The role of OleA His285 in orchestration of long-chain acyl-coenzyme A substrates

Matthew R Jensen, Brandon R. Goblirsch, Morgan A. Esler, James K Christenson, Fatuma A. Mohamed, Lawrence P Wackett, Carrie M Wilmot

Research output: Contribution to journalLetter

Abstract

Renewable production of hydrocarbons is being pursued as a petroleum-independent source of commodity chemicals and replacement for biofuels. The bacterial biosynthesis of long-chain olefins represents one such platform. The process is initiated by OleA catalyzing the condensation of two fatty acyl-coenzyme A substrates to form a β-keto acid. Here, the mechanistic role of the conserved His285 is investigated through mutagenesis, activity assays, and X-ray crystallography. Our data demonstrate that His285 is required for product formation, influences the thiolase nucleophile Cys143 and the acyl-enzyme intermediate before and after transesterification, and orchestrates substrate coordination as a defining component of an oxyanion hole. As a consequence, His285 plays a key role in enabling a mechanistic strategy in OleA that is distinct from other thiolases.

Original languageEnglish (US)
Pages (from-to)987-998
Number of pages12
JournalFEBS Letters
Volume592
Issue number6
DOIs
StatePublished - Mar 1 2018

Fingerprint

Acyl Coenzyme A
Olea
Keto Acids
Mutagenesis
Nucleophiles
Biofuels
X ray crystallography
X Ray Crystallography
Biosynthesis
Transesterification
Petroleum
Alkenes
Substrates
Hydrocarbons
Condensation
Assays
Enzymes

Keywords

  • OleA
  • X-ray crystallography
  • thiolase

PubMed: MeSH publication types

  • Letter
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

Cite this

Jensen, M. R., Goblirsch, B. R., Esler, M. A., Christenson, J. K., Mohamed, F. A., Wackett, L. P., & Wilmot, C. M. (2018). The role of OleA His285 in orchestration of long-chain acyl-coenzyme A substrates. FEBS Letters, 592(6), 987-998. https://doi.org/10.1002/1873-3468.13004

The role of OleA His285 in orchestration of long-chain acyl-coenzyme A substrates. / Jensen, Matthew R; Goblirsch, Brandon R.; Esler, Morgan A.; Christenson, James K; Mohamed, Fatuma A.; Wackett, Lawrence P; Wilmot, Carrie M.

In: FEBS Letters, Vol. 592, No. 6, 01.03.2018, p. 987-998.

Research output: Contribution to journalLetter

Jensen, MR, Goblirsch, BR, Esler, MA, Christenson, JK, Mohamed, FA, Wackett, LP & Wilmot, CM 2018, 'The role of OleA His285 in orchestration of long-chain acyl-coenzyme A substrates', FEBS Letters, vol. 592, no. 6, pp. 987-998. https://doi.org/10.1002/1873-3468.13004
Jensen MR, Goblirsch BR, Esler MA, Christenson JK, Mohamed FA, Wackett LP et al. The role of OleA His285 in orchestration of long-chain acyl-coenzyme A substrates. FEBS Letters. 2018 Mar 1;592(6):987-998. https://doi.org/10.1002/1873-3468.13004
Jensen, Matthew R ; Goblirsch, Brandon R. ; Esler, Morgan A. ; Christenson, James K ; Mohamed, Fatuma A. ; Wackett, Lawrence P ; Wilmot, Carrie M. / The role of OleA His285 in orchestration of long-chain acyl-coenzyme A substrates. In: FEBS Letters. 2018 ; Vol. 592, No. 6. pp. 987-998.
@article{f14d0058405341699c1f57aebadadcd6,
title = "The role of OleA His285 in orchestration of long-chain acyl-coenzyme A substrates",
abstract = "Renewable production of hydrocarbons is being pursued as a petroleum-independent source of commodity chemicals and replacement for biofuels. The bacterial biosynthesis of long-chain olefins represents one such platform. The process is initiated by OleA catalyzing the condensation of two fatty acyl-coenzyme A substrates to form a β-keto acid. Here, the mechanistic role of the conserved His285 is investigated through mutagenesis, activity assays, and X-ray crystallography. Our data demonstrate that His285 is required for product formation, influences the thiolase nucleophile Cys143 and the acyl-enzyme intermediate before and after transesterification, and orchestrates substrate coordination as a defining component of an oxyanion hole. As a consequence, His285 plays a key role in enabling a mechanistic strategy in OleA that is distinct from other thiolases.",
keywords = "OleA, X-ray crystallography, thiolase",
author = "Jensen, {Matthew R} and Goblirsch, {Brandon R.} and Esler, {Morgan A.} and Christenson, {James K} and Mohamed, {Fatuma A.} and Wackett, {Lawrence P} and Wilmot, {Carrie M}",
year = "2018",
month = "3",
day = "1",
doi = "10.1002/1873-3468.13004",
language = "English (US)",
volume = "592",
pages = "987--998",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "6",

}

TY - JOUR

T1 - The role of OleA His285 in orchestration of long-chain acyl-coenzyme A substrates

AU - Jensen, Matthew R

AU - Goblirsch, Brandon R.

AU - Esler, Morgan A.

AU - Christenson, James K

AU - Mohamed, Fatuma A.

AU - Wackett, Lawrence P

AU - Wilmot, Carrie M

PY - 2018/3/1

Y1 - 2018/3/1

N2 - Renewable production of hydrocarbons is being pursued as a petroleum-independent source of commodity chemicals and replacement for biofuels. The bacterial biosynthesis of long-chain olefins represents one such platform. The process is initiated by OleA catalyzing the condensation of two fatty acyl-coenzyme A substrates to form a β-keto acid. Here, the mechanistic role of the conserved His285 is investigated through mutagenesis, activity assays, and X-ray crystallography. Our data demonstrate that His285 is required for product formation, influences the thiolase nucleophile Cys143 and the acyl-enzyme intermediate before and after transesterification, and orchestrates substrate coordination as a defining component of an oxyanion hole. As a consequence, His285 plays a key role in enabling a mechanistic strategy in OleA that is distinct from other thiolases.

AB - Renewable production of hydrocarbons is being pursued as a petroleum-independent source of commodity chemicals and replacement for biofuels. The bacterial biosynthesis of long-chain olefins represents one such platform. The process is initiated by OleA catalyzing the condensation of two fatty acyl-coenzyme A substrates to form a β-keto acid. Here, the mechanistic role of the conserved His285 is investigated through mutagenesis, activity assays, and X-ray crystallography. Our data demonstrate that His285 is required for product formation, influences the thiolase nucleophile Cys143 and the acyl-enzyme intermediate before and after transesterification, and orchestrates substrate coordination as a defining component of an oxyanion hole. As a consequence, His285 plays a key role in enabling a mechanistic strategy in OleA that is distinct from other thiolases.

KW - OleA

KW - X-ray crystallography

KW - thiolase

UR - http://www.scopus.com/inward/record.url?scp=85042133799&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85042133799&partnerID=8YFLogxK

U2 - 10.1002/1873-3468.13004

DO - 10.1002/1873-3468.13004

M3 - Letter

VL - 592

SP - 987

EP - 998

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 6

ER -