Identification of the Aldo-Keto Reductase Responsible for d-Galacturonic Acid Conversion to l-Galactonate in Saccharomyces cerevisiae

d-galacturonic acid (d-GalUA) is the main constituent of pectin, a complex polysaccharide abundant in several agro-industrial by-products such as sugar beet pulp...

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Autores principales:	Dorthe Rippert, Federica Linguardo, Andreea Perpelea, Mathias Klein, Elke Nevoigt
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<span style="font-variant: small-caps">d</span>-galacturonic acid <span style="font-variant: small-caps">l</span>-galactonate <i>Saccharomyces cerevisiae</i> aldo-keto reductase Gcy1 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/26811f17f9b84ee598ad19aa402b4a6e
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spelling	oai:doaj.org-article:26811f17f9b84ee598ad19aa402b4a6e2021-11-25T18:05:41ZIdentification of the Aldo-Keto Reductase Responsible for <span style="font-variant: small-caps">d</span>-Galacturonic Acid Conversion to <span style="font-variant: small-caps">l</span>-Galactonate in <i>Saccharomyces cerevisiae</i>10.3390/jof71109142309-608Xhttps://doaj.org/article/26811f17f9b84ee598ad19aa402b4a6e2021-10-01T00:00:00Zhttps://www.mdpi.com/2309-608X/7/11/914https://doaj.org/toc/2309-608X<span style="font-variant: small-caps;">d</span>-galacturonic acid (<span style="font-variant: small-caps;">d</span>-GalUA) is the main constituent of pectin, a complex polysaccharide abundant in several agro-industrial by-products such as sugar beet pulp or citrus peel. During several attempts to valorise <span style="font-variant: small-caps;">d</span>-GalUA by engineering the popular cell factory <i>Saccharomyces cerevisiae</i>, it became obvious that <span style="font-variant: small-caps;">d-</span>GalUA is, to a certain degree, converted to <span style="font-variant: small-caps;">l</span>-galactonate<span style="font-variant: small-caps;"> (l</span>-GalA) by an endogenous enzymatic activity. The goal of the current work was to clarify the identity of the responsible enzyme(s). A protein homology search identified three NADPH-dependent unspecific aldo-keto reductases in baker’s yeast (encoded by <i>GCY1</i>, <i>YPR1</i> and <i>GRE3</i>) that show sequence similarities to known <span style="font-variant: small-caps;">d</span>-GalUA reductases from filamentous fungi. Characterization of the respective deletion mutants and an in vitro enzyme assay with a Gcy1 overproducing strain verified that Gcy1 is mainly responsible for the detectable reduction of <span style="font-variant: small-caps;">d</span>-GalUA to <span style="font-variant: small-caps;">l</span>-GalA.Dorthe RippertFederica LinguardoAndreea PerpeleaMathias KleinElke NevoigtMDPI AGarticle<span style="font-variant: small-caps">d</span>-galacturonic acid<span style="font-variant: small-caps">l</span>-galactonate<i>Saccharomyces cerevisiae</i>aldo-keto reductaseGcy1Biology (General)QH301-705.5ENJournal of Fungi, Vol 7, Iss 914, p 914 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<span style="font-variant: small-caps">d</span>-galacturonic acid <span style="font-variant: small-caps">l</span>-galactonate <i>Saccharomyces cerevisiae</i> aldo-keto reductase Gcy1 Biology (General) QH301-705.5
spellingShingle	<span style="font-variant: small-caps">d</span>-galacturonic acid <span style="font-variant: small-caps">l</span>-galactonate <i>Saccharomyces cerevisiae</i> aldo-keto reductase Gcy1 Biology (General) QH301-705.5 Dorthe Rippert Federica Linguardo Andreea Perpelea Mathias Klein Elke Nevoigt Identification of the Aldo-Keto Reductase Responsible for <span style="font-variant: small-caps">d</span>-Galacturonic Acid Conversion to <span style="font-variant: small-caps">l</span>-Galactonate in <i>Saccharomyces cerevisiae</i>
description	<span style="font-variant: small-caps;">d</span>-galacturonic acid (<span style="font-variant: small-caps;">d</span>-GalUA) is the main constituent of pectin, a complex polysaccharide abundant in several agro-industrial by-products such as sugar beet pulp or citrus peel. During several attempts to valorise <span style="font-variant: small-caps;">d</span>-GalUA by engineering the popular cell factory <i>Saccharomyces cerevisiae</i>, it became obvious that <span style="font-variant: small-caps;">d-</span>GalUA is, to a certain degree, converted to <span style="font-variant: small-caps;">l</span>-galactonate<span style="font-variant: small-caps;"> (l</span>-GalA) by an endogenous enzymatic activity. The goal of the current work was to clarify the identity of the responsible enzyme(s). A protein homology search identified three NADPH-dependent unspecific aldo-keto reductases in baker’s yeast (encoded by <i>GCY1</i>, <i>YPR1</i> and <i>GRE3</i>) that show sequence similarities to known <span style="font-variant: small-caps;">d</span>-GalUA reductases from filamentous fungi. Characterization of the respective deletion mutants and an in vitro enzyme assay with a Gcy1 overproducing strain verified that Gcy1 is mainly responsible for the detectable reduction of <span style="font-variant: small-caps;">d</span>-GalUA to <span style="font-variant: small-caps;">l</span>-GalA.
format	article
author	Dorthe Rippert Federica Linguardo Andreea Perpelea Mathias Klein Elke Nevoigt
author_facet	Dorthe Rippert Federica Linguardo Andreea Perpelea Mathias Klein Elke Nevoigt
author_sort	Dorthe Rippert
title	Identification of the Aldo-Keto Reductase Responsible for <span style="font-variant: small-caps">d</span>-Galacturonic Acid Conversion to <span style="font-variant: small-caps">l</span>-Galactonate in <i>Saccharomyces cerevisiae</i>
title_short	Identification of the Aldo-Keto Reductase Responsible for <span style="font-variant: small-caps">d</span>-Galacturonic Acid Conversion to <span style="font-variant: small-caps">l</span>-Galactonate in <i>Saccharomyces cerevisiae</i>
title_full	Identification of the Aldo-Keto Reductase Responsible for <span style="font-variant: small-caps">d</span>-Galacturonic Acid Conversion to <span style="font-variant: small-caps">l</span>-Galactonate in <i>Saccharomyces cerevisiae</i>
title_fullStr	Identification of the Aldo-Keto Reductase Responsible for <span style="font-variant: small-caps">d</span>-Galacturonic Acid Conversion to <span style="font-variant: small-caps">l</span>-Galactonate in <i>Saccharomyces cerevisiae</i>
title_full_unstemmed	Identification of the Aldo-Keto Reductase Responsible for <span style="font-variant: small-caps">d</span>-Galacturonic Acid Conversion to <span style="font-variant: small-caps">l</span>-Galactonate in <i>Saccharomyces cerevisiae</i>
title_sort	identification of the aldo-keto reductase responsible for <span style="font-variant: small-caps">d</span>-galacturonic acid conversion to <span style="font-variant: small-caps">l</span>-galactonate in <i>saccharomyces cerevisiae</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/26811f17f9b84ee598ad19aa402b4a6e
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Identification of the Aldo-Keto Reductase Responsible for <span style="font-variant: small-caps">d</span>-Galacturonic Acid Conversion to <span style="font-variant: small-caps">l</span>-Galactonate in <i>Saccharomyces cerevisiae</i>

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