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Title: A promiscuous cytochrome P450 aromatic O-demethylase for lignin bioconversion

Microbial aromatic catabolism offers a promising approach to convert lignin, a vast source of renewable carbon, into useful products. Aryl-O-demethylation is an essential biochemical reaction to ultimately catabolize coniferyl and sinapyl lignin-derived aromatic compounds, and is often a key bottleneck for both native and engineered bioconversion pathways. Here, we report the comprehensive characterization of a promiscuous P450 aryl-O-demethylase, consisting of a cytochrome P450 protein from the family CYP255A (GcoA) and a three-domain reductase (GcoB) that together represent a new two-component P450 class. Though originally described as converting guaiacol to catechol, we show that this system efficiently demethylates both guaiacol and an unexpectedly wide variety of lignin-relevant monomers. Here, structural, biochemical, and computational studies of this novel two-component system elucidate the mechanism of its broad substrate specificity, presenting it as a new tool for a critical step in biological lignin conversion.
Authors:
 [1] ; ORCiD logo [2] ;  [3] ; ORCiD logo [4] ;  [5] ; ORCiD logo [6] ;  [1] ; ORCiD logo [7] ; ORCiD logo [6] ; ORCiD logo [8] ;  [4] ; ORCiD logo [6] ;  [9] ; ORCiD logo [1]
  1. Univ. of Portsmouth, Portsmouth (United Kingdom)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States); Montana State Univ., Bozeman, MT (United States)
  3. National Renewable Energy Lab. (NREL), Golden, CO (United States); Univ. of Campinas, Sao Paulo (Brazil)
  4. Univ. of California at Los Angeles, Los Angeles, CA (United States)
  5. National Renewable Energy Lab. (NREL), Golden, CO (United States); Univ. of California at Los Angeles, Los Angeles, CA (United States)
  6. National Renewable Energy Lab. (NREL), Golden, CO (United States)
  7. Univ. of Campinas, Sao Paulo (Brazil)
  8. Univ. of Georgia, Athens, GA (United States)
  9. Montana State Univ., Bozeman, MT (United States)
Publication Date:
Report Number(s):
NREL/JA-5100-71206
Journal ID: ISSN 2041-1723
Grant/Contract Number:
AC36-08GO28308
Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Research Org:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Bioenergy Technologies Office (EE-3B)
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; enzyme discovery; protein family; enzyme characterization; lignin conversion
OSTI Identifier:
1458818

