Production of a recombinant carotenoid cleavage dioxygenase from grape and enzyme assay in water-miscible organic solvents

Mathieu, Sandrine; Bigey, Frédéric; Procureur, Jérôme; Terrier, Nancy; Günata, Ziya

doi:10.1007/s10529-007-9315-8

Cited by 24 publications

(17 citation statements)

References 15 publications

(25 reference statements)

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“…6A, lane 4) as has been observed for other plant CCD1 enzymes (34,44). As shown previously, addition of Triton X-100 to lysed cells prior to centrifugation increased soluble protein recovery (22,44).…”

Section: Ccd1 Enzymes From Arabidopsis and Tomato Generatesupporting

confidence: 81%

The Carotenoid Cleavage Dioxygenase 1 Enzyme Has Broad Substrate Specificity, Cleaving Multiple Carotenoids at Two Different Bond Positions

Vogel

Tan

McCarty

et al. 2008

Journal of Biological Chemistry

243

229

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In many organisms, various enzymes mediate site-specific carotenoid cleavage to generate biologically active apocarotenoids. These carotenoid-derived products include provitamin A, hormones, and flavor and fragrance molecules. In plants, the CCD1 enzyme cleaves carotenoids at 9,10 (9,10) bonds to generate multiple apocarotenoid products. Here we systematically analyzed volatile apocarotenoids generated by maize CCD1 (ZmCCD1) from multiple carotenoid substrates. ZmCCD1 did not cleave geranylgeranyl diphosphate or phytoene but did cleave other linear and cyclic carotenoids, producing volatiles derived from 9,10 (9,10) bond cleavage. Additionally the Arabidopsis, maize, and tomato CCD1 enzymes all cleaved lycopene to generate 6-methyl-5-hepten-2-one. 6-Methyl-5-hepten-2-one, an important flavor volatile in tomato, was produced by cleavage of the 5,6 or 5,6 bond positions of lycopene but not geranylgeranyl diphosphate, -carotene, or phytoene. In vitro, ZmCCD1 cleaved linear and cyclic carotenoids with equal efficiency. Based on the pattern of apocarotenoid volatiles produced, we propose that CCD1 recognizes its cleavage site based on the saturation status between carbons 7 and 8 (7 and 8) and carbons 11 and 12 (11 and 12) as well as the methyl groups on carbons 5, 9, and 13 (5, 9, and 13).Apocarotenoids are terpenoid compounds derived from the oxidative cleavage of carotenoids (1). They are generated when double bonds in a carotenoid are cleaved by molecular oxygen, forming an aldehyde or ketone in each product at the site of cleavage. Carotenoids can be cleaved at any of their conjugated double bonds, resulting in a diverse set of apocarotenoids. This structural diversity is the consequence of the large number of carotenoid precursors (more than 600) and subsequent modifications such as oxidation, reduction, and conjugation. Although apocarotenoid formation can also occur via nonspecific oxidation, biologically active forms with regulatory functions are expected to be generated via site-specific cleavage.Apocarotenoids are widely distributed in nature and serve important biological functions. Examples of biologically active apocarotenoids include retinoids in animals (2), trisporic acid in fungi (3), and abscisic acid in higher plants (4). A variety of other biologically important compounds are believed to be derived by oxidative cleavage of carotenoids. Among these compounds are those associated with mycorrhizal colonization, including mycorradicin (5), blumenin (6), and strigolactone (7).Various enzymes mediate the site-specific carotenoid cleavage needed to generate biologically active apocarotenoids. The founding member of the carotenoid-cleaving enzymes is maize VIVIPAROUS14 (VP14) (8, 9). This 9-cis-epoxycarotenoid dioxygenase (NCED) generates abscisic acid via asymmetrical cleavage at the 11,12 double bond of neoxanthin and/or violaxanthin. NCED genes have been identified in a variety of plant species (9 -13). Using sequence similarity to VP14, the 15,15Ј-dioxygenases responsible for vitamin A biosynthesis w...

