Characterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1.

A novel flavoprotein monooxygenase, trans-anethole oxygenase (TAO), from Pseudomonas putida JYR-1, which is capable of catalyzing the oxidation of trans-anethole to p-anisaldehyde, was heterologously expressed in E. coli and purified. Enzymatic kinetics of diverse substrates and cofactors revealed t...

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Autores principales:	Dongfei Han, Michael J Sadowsky, Youhoon Chong, Hor-Gil Hur
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d5ab40e9a7c949f7b70a296e891147df
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spelling	oai:doaj.org-article:d5ab40e9a7c949f7b70a296e891147df2021-11-18T08:55:13ZCharacterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1.1932-620310.1371/journal.pone.0073350https://doaj.org/article/d5ab40e9a7c949f7b70a296e891147df2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24066043/?tool=EBIhttps://doaj.org/toc/1932-6203A novel flavoprotein monooxygenase, trans-anethole oxygenase (TAO), from Pseudomonas putida JYR-1, which is capable of catalyzing the oxidation of trans-anethole to p-anisaldehyde, was heterologously expressed in E. coli and purified. Enzymatic kinetics of diverse substrates and cofactors revealed that TAO is likely to be a novel self-sufficient flavoprotein monooxygenase. Enzyme assays of GST-TAO demonstrated that TAO catalyzed a trans-anethole oxidation reaction without auxiliary component enzyme-like electron-transfer flavin reductases. The single component TAO had the ability to reduce flavin cofactors and simultaneously oxidize trans-anthole to p-anisaldehyde. In the processes of reduction of flavin and oxidation of trans-anethole, TAO accepted various flavin and NAD(P)H cofactors. TAO also catalyzed oxidation of isoeugenol, O-methyl isoeugenol, and isosafrole, all of which contain the 2-propenyl functional group on the aromatic ring structure with different catalytic efficiency. TAO had the greatest catalytic efficiency (k cat/K m) with the original substrate, trans-anethole. Investigation about partially deleted mutants of TAO indicated that reductase active sites appeared to be located near the N terminal. Site directed mutagenesis studies also proved that the proposed flavin binding sites, Trp-38, Thr-43, Tyr-55, were critical for flavin reduction. However, disruption of any portion of TAO eliminated the oxygenase activity.Dongfei HanMichael J SadowskyYouhoon ChongHor-Gil HurPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 9, p e73350 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Dongfei Han Michael J Sadowsky Youhoon Chong Hor-Gil Hur Characterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1.
description	A novel flavoprotein monooxygenase, trans-anethole oxygenase (TAO), from Pseudomonas putida JYR-1, which is capable of catalyzing the oxidation of trans-anethole to p-anisaldehyde, was heterologously expressed in E. coli and purified. Enzymatic kinetics of diverse substrates and cofactors revealed that TAO is likely to be a novel self-sufficient flavoprotein monooxygenase. Enzyme assays of GST-TAO demonstrated that TAO catalyzed a trans-anethole oxidation reaction without auxiliary component enzyme-like electron-transfer flavin reductases. The single component TAO had the ability to reduce flavin cofactors and simultaneously oxidize trans-anthole to p-anisaldehyde. In the processes of reduction of flavin and oxidation of trans-anethole, TAO accepted various flavin and NAD(P)H cofactors. TAO also catalyzed oxidation of isoeugenol, O-methyl isoeugenol, and isosafrole, all of which contain the 2-propenyl functional group on the aromatic ring structure with different catalytic efficiency. TAO had the greatest catalytic efficiency (k cat/K m) with the original substrate, trans-anethole. Investigation about partially deleted mutants of TAO indicated that reductase active sites appeared to be located near the N terminal. Site directed mutagenesis studies also proved that the proposed flavin binding sites, Trp-38, Thr-43, Tyr-55, were critical for flavin reduction. However, disruption of any portion of TAO eliminated the oxygenase activity.
format	article
author	Dongfei Han Michael J Sadowsky Youhoon Chong Hor-Gil Hur
author_facet	Dongfei Han Michael J Sadowsky Youhoon Chong Hor-Gil Hur
author_sort	Dongfei Han
title	Characterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1.
title_short	Characterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1.
title_full	Characterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1.
title_fullStr	Characterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1.
title_full_unstemmed	Characterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1.
title_sort	characterization of a self-sufficient trans-anethole oxygenase from pseudomonas putida jyr-1.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/d5ab40e9a7c949f7b70a296e891147df
work_keys_str_mv	AT dongfeihan characterizationofaselfsufficienttransanetholeoxygenasefrompseudomonasputidajyr1 AT michaeljsadowsky characterizationofaselfsufficienttransanetholeoxygenasefrompseudomonasputidajyr1 AT youhoonchong characterizationofaselfsufficienttransanetholeoxygenasefrompseudomonasputidajyr1 AT horgilhur characterizationofaselfsufficienttransanetholeoxygenasefrompseudomonasputidajyr1
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Characterization of a self-sufficient trans-anethole oxygenase from Pseudomonas putida JYR-1.

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