The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Earl H. Harrison - One of the best experts on this subject based on the ideXlab platform.
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limited appearance of Apocarotenoids is observed in plasma after consumption of tomato juices a randomized human clinical trial
The American Journal of Clinical Nutrition, 2018Co-Authors: Jessica L Cooperstone, Janet A Novotny, Ken M Riedl, Morgan J Cichon, David M Francis, Robert W Curley, Steven J Schwartz, Earl H. HarrisonAbstract:Background Nonvitamin A Apocarotenoids occur in foods. Some function as retinoic acid receptor antagonists in vitro, though it is unclear if Apocarotenoids are absorbed or accumulate to levels needed to elicit biological function. Objective The aim of this study was to quantify carotenoids and Apocarotenoids (β-apo-8'-, -10'-, -12'-, and -14'-carotenal, apo-6'-, -8'-, -10'-, -12'-, and -14'-lycopenal, retinal, acycloretinal, β-apo-13-carotenone, and apo-13-lycopenone) in human plasma after controlled consumption of carotenoid-rich tomato juices. Design Healthy subjects (n = 35) consumed a low-carotenoid diet for 2 wk, then consumed 360 mL of high-β-carotene tomato juice (30.4 mg of β-carotene, 34.5 μg total β-Apocarotenoids/d), high-lycopene tomato juice (42.5 mg of lycopene, 119.2 μg total apolycopenoids/d), or a carotenoid-free control (cucumber juice) per day for 4 wk. Plasma was sampled at baseline (after washout) and after 2 and 4 wk, and analyzed for carotenoids and Apocarotenoids using high-pressure liquid chromatography (HPLC) and HPLC-tandem mass spectrometry, respectively. The methods used to analyze the Apocarotenoids had limits of detection of ∼ 100 pmol/L. Results Apocarotenoids are present in tomato juices at 0.1-0.5% of the parent carotenoids. Plasma lycopene and β-carotene increased (P < 0.001) after consuming high-lycopene and β-carotene tomato juices, respectively, while retinol remained unchanged. β-Apo-13-carotenone was found in the blood of all subjects at every visit, although elevated (P < 0.001) after consuming β-carotene tomato juice for 4 wk (1.01 ± 0.27 nmol/L) compared with both baseline (0.37 ± 0.17 nmol/L) and control (0.46 ± 0.11 nmol/L). Apo-6'-lycopenal was detected or quantifiable in 29 subjects, while β-apo-10'- and 12'-carotenal were detected in 6 and 2 subjects, respectively. No other apolycopenoids or Apocarotenoids were detected. Conclusions β-Apo-13-carotenone was the only apocarotenoid that was quantifiable in all subjects, and was elevated in those consuming high-β-carotene tomato juice. Levels were similar to previous reports of all-trans-retinoic acid. Other Apocarotenoids are either poorly absorbed or rapidly metabolized or cleared, and so are absent or limited in blood. β-Apo-13-carotenone may form from vitamin A and its presence warrants further investigation. This trial was registered at clinicaltrials.gov as NCT02550483.
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Apocarotenoids emerging roles in mammals
Annual Review of Nutrition, 2018Co-Authors: Earl H. Harrison, Loredana QuadroAbstract:Apocarotenoids are cleavage products of C40 isoprenoid pigments, named carotenoids, synthesized exclusively by plants and microorganisms. The colors of flowers and fruits and the photosynthetic process are examples of the biological properties conferred by carotenoids to these organisms. Mammals do not synthesize carotenoids but obtain them from foods of plant origin. Apocarotenoids are generated upon enzymatic and nonenzymatic cleavage of the parent compounds both in plants and in the tissues of mammals that have ingested carotenoid-containing foods. The best-characterized Apocarotenoids are retinoids (vitamin A and its derivatives), generated upon central oxidative cleavage of provitamin A carotenoids, mainly β-carotene. In addition to the well-known biological actions of vitamin A, it is becoming apparent that nonretinoid Apocarotenoids also have the potential to regulate a broad spectrum of critical cellular functions, thus influencing mammalian health. This review discusses the current knowledge about the generation and biological activities of nonretinoid Apocarotenoids in mammals.
