Secretion of Pseudomonas Aeruginosa Lipoxygenase by Pichia Pastoris upon Glycerol Feed

Chiam Hashem, Holly Stolterfoht, Claudia Rinnofner, Stefan Steinberger, Margit Winkler, Harald Pichler*

*Korrespondierende/r Autor/in für diese Arbeit

Publikation: Beitrag in einer FachzeitschriftArtikel

Abstract

Pseudomonas aeruginosa lipoxygenase (PaLOX) catalyzes the peroxidation of unsaturated fatty acids. Not only linoleic acid, but also linolenic acid and oleic acid are oxidized. The natural host secretes PaLOX into the periplasmic space. Herein, the aim is to secrete PaLOX to the culture supernatant of Pichia pastoris. Since protein background in the culture supernatant is typically rather low, this strategy allows for almost pure production of PaLOX applicable for the valorization of renewable fatty acids, for example for the production of green leaf volatiles. Using the CAT1 promoter system and the well-established α-factor signal sequence for secretion, methanol- and glycerol-induced secretion are compared and the latter shows four times more LOX activity in the culture supernatant under methanol-free conditions. In addition, secreted PaLOX is purified and the specific activity with enzyme in culture supernatant is compared. Notably, the predominant specific activity is achieved for enzyme in culture supernatant - 11.6 U mg −1 - reaching five times higher specific activity than purified PaLOX.

Originalspracheenglisch
Aufsatznummer2000089
FachzeitschriftBiotechnology Journal
Jahrgang15
Ausgabenummer11
Frühes Online-Datum4 Aug 2020
DOIs
PublikationsstatusVeröffentlicht - Nov 2020

ASJC Scopus subject areas

  • !!Applied Microbiology and Biotechnology
  • !!Molecular Medicine

Fields of Expertise

  • Human- & Biotechnology

Kooperationen

  • NAWI Graz
  • BioTechMed-Graz

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