Functional characterization of enzymes forming volatile esters from strawberry and banana

M.J. Beekwilder, M. Alvarez-Huerta, E.J. Neef, F.W.A. Verstappen, H.J. Bouwmeester, A. Aharoni

    Research output: Contribution to journalArticleAcademicpeer-review

    217 Citations (Scopus)

    Abstract

    Volatile esters are flavor components of the majority of fruits. The last step in their biosynthesis is catalyzed by alcohol acyltransferases (AATs), which link alcohols to acyl moieties. Full-length cDNAs putatively encoding AATs were isolated from fruit of wild strawberry (Fragaria vesca) and banana (Musa sapientum) and compared to the previously isolated SAAT gene from the cultivated strawberry (Fragaria x ananassa). The potential role of these enzymes in fruit flavor formation was assessed. To this end, recombinant enzymes were produced in Escherichia coli, and their activities were analyzed for a variety of alcohol and acyl-CoA substrates. When the results of these activity assays were compared to a phylogenetic analysis of the various members of the acyltransferase family, it was clear that substrate preference could not be predicted on the basis of sequence similarity. In addition, the substrate preference of recombinant enzymes was not necessarily reflected in the representation of esters in the corresponding fruit volatile profiles. This suggests that the specific profile of a given fruit species is to a significant extent determined by the supply of precursors. To study the in planta activity of an alcohol acyltransferase and to assess the potential for metabolic engineering of ester production, we generated transgenic petunia (Petunia hybrida) plants overexpressing the SAAT gene. While the expression of SAAT and the activity of the corresponding enzyme were readily detected in transgenic plants, the volatile profile was found to be unaltered. Feeding of isoamyl alcohol to explants of transgenic lines resulted in the emission of the corresponding acetyl ester. This confirmed that the availability of alcohol substrates is an important parameter to consider when engineering volatile ester formation in plants
    Original languageEnglish
    Pages (from-to)1865-1878
    JournalPlant Physiology
    Volume135
    Issue number4
    DOIs
    Publication statusPublished - 2004

    Fingerprint

    Fragaria
    Musa
    bananas
    strawberries
    Acyltransferases
    acyltransferases
    Esters
    alcohols
    esters
    Alcohols
    Fruit
    Enzymes
    enzymes
    fruits
    Petunia
    flavor
    genetically modified organisms
    isoamyl alcohol
    Musa paradisiaca
    Fragaria vesca

    Keywords

    • floral scent production
    • clarkia-breweri
    • alcohol acyltransferase
    • molecular-cloning
    • linalool synthase
    • escherichia-coli
    • transferase cdna
    • gene-expression
    • messenger-rnas
    • acetyl-coa

    Cite this

    Beekwilder, M. J., Alvarez-Huerta, M., Neef, E. J., Verstappen, F. W. A., Bouwmeester, H. J., & Aharoni, A. (2004). Functional characterization of enzymes forming volatile esters from strawberry and banana. Plant Physiology, 135(4), 1865-1878. https://doi.org/10.1104/pp.104.042580
    Beekwilder, M.J. ; Alvarez-Huerta, M. ; Neef, E.J. ; Verstappen, F.W.A. ; Bouwmeester, H.J. ; Aharoni, A. / Functional characterization of enzymes forming volatile esters from strawberry and banana. In: Plant Physiology. 2004 ; Vol. 135, No. 4. pp. 1865-1878.
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    abstract = "Volatile esters are flavor components of the majority of fruits. The last step in their biosynthesis is catalyzed by alcohol acyltransferases (AATs), which link alcohols to acyl moieties. Full-length cDNAs putatively encoding AATs were isolated from fruit of wild strawberry (Fragaria vesca) and banana (Musa sapientum) and compared to the previously isolated SAAT gene from the cultivated strawberry (Fragaria x ananassa). The potential role of these enzymes in fruit flavor formation was assessed. To this end, recombinant enzymes were produced in Escherichia coli, and their activities were analyzed for a variety of alcohol and acyl-CoA substrates. When the results of these activity assays were compared to a phylogenetic analysis of the various members of the acyltransferase family, it was clear that substrate preference could not be predicted on the basis of sequence similarity. In addition, the substrate preference of recombinant enzymes was not necessarily reflected in the representation of esters in the corresponding fruit volatile profiles. This suggests that the specific profile of a given fruit species is to a significant extent determined by the supply of precursors. To study the in planta activity of an alcohol acyltransferase and to assess the potential for metabolic engineering of ester production, we generated transgenic petunia (Petunia hybrida) plants overexpressing the SAAT gene. While the expression of SAAT and the activity of the corresponding enzyme were readily detected in transgenic plants, the volatile profile was found to be unaltered. Feeding of isoamyl alcohol to explants of transgenic lines resulted in the emission of the corresponding acetyl ester. This confirmed that the availability of alcohol substrates is an important parameter to consider when engineering volatile ester formation in plants",
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    Beekwilder, MJ, Alvarez-Huerta, M, Neef, EJ, Verstappen, FWA, Bouwmeester, HJ & Aharoni, A 2004, 'Functional characterization of enzymes forming volatile esters from strawberry and banana' Plant Physiology, vol. 135, no. 4, pp. 1865-1878. https://doi.org/10.1104/pp.104.042580

    Functional characterization of enzymes forming volatile esters from strawberry and banana. / Beekwilder, M.J.; Alvarez-Huerta, M.; Neef, E.J.; Verstappen, F.W.A.; Bouwmeester, H.J.; Aharoni, A.

    In: Plant Physiology, Vol. 135, No. 4, 2004, p. 1865-1878.

    Research output: Contribution to journalArticleAcademicpeer-review

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    T1 - Functional characterization of enzymes forming volatile esters from strawberry and banana

    AU - Beekwilder, M.J.

    AU - Alvarez-Huerta, M.

    AU - Neef, E.J.

    AU - Verstappen, F.W.A.

    AU - Bouwmeester, H.J.

    AU - Aharoni, A.

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    KW - floral scent production

    KW - clarkia-breweri

    KW - alcohol acyltransferase

    KW - molecular-cloning

    KW - linalool synthase

    KW - escherichia-coli

    KW - transferase cdna

    KW - gene-expression

    KW - messenger-rnas

    KW - acetyl-coa

    U2 - 10.1104/pp.104.042580

    DO - 10.1104/pp.104.042580

    M3 - Article

    VL - 135

    SP - 1865

    EP - 1878

    JO - Plant Physiology

    JF - Plant Physiology

    SN - 0032-0889

    IS - 4

    ER -