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Carotenoid metabolism in mammals, including man: formation, occurrence, and function of Apocarotenoids Thematic Review Series: Fat-Soluble Vitamins: Vitamin A
Journal of Lipid Research, 2013Co-Authors: Abdulkerim Eroglu, Earl H. HarrisonAbstract:Vitamin A was recognized as an essential nutrient 100 years ago. In the 1930s, it became clear that dietary β-carotene was cleaved at its central double to yield vitamin A (retinal or β-apo-15'-carotenal). Thus a great deal of research has focused on the central cleavage of provitamin A carotenoids to form vitamin A (retinoids). The mechanisms of formation and the physiological role(s) of noncentral (eccentric) cleavage of both provitamin A carotenoids and nonprovitamin A carotenoids has been less clear. It is becoming apparent that the Apocarotenoids exert unique biological activities themselves. These compounds are found in the diet and thus may be absorbed in the intestine, or they may form from enzymatic or nonenzymatic cleavage of the parent carotenoids. The mechanism of action of Apocarotenoids in mammals is not fully worked out. However, as detailed in this review, they have profound effects on gene expression and work, at least in part, through the modulation of ligand-activated nuclear receptors. Understanding the interactions of Apocarotenoids with other lipid-binding proteins, chaperones, and metabolizing enzymes will undoubtedly increase our understanding of the biological roles of these carotenoid metabolites.
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Thematic Review Series: Fat-Soluble Vitamins: Vitamin A Carotenoid metabolism in mammals, including man: formation, occurrence, and function of Apocarotenoids
2013Co-Authors: Abdulkerim Eroglu, Earl H. HarrisonAbstract:Vitamin A was recognized as an essential nutrient 100 years ago. In the 1930s, it became clear that dietary - carotene was cleaved at its central double to yield vitamin A (retinal or -apo-15 ' -carotenal). Thus a great deal of research has focused on the central cleavage of provitamin A carote- noids to form vitamin A (retinoids). The mechanisms of for- mation and the physiological role(s) of noncentral (eccentric) cleavage of both provitamin A carotenoids and nonprovitamin A carotenoids has been less clear. It is becoming apparent that the Apocarotenoids exert unique biological activities them- selves. These compounds are found in the diet and thus may be absorbed in the intestine, or they may form from enzymatic or nonenzymatic cleavage of the parent carotenoids. The mechanism of action of Apocarotenoids in mammals is not fully worked out. However, as detailed in this review, they have profound effects on gene expression and work, at least in part, through the modulation of ligand-activated nuclear recep- tors. Understanding the interactions of Apocarotenoids with other lipid-binding proteins, chaperones, and metabolizing en- zymes will undoubtedly increase our understanding of the bio- logical roles of these carotenoid metabolites. —Eroglu, A., and E. H. Harrison. Carotenoid metabolism in mammals, including man: formation, occurrence, and function of Apocarotenoids. J. Lipid Res. 2013. 54: 1719-1730.
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The formation, occurrence, and function of β-Apocarotenoids: β-carotene metabolites that may modulate nuclear receptor signaling
The American Journal of Clinical Nutrition, 2012Co-Authors: Earl H. Harrison, Carlo Dela Seña, Abdulkerim Eroglu, Matthew K. FleshmanAbstract:β-Carotene is the major dietary source of provitamin A. Central cleavage of β-carotene yields 2 molecules of retinal followed by further oxidation to retinoic acid. Eccentric cleavage of β-carotene occurs at double bonds other than the central double bond, and the products of these reactions are β-apocarotenals and β-apocarotenones. We reviewed recent developments in 3 areas: 1): the enzymatic production of β-Apocarotenoids in higher animals; 2) the occurrence of β-Apocarotenoids in foods and animal tissues; and 3) the biological activity of β-Apocarotenoids, particularly on retinoid receptors. HPLC–mass spectrometry techniques were developed to quantify these compounds in mouse serum and tissues and in foods. β-Apo-10′- and -12′-carotenals were detected in mouse serum and liver. β-Apo-8′-, β-apo-10′-, β-apo-12′-, and β-apo-14′-carotenals and β-apo-13-carotenone were detected in orange-fleshed melons. Transactivation assays were performed to see whether Apocarotenoids activate or antagonize retinoid X receptor (RXR) α. Reporter gene constructs and retinoid receptor (RXRα) were transfected into cells, which were used to perform quantitative assays for the activation of this ligand-dependent transcription factor. None of the β-Apocarotenoids significantly activated RXRα. However, β-apo-13-carotenone antagonized the 9-cis-retinoic acid activation of RXRα. Competitive radioligand binding assays showed that this antagonist competes directly with the agonist for binding to purified receptor, a finding confirmed by molecular modeling studies. These findings suggest that a possible biological function of β-Apocarotenoids is their ability to interfere with nuclear receptor signaling. Recent work showed that β-apo-13-carotenone is also a high-affinity antagonist of all 3 retinoic acid receptors (RARα, RARβ, and RARγ).
