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

doi:10.1002/1873-3468.13004

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

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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 language	English (US)
Pages (from-to)	987-998
Number of pages	12
Journal	FEBS Letters
Volume	592
Issue number	6
DOIs	https://doi.org/10.1002/1873-3468.13004
State	Published - Mar 2018

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Publisher Copyright:
© 2018 Federation of European Biochemical Societies

Keywords

OleA
X-ray crystallography
thiolase

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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.",

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AU - Esler, Morgan A.

AU - Christenson, James K

AU - Mohamed, Fatuma A.

AU - Wackett, Lawrence P

AU - Wilmot, Carrie M

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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.

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

Abstract

Bibliographical note

Keywords

UN SDGs

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