Substrate-triggered addition of dioxygen to the diferrous cofactor of aldehyde-deformylating oxygenase to form a diferric-peroxide intermediate

Maria E. Pandelia, Ning Li, Hanne Nørgaard, Douglas Warui, Lauren J. Rajakovich, Wei Chen Chang, Squire J. Booker, Carsten Krebs, Joseph M. Bollinger, Jr.

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Cyanobacterial aldehyde-deformylating oxygenases (ADOs) belong to the ferritin-like diiron-carboxylate superfamily of dioxygen-activating proteins. They catalyze conversion of saturated or monounsaturated Cn fatty aldehydes to formate and the corresponding Cn-1 alkanes or alkenes, respectively. This unusual, apparently redox-neutral transformation actually requires four electrons per turnover to reduce the O2 cosubstrate to the oxidation state of water and incorporates one O-atom from O2 into the formate coproduct. We show here that the complex of the diiron(II/II) form of ADO from Nostoc punctiforme (Np) with an aldehyde substrate reacts with O2 to form a colored intermediate with spectroscopic properties suggestive of a Fe2III/III complex with a bound peroxide. Its Mössbauer spectra reveal that the intermediate possesses an antiferromagnetically (AF) coupled Fe2III/III center with resolved subsites. The intermediate is long-lived in the absence of a reducing system, decaying slowly (t1/2 ∼ 400 s at 5 C) to produce a very modest yield of formate (<0.15 enzyme equivalents), but reacts rapidly with the fully reduced form of 1-methoxy-5-methylphenazinium methylsulfate ( MeOPMS) to yield product, albeit at only ∼50% of the maximum theoretical yield (owing to competition from one or more unproductive pathway). The results represent the most definitive evidence to date that ADO can use a diiron cofactor (rather than a homo- or heterodinuclear cluster involving another transition metal) and provide support for a mechanism involving attack on the carbonyl of the bound substrate by the reduced O2 moiety to form a Fe2III/III-peroxyhemiacetal complex, which undergoes reductive O-O-bond cleavage, leading to C1-C2 radical fragmentation and formation of the alk(a/e)ne and formate products.

Original languageEnglish (US)
Pages (from-to)15801-15812
Number of pages12
JournalJournal of the American Chemical Society
Volume135
Issue number42
DOIs
StatePublished - Oct 31 2013

Fingerprint

formic acid
Oxygenases
Peroxides
Aldehydes
Oxygen
Substrates
Nostoc
Alkanes
Alkenes
Ferritins
Oxidation-Reduction
Transition metals
Metals
Paraffins
Olefins
Electrons
Atoms
Oxidation
Enzymes
Water

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

@article{ab2c06813fe84184852528a39de6343c,
title = "Substrate-triggered addition of dioxygen to the diferrous cofactor of aldehyde-deformylating oxygenase to form a diferric-peroxide intermediate",
abstract = "Cyanobacterial aldehyde-deformylating oxygenases (ADOs) belong to the ferritin-like diiron-carboxylate superfamily of dioxygen-activating proteins. They catalyze conversion of saturated or monounsaturated Cn fatty aldehydes to formate and the corresponding Cn-1 alkanes or alkenes, respectively. This unusual, apparently redox-neutral transformation actually requires four electrons per turnover to reduce the O2 cosubstrate to the oxidation state of water and incorporates one O-atom from O2 into the formate coproduct. We show here that the complex of the diiron(II/II) form of ADO from Nostoc punctiforme (Np) with an aldehyde substrate reacts with O2 to form a colored intermediate with spectroscopic properties suggestive of a Fe2III/III complex with a bound peroxide. Its M{\"o}ssbauer spectra reveal that the intermediate possesses an antiferromagnetically (AF) coupled Fe2III/III center with resolved subsites. The intermediate is long-lived in the absence of a reducing system, decaying slowly (t1/2 ∼ 400 s at 5 C) to produce a very modest yield of formate (<0.15 enzyme equivalents), but reacts rapidly with the fully reduced form of 1-methoxy-5-methylphenazinium methylsulfate ( MeOPMS) to yield product, albeit at only ∼50{\%} of the maximum theoretical yield (owing to competition from one or more unproductive pathway). The results represent the most definitive evidence to date that ADO can use a diiron cofactor (rather than a homo- or heterodinuclear cluster involving another transition metal) and provide support for a mechanism involving attack on the carbonyl of the bound substrate by the reduced O2 moiety to form a Fe2III/III-peroxyhemiacetal complex, which undergoes reductive O-O-bond cleavage, leading to C1-C2 radical fragmentation and formation of the alk(a/e)ne and formate products.",
author = "Pandelia, {Maria E.} and Ning Li and Hanne N{\o}rgaard and Douglas Warui and Rajakovich, {Lauren J.} and Chang, {Wei Chen} and Booker, {Squire J.} and Carsten Krebs and {Bollinger, Jr.}, {Joseph M.}",
year = "2013",
month = "10",
day = "31",
doi = "10.1021/ja405047b",
language = "English (US)",
volume = "135",
pages = "15801--15812",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "42",

}

Substrate-triggered addition of dioxygen to the diferrous cofactor of aldehyde-deformylating oxygenase to form a diferric-peroxide intermediate. / Pandelia, Maria E.; Li, Ning; Nørgaard, Hanne; Warui, Douglas; Rajakovich, Lauren J.; Chang, Wei Chen; Booker, Squire J.; Krebs, Carsten; Bollinger, Jr., Joseph M.

