Enantiocomplementary Yarrowia lipolytica Oxidoreductases: Alcohol Dehydrogenase 2 and Short Chain Dehydrogenase/Reductase

Abstract

Enzymes of the non-conventional yeast Yarrowia lipolytica seem to be tailor-made for the conversion of lipophilic substrates. Herein, we cloned and overexpressed the Zn-dependent alcohol dehydrogenase ADH2 from Yarrowia lipolytica in Escherichia coli. The purified enzyme was characterized in vitro. The substrate scope for YlADH2 mediated oxidation and reduction was investigated spectrophotometrically and the enzyme showed a broader substrate range than its homolog from Saccharomyces cerevisiae. A preference for secondary compared to primary alcohols in oxidation direction was observed for YlADH2. 2-Octanone was investigated in reduction mode in detail. Remarkably, YlADH2 displays perfect (S)-selectivity and together with a highly (R)-selective short chain dehydrogenase/ reductase from Yarrowia lipolytica it is possible to access both enantiomers of 2-octanol in >99% ee with Yarrowia lipolytica oxidoreductases.

Yeast-Mediated Stereoselective Reduction of α-Acetylbutyrolactone

“…In the Y. lipolytica strains, stereoselective dehydrogenases have been identified [25]. We assumed based on this statement and Ribeiro's work [2], that the reduction of 1 would therefore be catalysed by dehydrogenase.…”

“…1 H NMR (500 MHz, CD 3 OD, Figure S2): 1. 25 The configuration of the individual enantiomers of α'-1'-hydroxyethyl-γ-butyrolactone 2 was determined by comparing it to the biotransformation of 1 by the Yarrowia lipolytica strain found in literature data [18] and results of biotransformation 1 by Rhodotorula marina AM77 described in the Supplementary Materials. (Section S1 and Figure S1)…”

Sequential chemo–biocatalytic synthesis of aroma compounds

“…A similar behaviour regarding the influence of the pH on ADH activity has been reported previously for other ADHs. 61–63…”

Validated Growth Rate-Dependent Regulation of Lipid Metabolism in Yarrowia lipolytica

“…Among the remaining upregulated proteins at maximized lipid yield were several known dehydrogenases, including formate dehydrogenase (FDH) and alcohol dehydrogenase (ADH). Both enzymes are well known for their role in cofactor recovery, t mainly to promote NAD(P)H-dependent enzymes that are in high demand for lipid biosynthesis [ 2 , 11 ]. Moreover, conversion of dihydroxyacetone phosphate (DHAP) to glyceraldehyde-3-phosphate (G3P) by upregulated triose phosphate isomerase could possibly enhance lipid accumulation.…”