Comparative evaluation of secreted plant carotenoid cleavage dioxygenase 1 (CCD1) enzymes in Saccharomyces cerevisiae

John J. B. Timmins*, Heinrich Kroukamp, Roy S. K. Walker, Isak S. Pretorius, Ian T. Paulsen

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)
59 Downloads (Pure)

Abstract

Enabling technologies in synthetic biology now present the opportunity to engineer wine yeast for enhanced novel aromas. In doing so, improved wine products will increase the desirability of wine for the consumer and add value to the winemaker. The action of the enzyme carotenoid cleavage dioxygenase 1 (CCD1) on β-carotene to produce β-ionone is of interest to improve the aroma and flavour of the wine. Engineering the yeast, Saccharomyces cerevisiae, to produce higher concentrations of CCD1 in grape-must presents an opportunity to increase the levels of this volatile organic compound, thus enhancing the organoleptic properties of wine. To this end, four phylogenetically diverse plant CCD1 genes were synthesised with a secretion signal peptide and transformed into S. cerevisiae. The relative ability of each enzyme secreted into the yeast supernatant to cleave the deep orange C40 β-carotene was determined by spectrophotometry; furthermore, the by-product of such cleavage, the highly aromatic C13 β-ionone, was assessed by head-space solid-phase micro-extraction, with analysis and detection by GCMS. Reduction in β-carotene levels and release of β-ionone from the supernatant were validated by LCMS detection of CCD1. These experiments demonstrated that expression in yeast of the CCD1s derived from Petunia hybrida and Vitis vinifera and their subsequent secretion into the medium provided superior efficacy in both β-carotene reduction and β-ionone liberation. We anticipate this knowledge being of benefit to future winemakers in producing a vinous product with enhanced organoleptic properties.

Original languageEnglish
Article number395
Pages (from-to)1-10
Number of pages10
JournalFermentation
Volume8
Issue number8
DOIs
Publication statusPublished - Aug 2022

Bibliographical note

Copyright the Author(s) 2022. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • aroma compounds
  • carotenoid cleavage dioxygenase 1 (CCD1)
  • Saccharomyces cerevisiae
  • synthetic biology
  • wine products
  • β-carotene
  • β-ionone

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  • COESB: ARC Centre of Excellence in Synthetic Biology

    Paulsen, I., Filipovska, A., Parker, R., Nielsen, L. K., Neilan, B. A., Alexandrov, K., Jackson , C., Wodak, J., Rackham, O., Marcellin, E., Gillings, M., Rogers, W., Lee, L., Packer, N., O'Hara, I. M., Speight, R., Vickers, C. E., Beliaev, A., Scott, C., Lacey, J., Mankad, A., Calvert, J., Thomas, G., Rodriguez-Concepcion, M., Fleishman, S., Koepke, M., Ball, M., Turner, N. J., Borneman, A. R., Holowko, M., Goold, H., Ellis, T., Mitchell, L. A. & Dai, J.

    9/11/208/11/27

    Project: Research

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