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

1 Scopus citations

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 2018

Keywords

  • OleA
  • X-ray crystallography
  • thiolase

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    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