Mallinson, Sam J. B., Machovina, Melodie M., Silveira, Rodrigo L., Garcia-Borras, Marc, Gallup, Nathan, Johnson, Christopher W., Allen, Mark D., Skaf, Munir S., Crowley, Michael F., Neidle, Ellen L., Houk, Kendall N., Beckham, Gregg T., DuBois, Jennifer L., and McGeehan, John E.. A promiscuous cytochrome P450 aromatic O-demethylase for lignin bioconversion. United States: N. p., Web. doi:10.1038/s41467-018-04878-2.
Mallinson, Sam J. B., Machovina, Melodie M., Silveira, Rodrigo L., Garcia-Borras, Marc, Gallup, Nathan, Johnson, Christopher W., Allen, Mark D., Skaf, Munir S., Crowley, Michael F., Neidle, Ellen L., Houk, Kendall N., Beckham, Gregg T., DuBois, Jennifer L., & McGeehan, John E.. A promiscuous cytochrome P450 aromatic O-demethylase for lignin bioconversion. United States. doi:10.1038/s41467-018-04878-2.
Mallinson, Sam J. B., Machovina, Melodie M., Silveira, Rodrigo L., Garcia-Borras, Marc, Gallup, Nathan, Johnson, Christopher W., Allen, Mark D., Skaf, Munir S., Crowley, Michael F., Neidle, Ellen L., Houk, Kendall N., Beckham, Gregg T., DuBois, Jennifer L., and McGeehan, John E.. 2018. "A promiscuous cytochrome P450 aromatic O-demethylase for lignin bioconversion". United States. doi:10.1038/s41467-018-04878-2. https://www.osti.gov/servlets/purl/1458818.
@article{osti_1458818,
title = {A promiscuous cytochrome P450 aromatic O-demethylase for lignin bioconversion},
author = {Mallinson, Sam J. B. and Machovina, Melodie M. and Silveira, Rodrigo L. and Garcia-Borras, Marc and Gallup, Nathan and Johnson, Christopher W. and Allen, Mark D. and Skaf, Munir S. and Crowley, Michael F. and Neidle, Ellen L. and Houk, Kendall N. and Beckham, Gregg T. and DuBois, Jennifer L. and McGeehan, John E.},
abstractNote = {Microbial aromatic catabolism offers a promising approach to convert lignin, a vast source of renewable carbon, into useful products. Aryl-O-demethylation is an essential biochemical reaction to ultimately catabolize coniferyl and sinapyl lignin-derived aromatic compounds, and is often a key bottleneck for both native and engineered bioconversion pathways. Here, we report the comprehensive characterization of a promiscuous P450 aryl-O-demethylase, consisting of a cytochrome P450 protein from the family CYP255A (GcoA) and a three-domain reductase (GcoB) that together represent a new two-component P450 class. Though originally described as converting guaiacol to catechol, we show that this system efficiently demethylates both guaiacol and an unexpectedly wide variety of lignin-relevant monomers. Here, structural, biochemical, and computational studies of this novel two-component system elucidate the mechanism of its broad substrate specificity, presenting it as a new tool for a critical step in biological lignin conversion.},
doi = {10.1038/s41467-018-04878-2},
journal = {Nature Communications},
number = 1,
volume = 9,
place = {United States},
year = {2018},
month = {6}
}

Works referenced in this record:

Lignin Biosynthesis
journal, June 2003

Electron spin relaxation of iron-sulphur proteins studied by microwave power saturation
journal, December 1978
  • Rupp, H.; Rao, K. K.; Hall, D. O.
  • Biochimica et Biophysica Acta (BBA) - Protein Structure, Vol. 537, Issue 2, p. 255-269
  • DOI: 10.1016/0005-2795(78)90509-3

Scalable molecular dynamics with NAMD
journal, January 2005
  • Phillips, James C.; Braun, Rosemary; Wang, Wei
  • Journal of Computational Chemistry, Vol. 26, Issue 16, p. 1781-1802
  • DOI: 10.1002/jcc.20289

Structure and function of cytochromes P450:a comparative analysis of three crystal structures
journal, January 1995

Lignin valorization through integrated biological funneling and chemical catalysis
journal, August 2014
  • Linger, J. G.; Vardon, D. R.; Guarnieri, M. T.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 33, p. 12013-12018
  • DOI: 10.1073/pnas.1410657111

Lignin Valorization: Improving Lignin Processing in the Biorefinery
journal, May 2014
  • Ragauskas, A. J.; Beckham, G. T.; Biddy, M. J.
  • Science, Vol. 344, Issue 6185, p. 1246843-1246843
  • DOI: 10.1126/science.1246843

Density‐functional thermochemistry. III. The role of exact exchange
journal, April 1993
  • Becke, Axel D.
  • The Journal of Chemical Physics, Vol. 98, Issue 7, p. 5648-5652
  • DOI: 10.1063/1.464913

Microbial degradation of aromatic compounds — from one strategy to four
journal, October 2011
  • Fuchs, Georg; Boll, Matthias; Heider, Johann
  • Nature Reviews Microbiology, Vol. 9, Issue 11, p. 803-816
  • DOI: 10.1038/nrmicro2652

PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010
  • Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221
  • DOI: 10.1107/S0907444909052925

EasySpin, a comprehensive software package for spectral simulation and analysis in EPR
journal, January 2006

A Tetrahydrofolate-Dependent O-Demethylase, LigM, Is Crucial for Catabolism of Vanillate and Syringate in Sphingomonas paucimobilis SYK-6
journal, March 2005