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Molecular Biology Reports

, Volume 46, Issue 6, pp 6311–6323 | Cite as

Characterization of two Vitis vinifera carotenoid cleavage dioxygenases by heterologous expression in Saccharomyces cerevisiae

  • Nan Meng
  • Guo-Liang Yan
  • Dan Zhang
  • Xiang-Yi Li
  • Chang-Qing Duan
  • Qiu-Hong PanEmail author
Original Article
  • 124 Downloads

Abstract

Norisoprenoids are produced from carotenoids under oxidative degradation mediated by carotenoid cleavage dioxygenases (CCDs) and contribute to floral and fruity notes in grape berries and wine. The diversity of CCD substrates and products has been demonstrated by in vitro recombinant proteins extracted from Escherichia coli expressing CCD genes and of in vivo proteins in an E. coli system co-expressing genes for carotenoid synthesis and cleavage. In the current study, VvCCD1 and VvCCD4b were isolated from the cDNA library of Vitis vinifera L. cv. Cabernet Sauvignon and then transformed into carotenoid-accumulating recombinant Saccharomyces cerevisiae strains. The expression of the target genes was monitored during the yeast growth period, and the accumulation of carotenoids and norisoprenoids in the recombinant strains was measured. The results indicated that both of the VvCCDs cleaved β-carotene at the 7, 8 (7′, 8′) position into β-cyclocitral for the first time. Additionally, the two enzymes also degraded β-carotene at the 9, 10 (9′, 10′) position to generate β-ionone and cleaved lycopene at the 5, 6 (5′, 6′) position into 6-methyl-5-hepten-2-one. These findings suggested that the VvCCDs may possess more cleavage characteristics under the eukaryotic expression system in S. cerevisiae than the prokaryotic system in E. coli, which could better explain the biochemical functions of VvCCDs in grape berries.

Keywords

Carotenoid cleavage dioxygenase Vitis vinifera L cv. Cabernet Sauvignon Carotenoids Norisoprenoids Saccharomyces cerevisiae 

Notes

Author Contributions

Project administration, QHP; Formal analysis and writing - original draft, NM; Writing - review & editing, GLY, CQD and QHP; Methodology, DZ; Data curation, XYL.

Funding

This research was funded by National Natural Science Foundation of China, Grant Number 31272118, and the China Agriculture Research System (CARS-29). Qiu-Hong Pan received the funding. The funders had no role in study design, data collection and analysis, preparation of manuscript, and decision on publication.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11033_2019_5072_MOESM1_ESM.docx (6.3 mb)
Supplementary material 1 (DOCX 6,478 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Nan Meng
    • 1
    • 2
  • Guo-Liang Yan
    • 1
    • 2
  • Dan Zhang
    • 1
    • 2
  • Xiang-Yi Li
    • 1
    • 2
  • Chang-Qing Duan
    • 1
    • 2
  • Qiu-Hong Pan
    • 1
    • 2
    Email author
  1. 1.Center for Viticulture and Enology, College of Food Science & Nutritional EngineeringChina Agricultural UniversityBeijingChina
  2. 2.Key Laboratory of Viticulture and EnologyMinistry of Agricultural and Rural AffairsBeijingChina

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