Biosynthesis of the antioxidant γ-glutamyl-cysteine with engineered Yarrowia lipolytica

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Do D., Guruk M., Kus-Liśkiewicz M., Damblon C., Arguelles-Arias A., Erten H., ...More

BIOTECHNOLOGY JOURNAL, vol.19, no.2300564, pp.1-9, 2024 (SCI-Expanded)

  • Publication Type: Article / Article
  • Volume: 19 Issue: 2300564
  • Publication Date: 2024
  • Doi Number: 10.1002/biot.202300564
  • Journal Name: BIOTECHNOLOGY JOURNAL
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Compendex, EMBASE, MEDLINE, Veterinary Science Database
  • Page Numbers: pp.1-9
  • Çukurova University Affiliated: Yes

Abstract

The dipeptide γ-glutamylcysteine (γ-GC), the first intermediate of glutathione (GSH)

synthesis, is considered as a promising drug to reduce or prevent plethora of agerelated

disorders such as Alzheimer and Parkinson diseases. The unusual γ-linkage

between the two constitutive amino acids, namely cysteine and glutamate, renders

its chemical synthesis particularly challenging. Herein, we report on the metabolic

engineering of the non-conventional yeast Yarrowia lipolytica for efficient γ-GC synthesis.

The yeast was first converted into a γ-GC producer by disruption of gene GSH2

encoding GSH synthase and by constitutive expression of GSH1 encoding glutamylcysteine

ligase. Subsequently genes involved in cysteine and glutamate anabolism,

namely MET4, CYSE, CYSF, and GDH1 were overexpressed with the aim to increase

their intracellular availability. With such a strategy, a γ-GC titer of 464 nmol mg−1

protein (93mg gDCW−1) was obtained within 24 h of cell growth.