Luigi Mondello - One of the best experts on this subject based on the ideXlab platform.
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Determination of free Apocarotenoids and apocarotenoid esters in human colostrum.
Analytical and Bioanalytical Chemistry, 2020Co-Authors: Mariosimone Zoccali, Daniele Giuffrida, Roberta Granese, Fabio Salafia, Paola Dugo, Luigi MondelloAbstract:The presence of carotenoids in human colostrum has been reported in the literature, and xanthophyll esters in human colostrum were recently detected for the first time. However, no published studies have reported whether Apocarotenoids, which are metabolites derived from carotenoid enzymatic or nonenzymatic oxidative cleavage, are present in human colostrum. Therefore, the purpose of the present study was to search for the possible occurrence of Apocarotenoids, including apocarotenoid esters, in human colostrum for the first time by applying an online supercritical fluid extraction–supercritical fluid chromatography–tandem mass spectrometry methodology. Recent evidence related to apocarotenoid transcriptional activity has suggested that they may have beneficial health properties superior to those of their parent carotenoids. Three different Apocarotenoids, namely apo-8′-β-carotenal, apo-8′-lycopenal, and β-citraurin, were identified in intact human colostrum samples, with average concentrations of 85 nmol L−1, 54.6 nmol L−1, and 75.4 nmol L−1, respectively. The overall detection of 16 different free Apocarotenoids and 10 different apocarotenoid fatty acid esters in human colostrum was achieved here for the first time. Their occurrence in human colostrum certainly has implications for newborn health status, since colostrum is the only form of food for the newborn during the very first days of life.
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First Apocarotenoids Profiling of Four Microalgae Strains
Antioxidants, 2019Co-Authors: Mariosimone Zoccali, Daniele Giuffrida, Fabio Salafia, Paola Dugo, Carmen Socaciu, Kari Skjånes, Luigi MondelloAbstract:Both enzymatic or oxidative carotenoids cleavages can often occur in nature and produce a wide range of bioactive Apocarotenoids. Considering that no detailed information is available in the literature regarding the occurrence of Apocarotenoids in microalgae species, the aim of this study was to study the extraction and characterization of Apocarotenoids in four different microalgae strains: Chlamydomonas sp. CCMP 2294, Tetraselmis chuii SAG 8-6, Nannochloropsis gaditana CCMP 526, and Chlorella sorokiniana NIVA-CHL 176. This was done for the first time using an online method coupling supercritical fluid extraction and supercritical fluid chromatography tandem mass spectrometry. A total of 29 different Apocarotenoids, including various apocarotenoid fatty acid esters, were detected: apo-12’-zeaxanthinal, β-apo-12’-carotenal, apo-12-luteinal, and apo-12’-violaxanthal. These were detected in all the investigated strains together with the two apocarotenoid esters, apo-10’-zeaxanthinal-C4:0 and apo-8’-zeaxanthinal-C8:0. The overall extraction and detection time for the Apocarotenoids was less than 10 min, including Apocarotenoids esters, with an overall analysis time of less than 20 min. Moreover, preliminary quantitative data showed that the β-apo-8’-carotenal content was around 0.8% and 2.4% of the parent carotenoid, in the C. sorokiniana and T. chuii strains, respectively. This methodology could be applied as a selective and efficient method for the Apocarotenoids detection.
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Carotenoids and Apocarotenoids determination in intact human blood samples by online supercritical fluid extraction-supercritical fluid chromatography-tandem mass spectrometry.
Analytica Chimica Acta, 2018Co-Authors: Mariosimone Zoccali, Daniele Giuffrida, Fabio Salafia, