TY - JOUR

T1 - A DyP-type peroxidase of pleurotus sapidus with alkene cleaving activity

AU - Krahe, Nina Katharina

AU - Berger, Ralf G.

AU - Ersoy, Franziska

N1 - Funding information: This research was funded the BMBF cluster Bioeconomy International 2015, grant number 031B0307A. The APC was funded by the Open Access fund of the Gottfried Wilhelm Leibniz Universität Hannover.

PY - 2020/3/27

Y1 - 2020/3/27

N2 - Alkene cleavage is a possibility to generate aldehydes with olfactory properties for the fragrance and flavor industry. A dye-decolorizing peroxidase (DyP) of the basidiomycete Pleurotus sapidus (PsaPOX) cleaved the aryl alkene trans-anethole. The PsaPOX was semi-purified from the mycelium via FPLC, and the corresponding gene was identified. The amino acid sequence as well as the predicted tertiary structure showed typical characteristics of DyPs as well as a non-canonical Mn 2+-oxidation site on its surface. The gene was expressed in Komagataella pfaffii GS115 yielding activities up to 142 U/L using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) as substrate. PsaPOX exhibited optima at pH 3.5 and 40 °C and showed highest peroxidase activity in the presence of 100 µM H 2O 2 and 25 mM Mn 2+. PsaPOX lacked the typical activity of DyPs towards anthraquinone dyes, but oxidized Mn 2+ to Mn 3+. In addition, bleaching of β-carotene and annatto was observed. Biotransformation experiments verified the alkene cleavage activity towards the aryl alkenes ( E)-methyl isoeugenol, α-methylstyrene, and trans-anethole, which was increased almost twofold in the presence of Mn 2+. The resultant aldehydes are olfactants used in the fragrance and flavor industry. PsaPOX is the first described DyP with alkene cleavage activity towards aryl alkenes and showed potential as biocatalyst for flavor production.

AB - Alkene cleavage is a possibility to generate aldehydes with olfactory properties for the fragrance and flavor industry. A dye-decolorizing peroxidase (DyP) of the basidiomycete Pleurotus sapidus (PsaPOX) cleaved the aryl alkene trans-anethole. The PsaPOX was semi-purified from the mycelium via FPLC, and the corresponding gene was identified. The amino acid sequence as well as the predicted tertiary structure showed typical characteristics of DyPs as well as a non-canonical Mn 2+-oxidation site on its surface. The gene was expressed in Komagataella pfaffii GS115 yielding activities up to 142 U/L using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) as substrate. PsaPOX exhibited optima at pH 3.5 and 40 °C and showed highest peroxidase activity in the presence of 100 µM H 2O 2 and 25 mM Mn 2+. PsaPOX lacked the typical activity of DyPs towards anthraquinone dyes, but oxidized Mn 2+ to Mn 3+. In addition, bleaching of β-carotene and annatto was observed. Biotransformation experiments verified the alkene cleavage activity towards the aryl alkenes ( E)-methyl isoeugenol, α-methylstyrene, and trans-anethole, which was increased almost twofold in the presence of Mn 2+. The resultant aldehydes are olfactants used in the fragrance and flavor industry. PsaPOX is the first described DyP with alkene cleavage activity towards aryl alkenes and showed potential as biocatalyst for flavor production.

KW - Alkene cleavage

KW - Aryl alkenes

KW - Basidiomycota

KW - Biocatalysis

KW - Carotene degradation

KW - Dye-decolorizing peroxidase (DyP)

KW - Komagataella pfaffii

KW - Manganese

KW - Pleurotus sapidus

UR - http://www.scopus.com/inward/record.url?scp=85082731098&partnerID=8YFLogxK

U2 - 10.3390/molecules25071536

DO - 10.3390/molecules25071536

M3 - Article

C2 - 32230972

AN - SCOPUS:85082731098

VL - 25

JO - MOLECULES

JF - MOLECULES

SN - 1420-3049

IS - 7

M1 - 1536

ER -