Combinatorial engineering of betalain biosynthesis pathway in yeast Saccharomyces cerevisiae

Mahsa Babaei, Philip Tinggaard Thomsen, Jane Dannow Dyekjær, Christiane Ursula Glitz, Marc Cernuda Pastor, Peter Gockel, Johann Dietmar Körner, Daniela Rago, Irina Borodina*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Background:
Betalains, comprising red–violet betacyanins and yellow–orange betaxanthins, are the hydrophilic vacuolar pigments that provide bright coloration to roots, fruits, and flowers of plants of the Caryophyllales order. Betanin extracted from red beets is permitted quantum satis as a natural red food colorant (E162). Due to antioxidant activity, betanin has potential health benefits. 

Results:
We applied combinatorial engineering to find the optimal combination of a dozen tyrosine hydroxylase (TyH) and 4,5-dopa-estradiol-dioxygenase (DOD) variants. The best-engineered Saccharomyces cerevisiae strains produced over six-fold higher betaxanthins than previously reported. By genome-resequencing of these strains, we found out that two copies of DOD enzyme from Bougainvillea glabra together with TyH enzymes from Abronia nealleyi, Acleisanthes obtusa, and Cleretum bellidiforme were present in the three high-betaxanthin-producing isolates. Next, we expressed four variants of glucosyltransferases from Beta vulgaris for betanin biosynthesis. The highest titer of betanin (30.8 ± 0.14 mg/L after 48 h from 20 g/L glucose) was obtained when completing the biosynthesis pathway with UGT73A36 glucosyltransferase from Beta vulgaris. Finally, we investigated betalain transport in CEN.PK and S288C strains of Saccharomyces cerevisiae and identified a possible role of transporter genes QDR2 and APL1 in betanin transport.

Conclusions:
This study shows the potential of combinatorial engineering of yeast cell factories for the biotechnological production of betanin.

Original languageEnglish
Article number128
JournalBiotechnology for Biofuels and Bioproducts
Volume16
Issue number1
DOIs
Publication statusPublished - 2023

Keywords

  • Betalains
  • Betanin
  • Betaxanthins
  • Metabolic engineering
  • Saccharomyces cerevisiae

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