In: Journal of the American Chemical Society, Vol. 135, No. 42, 31.10.2013, p. 15801-15812.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Substrate-triggered addition of dioxygen to the diferrous cofactor of aldehyde-deformylating oxygenase to form a diferric-peroxide intermediate

AU - Pandelia, Maria E.

AU - Li, Ning

AU - Nørgaard, Hanne

AU - Warui, Douglas

AU - Rajakovich, Lauren J.

AU - Chang, Wei Chen

AU - Booker, Squire J.

AU - Krebs, Carsten

AU - Bollinger, Jr., Joseph M.

PY - 2013/10/31

Y1 - 2013/10/31

N2 - Cyanobacterial aldehyde-deformylating oxygenases (ADOs) belong to the ferritin-like diiron-carboxylate superfamily of dioxygen-activating proteins. They catalyze conversion of saturated or monounsaturated Cn fatty aldehydes to formate and the corresponding Cn-1 alkanes or alkenes, respectively. This unusual, apparently redox-neutral transformation actually requires four electrons per turnover to reduce the O2 cosubstrate to the oxidation state of water and incorporates one O-atom from O2 into the formate coproduct. We show here that the complex of the diiron(II/II) form of ADO from Nostoc punctiforme (Np) with an aldehyde substrate reacts with O2 to form a colored intermediate with spectroscopic properties suggestive of a Fe2III/III complex with a bound peroxide. Its Mössbauer spectra reveal that the intermediate possesses an antiferromagnetically (AF) coupled Fe2III/III center with resolved subsites. The intermediate is long-lived in the absence of a reducing system, decaying slowly (t1/2 ∼ 400 s at 5 C) to produce a very modest yield of formate (<0.15 enzyme equivalents), but reacts rapidly with the fully reduced form of 1-methoxy-5-methylphenazinium methylsulfate ( MeOPMS) to yield product, albeit at only ∼50% of the maximum theoretical yield (owing to competition from one or more unproductive pathway). The results represent the most definitive evidence to date that ADO can use a diiron cofactor (rather than a homo- or heterodinuclear cluster involving another transition metal) and provide support for a mechanism involving attack on the carbonyl of the bound substrate by the reduced O2 moiety to form a Fe2III/III-peroxyhemiacetal complex, which undergoes reductive O-O-bond cleavage, leading to C1-C2 radical fragmentation and formation of the alk(a/e)ne and formate products.

AB - Cyanobacterial aldehyde-deformylating oxygenases (ADOs) belong to the ferritin-like diiron-carboxylate superfamily of dioxygen-activating proteins. They catalyze conversion of saturated or monounsaturated Cn fatty aldehydes to formate and the corresponding Cn-1 alkanes or alkenes, respectively. This unusual, apparently redox-neutral transformation actually requires four electrons per turnover to reduce the O2 cosubstrate to the oxidation state of water and incorporates one O-atom from O2 into the formate coproduct. We show here that the complex of the diiron(II/II) form of ADO from Nostoc punctiforme (Np) with an aldehyde substrate reacts with O2 to form a colored intermediate with spectroscopic properties suggestive of a Fe2III/III complex with a bound peroxide. Its Mössbauer spectra reveal that the intermediate possesses an antiferromagnetically (AF) coupled Fe2III/III center with resolved subsites. The intermediate is long-lived in the absence of a reducing system, decaying slowly (t1/2 ∼ 400 s at 5 C) to produce a very modest yield of formate (<0.15 enzyme equivalents), but reacts rapidly with the fully reduced form of 1-methoxy-5-methylphenazinium methylsulfate ( MeOPMS) to yield product, albeit at only ∼50% of the maximum theoretical yield (owing to competition from one or more unproductive pathway). The results represent the most definitive evidence to date that ADO can use a diiron cofactor (rather than a homo- or heterodinuclear cluster involving another transition metal) and provide support for a mechanism involving attack on the carbonyl of the bound substrate by the reduced O2 moiety to form a Fe2III/III-peroxyhemiacetal complex, which undergoes reductive O-O-bond cleavage, leading to C1-C2 radical fragmentation and formation of the alk(a/e)ne and formate products.

UR - http://www.scopus.com/inward/record.url?scp=84886506485&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84886506485&partnerID=8YFLogxK

U2 - 10.1021/ja405047b

DO - 10.1021/ja405047b

M3 - Article

C2 - 23987523

AN - SCOPUS:84886506485

VL - 135

SP - 15801

EP - 15812

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 42

ER -