Anal. Biochem.287, 118-125.
50. Chiu,
H.J., Peters, J.W., Lanzilotta, W.N., Ryle, M.J., Seefeldt, L.C., Howard, J.B.,
and
51.
52.
53. Sorlie, M., Chan, J.M., Wang, H., Seefeldt, L.C., and Parker, V.D. 2003. Elucidating thermodynamic parameters for electron transfer proteins using isothermal titration calorimetry: application to the nitrogenase Fe protein. J. Biol. Inorg. Chem. 8, 560-566.
54.
Lee, H.I.,
55.
56. Igarashi, R.Y. and Seefeldt, L.C. 2003. Nitrogen fixation: the mechanism of the Mo-dependent nitrogenase. Crit. Rev. Biochem. Mol. Biol. 38, 351-384.
57.
Seefeldt, L.C., Dance,
58. Sen, S., Igarashi, R., Smith, A. Johnson, M.K., Seefeldt, L.C., and Peters, J.W. 2004. A conformational mimic of the MgATP-bound “on state” of the nitrogenase Fe protein. Biochemistry 43, 1787-1797.
59.
Igarashi, R.Y., Dos
60.
Lee, H.I., Igarashi, R.Y., Laryukhin, M., Doan, P.E., Dos
61.
Barney, B.M., Igarashi, R.Y., Dos
62. Jang, S.B., Jeong, M.S., Seefeldt, L.C., Peters, J.W. 2004. Structural and biochemical implications of single amino acid substitutions in the nucleotide-dependent switch regions of the nitrogenase Fe protein from Azotobacter vinelandii. J. Biol. Inorg. Chem. 9, 1028-1033.
63.
Dos
64.
Igarashi, R.Y., Laryukhin, M., Dos
65.
Barney, B.M., Laryukhin, M., Igarashi, R.Y., Lee, H.I., Dos
66. Yang, T.C., Maeser, N.K., Laryukhin, M., Lee, H.I., Dean, D.R., Seefeldt, L.C., and Hoffman, B.M. 2005. The interstitial atom of the nitrogenase FeMo-cofactor: ENDOR and ESEEM evidence that it is not a nitrogen. J. Am. Chem. Soc. 127, 12804-12805.
67.
Barney, B.M., Yang, T.C., Igarashi, R.Y., Dos
68.
Barney, B.M., Lee, H.I., Dos
69. Sen, S., Krishnakumar, A., McClead, J., Johnson, M.K., Seefeldt, L.C., Szilagyi, R.K., and Peters, J.W. 2006. Insights into the role of nucletotide-dependent conformational change in nitrogenase catalysis: structural characterization of the nitrogenase Fe protein Leu127 deletion variant with bound MgATP. J. Inorg. Biochem. 100, 1041-1052.
70. Barney, B.M., Lukoyanov, D., Yang, T.C., Dean, D.R., Hoffman, B.M., and Seefeldt, L.C. 2006. A methyldiazene (HN=N-CH3)-derived species bound to the nitrogenase active site FeMo-cofactor: implications for mechanism. Proc. Nat. Acad. Sci. USA 103, 17113-17118.
71.
Lukoyanov, D., Barney, BM., Dean, D.R., Seefeldt, L.C., and Hoffman, B.M. 2007.
Connecting nitrogenase intermediates with the kinetic scheme for N2
reduction by a relaxation protocol and identification of the N2
binding state. Proc. Nat. Acad. Sci. USA 104, 1451-1455.
72. Barney, B.M., McClead, J., Lukoyanov, D., Laryukhin, M., Yang, T.C., Dean, D.R., Hoffman, B.M., Seefeldt, L.C. 2007. Diazene (HN=NH) is a substrate for nitrogenase: insights into the pathway of N2 reduction. Biochemistry 46, 6784-6794.
73. Dos
74.
Sarma, R., Mulder, D., Brecht, E., Szilagyi, R., Seefeldt, L.C., Tsuruta, H.,
Peters, J.W. (2007) Probing the MgATP-bound conformation of the nitrogenase Fe
protein by solution small angle x-ray scattering. Biochemistry 46, 14058-14066.
75.
Lukoyanov, D., V. Pelmenschikov, Maeser, N., Laryukhin, M., Yang, T.C.,
Noodleman, L., Dean, D.R., Case, D.A., Seefeldt, L.C., and Hoffman, B.M. (2007)
Testing if the interstitial atom, X, of the nitrogenase molybdenum-iron
cofactor is N or C: ENDOR, ESEEM, and DFT studies of the S = 3/2 resting state
in multiple environments. Inorg.
Chem. 46, 11437-11449.
76.