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Section: Ccd1 Enzymes From Arabidopsis and Tomato Generatesupporting

confidence: 81%

The Carotenoid Cleavage Dioxygenase 1 Enzyme Has Broad Substrate Specificity, Cleaving Multiple Carotenoids at Two Different Bond Positions

Vogel

Tan

McCarty

et al. 2008

Journal of Biological Chemistry

243

229

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“…None of the transcripts encoding isoprenoid or anthocyanin biosynthetic enzymes were detected as decreasing during any stage of ripening initiation tested. Transcripts encoding VvCCD1, a carotenoid cleavage dioxygenase previously shown to be a key downstream step in norisoprenoid biosynthesis in grapevine (Mathieu et al 2007; AY856353), were relatively abundant throughout ripening initiation and exhibited no significant change (data not shown).…”

Section: Phenotypic Characterization Of Ripening Initiationmentioning

confidence: 95%

Gene expression analyses in individual grape (Vitis vinifera L.) berries during ripening initiation reveal that pigmentation intensity is a valid indicator of developmental staging within the cluster

et al. 2008

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Asynchronous ripening of individual grape berries within clusters can lead to inconsistent organoleptic characteristics for wine making. Ripening initiation in grape berries is non-climacteric and not well understood at the molecular level. Evidence is lacking for a single master switch controlling this process, such as the established role for ethylene in climacteric fruit ripening. We used Affymetrix microarray analyses of 32 individual Vitis vinifera cv. Cabernet Sauvignon berries sampled from two clusters at 50% ripening initiation. By delineating four developmental stages of ripening initiation, we demonstrate that pigmentation is a statistically significant indicator of transcriptional state during ripening initiation. We report on clustered gene expression patterns which were mined for genes annotated with signal transduction functions in order to advance regulatory network modeling of ripening initiation in grape berries. Abscisic acid has previously been demonstrated to be an important signaling component regulating ripening initiation in grapevine. We demonstrate via real-time RT-PCR analyses that up-regulation of a 9-cis-epoxycarotenoid gene family member, VvNCED2, in grape seed and pericarp and a putative ortholog to a reported abscisic acid receptor, VvGCR2, are correlated with ripening initiation. Our results suggest a role for these genes in abscisic acid signaling during ripening initiation.

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“…Tissue homogenates were centrifuged at 20,800 g for 20 min at 4 C. Protein content of the total soluble protein (TSP) was estimated by Bradford assay. 84 CCD enzyme activity was measured spectrophotometrically with lutein (Sigma Aldrich, USA) as substrate, according to Mathieu et al, 85 with minor modifications. Lutein was dissolved in 80% acetone (0.5 g.L ¡1 ) to obtain a final concentration of 35 mM.…”

Section: Estimation Of Carotenoid Cleavage Dioxygenase (Ccd) Activitymentioning

confidence: 99%

Nitric oxide mediates strigolactone signaling in auxin and ethylene-sensitive lateral root formation in sunflower seedlings

Bharti

Bhatla²

2015

Plant Signaling & Behavior

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Strigolactones (SLs) play significant role in shaping root architecture whereby auxin-SL crosstalk has been observed in SL-mediated responses of primary root elongation, lateral root formation and adventitious root (AR) initiation. Whereas GR24 (a synthetic strigolactone) inhibits LR and AR formation, the effect of SL biosynthesis inhibitor (fluridone) is just the opposite (root proliferation). Naphthylphthalamic acid (NPA) leads to LR proliferation but completely inhibits AR development. The diffusive distribution of PIN1 in the provascular cells in the differentiating zone of the roots in response to GR24, fluridone or NPA treatments further indicates the involvement of localized auxin accumulation in LR development responses. Inhibition of LR formation by GR24 treatment coincides with inhibition of ACC synthase activity. Profuse LR development by fluridone and NPA treatments correlates with enhanced [Ca 2C ] cyt in the apical region and differentiating zones of LR, indicating a critical role of [Ca 2C ] in LR development in response to the coordinated action of auxins, ethylene and SLs. Significant enhancement of carotenoid cleavage dioxygenase (CCD) activity (enzyme responsible for SL biosynthesis) in tissue homogenates in presence of cPTIO (NO scavenger) indicates the role of endogenous NO as a negative modulator of CCD activity. Differences in the spatial distribution of NO in the primary and lateral roots further highlight the involvement of NO in SL-modulated root morphogenesis in sunflower seedlings. Present work provides new report on the negative modulation of SL biosynthesis through modulation of CCD activity by endogenous nitric oxide during SL-modulated LR development.

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Production of a recombinant carotenoid cleavage dioxygenase from grape and enzyme assay in water-miscible organic solvents

Cited by 24 publications

References 15 publications

The Carotenoid Cleavage Dioxygenase 1 Enzyme Has Broad Substrate Specificity, Cleaving Multiple Carotenoids at Two Different Bond Positions

The Carotenoid Cleavage Dioxygenase 1 Enzyme Has Broad Substrate Specificity, Cleaving Multiple Carotenoids at Two Different Bond Positions

Gene expression analyses in individual grape (Vitis vinifera L.) berries during ripening initiation reveal that pigmentation intensity is a valid indicator of developmental staging within the cluster

Nitric oxide mediates strigolactone signaling in auxin and ethylene-sensitive lateral root formation in sunflower seedlings

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