Biochemical characterization of the minimal domains of an iterative eukaryotic polyketide synthase

Iterative type I polyketide synthases (PKS) are megaenzymes essential to the biosynthesis of an enormously diverse array of bioactive natural products. Each PKS contains minimally three functional domains, β‐ketosynthase (KS), acyltransferase (AT), and acyl carrier protein (ACP), and a subset of red...

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Detalles Bibliográficos
Autores principales: Sabatini, Martín, Comba, Santiago, Altabe, Silvia Graciela, Recio Balsells, Alejandro Iván, Labadie, Guillermo Roberto, Takano, Eriko, Gramajo, Hugo Cesar, Arabolaza, Ana Lorena
Formato: article artículo publishedVersion
Lenguaje:Inglés
Publicado: Wiley 2020
Materias:
Domain deconstruction
Iterative PKS
PKS biochemistry
Substrate specificity
Acceso en línea:http://hdl.handle.net/2133/18276
http://hdl.handle.net/2133/18276
Aporte de:
Repositorio Hipermedial de la Universidad Nacional de Rosario (UNR) de Universidad Nacional de Rosario
id I15-R121-2133-18276
record_format dspace
institution Universidad Nacional de Rosario
institution_str I-15
repository_str R-121
collection Repositorio Hipermedial de la Universidad Nacional de Rosario (UNR)
language Inglés
orig_language_str_mv eng
topic Domain deconstruction
Iterative PKS
PKS biochemistry
Substrate specificity
spellingShingle Domain deconstruction
Iterative PKS
PKS biochemistry
Substrate specificity
Sabatini, Martín
Comba, Santiago
Altabe, Silvia Graciela
Recio Balsells, Alejandro Iván
Labadie, Guillermo Roberto
Takano, Eriko
Gramajo, Hugo Cesar
Arabolaza, Ana Lorena
Biochemical characterization of the minimal domains of an iterative eukaryotic polyketide synthase
topic_facet Domain deconstruction
Iterative PKS
PKS biochemistry
Substrate specificity
description Iterative type I polyketide synthases (PKS) are megaenzymes essential to the biosynthesis of an enormously diverse array of bioactive natural products. Each PKS contains minimally three functional domains, β‐ketosynthase (KS), acyltransferase (AT), and acyl carrier protein (ACP), and a subset of reducing domains such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER). The substrate selection, condensation reactions, and β‐keto processing of the polyketide growing chain are highly controlled in a programmed manner. However, the structural features and mechanistic rules that orchestrate the iterative cycles, processing domains functionality, and chain termination in this kind of megaenzymes are often poorly understood. Here, we present a biochemical and functional characterization of the KS and the AT domains of a PKS from the mallard duck Anas platyrhynchos (ApPKS). ApPKS belongs to an animal PKS family phylogenetically more related to bacterial PKS than to metazoan fatty acid synthases. Through the dissection of the ApPKS enzyme into mono‐ to didomain fragments and its reconstitution in vitro, we determined its substrate specificity toward different starters and extender units. ApPKS AT domain can effectively transfer acetyl‐CoA and malonyl‐CoA to the ApPKS ACP stand‐alone domain. Furthermore, the KS and KR domains, in the presence of Escherichia coli ACP, acetyl‐CoA, and malonyl‐CoA, showed the ability to catalyze the chain elongation and the β‐keto reduction steps necessary to yield a 3‐hydroxybutyryl‐ACP derivate. These results provide new insights into the catalytic efficiency and specificity of this uncharacterized family of PKSs.
format article
artículo
publishedVersion
author Sabatini, Martín
Comba, Santiago
Altabe, Silvia Graciela
Recio Balsells, Alejandro Iván
Labadie, Guillermo Roberto
Takano, Eriko
Gramajo, Hugo Cesar
Arabolaza, Ana Lorena
author_facet Sabatini, Martín
Comba, Santiago
Altabe, Silvia Graciela
Recio Balsells, Alejandro Iván
Labadie, Guillermo Roberto
Takano, Eriko
Gramajo, Hugo Cesar
Arabolaza, Ana Lorena
author_sort Sabatini, Martín
title Biochemical characterization of the minimal domains of an iterative eukaryotic polyketide synthase
title_short Biochemical characterization of the minimal domains of an iterative eukaryotic polyketide synthase
title_full Biochemical characterization of the minimal domains of an iterative eukaryotic polyketide synthase
title_fullStr Biochemical characterization of the minimal domains of an iterative eukaryotic polyketide synthase
title_full_unstemmed Biochemical characterization of the minimal domains of an iterative eukaryotic polyketide synthase
title_sort biochemical characterization of the minimal domains of an iterative eukaryotic polyketide synthase
publisher Wiley
publishDate 2020
url http://hdl.handle.net/2133/18276
http://hdl.handle.net/2133/18276
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