Wahlen, B.D., Barney, B.M., and Seefeldt, L.C. (2008) Synthesis of biodiesel from mixed feedstocks
and longer chain alcohols using an acid catalyzed method. Energy Fuels 22, 4223-4228.
77. Sarma, R., Barney, B.M.,
78. Seefeldt, L.C., Hoffman,
B.M., Dean, D.R. (2009) Mechanism of Mo-dependent nitrogenase. Ann.
Rev. Biochem. 78, 701-722.
79. Hoffman, B.M., Dean, D.R.,
Seefeldt, L.C. (2009) Climbing nitrogenase: towards a mechanism of enzymatic
nitrogen fixation. Acc. Chem. Res. 42, 609-619.
80. Wahlen, B.D., Oswald, W.S.,
Seefeldt, L.C., Barney, B.M. (2009) Purification, characterization, and
potential bacterial wax ester production role of an NADPH-dependent fatty
aldehyde reductase from Marinobacter
aquaeolei VT8. Appl. Environ. Microbiol. 75, 2758-2764.
81. Barney, B.M., Yurth, M.G.,
Dos Santos, P.C., Dean, D.R., Seefeldt, L.C. (2009) A substrate channel in the
nitrogenase MoFe protein. J.
Biol. Inorg. Chem. 14, 1015-1022.
82. Barney, B.M., Lukoyanov, D.,
Igarashi, R.Y., Laryukhin, M., Yang, T.C., Dean, D.R., Hoffman, B.M., Seefeldt,
L.C. (2009). Trapping an intermediate of
dinitrogen (N2) reduction on nitrogenase.
Biochemistry 48, 9094-9102.
83. Sarma, R., Barney, B.M.,
Keable, S., Dean, D.R., Seefeldt, L.C., and Peters, J.W. (2010). Insights into substrate binding at
FeMo-cofactor in nitrogenase from the structure of an alpha70Ile MoFe protein
variant. J. Inorg. Biochem. 104,
385-389.
84. Lukoyanov, D., Yang, Z.Y.,
Dean, D.R., Seefeldt, L.C., and Hoffman, B.M. (2010). Is Mo involved in hydride
binding by the four-electron reduced (E4) intermediate of the nitrogenase MoFe
protein? J. Am. Chem. Soc. 132, 2526-2527.
85. Danyal, K., Mayweather, D.,
Dean, D.R., Seefeldt, L.C., and Hoffman, B.M. (2010) Conformational gating of
electron transfer from nitrogenase Fe protein to MoFe protein. J. Am. Chem. Soc. 132,
6894-6895.
86. Danyal, K., Inglet, B.S., Vincent, K.A., Barney, B.M., Hoffman, B.M., Armstrong, F.A, Dean, D.R., and Seefeldt, L.C. Uncoupling nitrogenase: catalytic reduction of hydrazine to ammonia by a MoFe protein in the absence of Fe protein-ATP J. Am. Chem. Soc. (in press).
87. Yang, Z.Y., Seefeldt, L.C.,
Dean, D.R., Cramer, S.P., and George, S.J. Steric control of the Hi-CO MoFe
nitrogenase complex revealed by stopped-flow infra-red spectroscopy. Ang. Chem. Int. Ed. (in press).
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brad.wahlen@aggiemail.usu.edu |
 K. Danyal Graduate Student |
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| Brett Barney | Postdoctoral Fellow 2009 | Assistant Professor, Biological Engineering, University of Minnesota |
| Madeline Rasche | Postdoctoral Fellow 1997 | Professor, Biochemistry Department, California State University Fullerton |
| Jenny Huyett | Postdoctoral Fellow 2000 | Research Scientist, Boulder CO |
| Paul Benton | Postdoctoral Fellow 2002 | Chemistry Department, Montana State University |
| William Lanzilotta |
PhD 1999 | Associate Professor, Biochemistry Department, University of Georgia |
| Matt Ryle |
PhD 1998 | Research Scientist, IDEXX Laboratories, Inc., Westbrook, ME |
| Morten Sorlie |
PhD 2000 |
Professor of Chemistry, Norwegian University of Life Sciences, As, Norway |
| Jeannine Chan |
PhD 2001 | Assistant Professor, Pacific University |
| Robert Igarashi |
PhD 2005 | Assistant Professor, Chemistry Department, University of Central Florida |
| Wei Wu |
MS 2001 | Nutrition and Food Science, Utath State University |
| Jammi McClead |
MS 2007 | Ohio State University |
| Alejandro Pabon |
MS 2008 |
Utah Valley University |
| Nate Maeser |
MS 2008 |
Medical School Australia |
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BS Chemistry |
MD Johns Hopkins Univ. |
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Fort Lewis College, CO |
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MD Virginia Commonwealth |
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