Characterization of Type IV Carboxylate Reductases (CARs) for Whole Cell-Mediated Preparation of 3-Hydroxytyrosol

Melissa Horvat, Susanne Fritsche, Robert Kourist, Margit Winkler

Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

Abstract

Fragrance and flavor industries could not imagine business without aldehydes. Processes for their commercial production raise environmental and ecological concerns. The chemical reduction of organic acids to aldehydes is challenging. To fulfill the demand of a mild and selective reduction of carboxylic acids to aldehydes, carboxylic acid reductases (CARs) are gaining importance. We identified two new subtype IV fungal CARs from Dichomitus squalens CAR (DsCAR) and Trametes versicolor CAR (Tv2CAR) in addition to literature known Trametes versicolor CAR (TvCAR). Expression levels were improved by the co-expression of GroEL-GroES with either the trigger factor or the DnaJ-DnaK-GrpE system. Investigation of the substrate scope of the three enzymes revealed overlapping substrate-specificities. Tv2CAR and DsCAR showed a preferred pH range of 7.0 to 8.0 in bicine buffer. TvCAR showed highest activity at pH 6.5 to 7.5 in MES buffer and slightly reduced activity at pH 6.0 or 8.0. TvCAR appeared to tolerate a wider pH range without significant loss of activity. Type IV fungal CARs optimal temperature was in the range of 25–35 °C. TvCAR showed a melting temperature (T m ) of 55 °C indicating higher stability compared to type III and the other type IV fungal CARs (T m 51–52 °C). Finally, TvCAR was used as the key enzyme for the bioreduction of 3,4-dihydroxyphenylacetic acid to the antioxidant 3-hydroxytyrosol (3-HT) and gave 58 mM of 3-HT after 24 h, which correlates to a productivity of 0.37 g L −1 h −1 .

Originalspracheenglisch
Seiten (von - bis)4171-4181
FachzeitschriftChemCatChem
DOIs
PublikationsstatusElektronische Veröffentlichung vor Drucklegung. - 19 Mär 2019

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carboxylates
Oxidoreductases
preparation
cells
aldehydes
Aldehydes
carboxylic acids
enzymes
Carboxylic acids
Buffers
buffers
Enzymes
Fragrances
3,4-Dihydroxyphenylacetic Acid
acids
Flavors
Organic acids
antioxidants
Substrates
Carboxylic Acids

Schlagwörter

    ASJC Scopus subject areas

    • !!Catalysis
    • Anorganische Chemie
    • !!Physical and Theoretical Chemistry
    • Organische Chemie

    Fields of Expertise

    • Human- & Biotechnology

    Dies zitieren

    Characterization of Type IV Carboxylate Reductases (CARs) for Whole Cell-Mediated Preparation of 3-Hydroxytyrosol. / Horvat, Melissa; Fritsche, Susanne; Kourist, Robert; Winkler, Margit.

    in: ChemCatChem, 19.03.2019, S. 4171-4181.

    Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

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