TY - JOUR
T1 - Combinatorial engineering of betalain biosynthesis pathway in yeast Saccharomyces cerevisiae
AU - Babaei, Mahsa
AU - Thomsen, Philip Tinggaard
AU - Dyekjær, Jane Dannow
AU - Glitz, Christiane Ursula
AU - Pastor, Marc Cernuda
AU - Gockel, Peter
AU - Körner, Johann Dietmar
AU - Rago, Daniela
AU - Borodina, Irina
N1 - Publisher Copyright:
© 2023, BioMed Central Ltd., part of Springer Nature.
PY - 2023
Y1 - 2023
N2 - 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.
AB - 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.
KW - Betalains
KW - Betanin
KW - Betaxanthins
KW - Metabolic engineering
KW - Saccharomyces cerevisiae
U2 - 10.1186/s13068-023-02374-4
DO - 10.1186/s13068-023-02374-4
M3 - Journal article
C2 - 37592353
AN - SCOPUS:85168411299
VL - 16
JO - Biotechnology for Biofuels and Bioproducts
JF - Biotechnology for Biofuels and Bioproducts
IS - 1
M1 - 128
ER -