Different Behaviors of a Substrate in P450 Decarboxylase and Hydroxylase Reveal Reactivity-Enabling Actors
Abstract
Biological routes to the production of fuels from renewable feedstocks hold significant promise in our efforts towards a sustainable future. The fatty acid decarboxylase enzyme (OleTJE) is a cytochrome P450 enzyme that converts long and medium chain fatty acids to terminal alkenes and shares significant similarities in terms of structure, substrate scope and mechanism with the hydroxylase cytochrome P450 (P450BSβ). Recent reports have demonstrated that catalytic pathways in these enzymes bifurcate when the heme is in its iron-hydroxo (compound II) state. In spite of significant similarities, the fundamental underpinnings of their different characteristic wild-type reactivities remain ambiguous. In this work, we develop point charges, modified parameters and report molecular simulations of this crucial intermediate step. Water occupancies and substrate mobility at the active site are observed to be vital differentiating aspects between the two enzymes in the compound II state and corroborate recent experimental hypotheses. Apart from increased substrate mobility in the hydroxylase, which could have implications for enabling the rebound mechanism for hydroxylation, OleTJE is characterized by much stronger binding of the substrate carboxylate group to the active site arginine, implicating it as an important enabling actor for decarboxylation.
- Authors:
-
[1];
[1];
[2];
- National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center
- National Renewable Energy Lab. (NREL), Golden, CO (United States). National Bioenergy Center
- Publication Date:
- Research Org.:
- National Renewable Energy Laboratory (NREL), Golden, CO (United States)
- Sponsoring Org.:
- USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office (BETO); USDOE Laboratory Directed Research and Development (LDRD) Program
- OSTI Identifier:
- 1470966
- Report Number(s):
- NREL/JA-2700-72223
Journal ID: ISSN 2045-2322
- Grant/Contract Number:
- AC36-08GO28308
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Scientific Reports
- Additional Journal Information:
- Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 2045-2322
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; P450s; compound II; decarboxylase; OleTJE; hydroxylase; hydrocarbon bioproduction; molecular dynamics; heme; water occupancy
Citation Formats
Bharadwaj, Vivek S., Kim, Seonah, Guarnieri, Michael T., and Crowley, Michael F. Different Behaviors of a Substrate in P450 Decarboxylase and Hydroxylase Reveal Reactivity-Enabling Actors. United States: N. p., 2018.
Web. doi:10.1038/s41598-018-31237-4.
Bharadwaj, Vivek S., Kim, Seonah, Guarnieri, Michael T., & Crowley, Michael F. Different Behaviors of a Substrate in P450 Decarboxylase and Hydroxylase Reveal Reactivity-Enabling Actors. United States. https://doi.org/10.1038/s41598-018-31237-4
Bharadwaj, Vivek S., Kim, Seonah, Guarnieri, Michael T., and Crowley, Michael F. Mon .
"Different Behaviors of a Substrate in P450 Decarboxylase and Hydroxylase Reveal Reactivity-Enabling Actors". United States. https://doi.org/10.1038/s41598-018-31237-4. https://www.osti.gov/servlets/purl/1470966.
@article{osti_1470966,
title = {Different Behaviors of a Substrate in P450 Decarboxylase and Hydroxylase Reveal Reactivity-Enabling Actors},
author = {Bharadwaj, Vivek S. and Kim, Seonah and Guarnieri, Michael T. and Crowley, Michael F.},
abstractNote = {Biological routes to the production of fuels from renewable feedstocks hold significant promise in our efforts towards a sustainable future. The fatty acid decarboxylase enzyme (OleTJE) is a cytochrome P450 enzyme that converts long and medium chain fatty acids to terminal alkenes and shares significant similarities in terms of structure, substrate scope and mechanism with the hydroxylase cytochrome P450 (P450BSβ). Recent reports have demonstrated that catalytic pathways in these enzymes bifurcate when the heme is in its iron-hydroxo (compound II) state. In spite of significant similarities, the fundamental underpinnings of their different characteristic wild-type reactivities remain ambiguous. In this work, we develop point charges, modified parameters and report molecular simulations of this crucial intermediate step. Water occupancies and substrate mobility at the active site are observed to be vital differentiating aspects between the two enzymes in the compound II state and corroborate recent experimental hypotheses. Apart from increased substrate mobility in the hydroxylase, which could have implications for enabling the rebound mechanism for hydroxylation, OleTJE is characterized by much stronger binding of the substrate carboxylate group to the active site arginine, implicating it as an important enabling actor for decarboxylation.},
doi = {10.1038/s41598-018-31237-4},
journal = {Scientific Reports},
number = 1,
volume = 8,
place = {United States},
year = {Mon Aug 27 00:00:00 EDT 2018},
month = {Mon Aug 27 00:00:00 EDT 2018}
}
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Works referencing / citing this record:
Directed Evolution of P450 Fatty Acid Decarboxylases via High‐Throughput Screening towards Improved Catalytic Activity
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