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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 or citrus peel. During several attempts to valorise  d -GalUA by engineering the popular cell factory  Saccharomyces cerevisiae , it became obvious that  d- GalUA is, to a certain degree, converted to  l -galactonate  (l -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  GCY1 ,  YPR1  and  GRE3 ) that show sequence similarities to known  d -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  d -GalUA to  l -GalA.

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