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>
<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...
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| Autores principales: | , , , , |
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| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
MDPI AG
2021
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| Materias: | |
| Acceso en línea: | https://doaj.org/article/26811f17f9b84ee598ad19aa402b4a6e |
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| Sumario: | <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. |
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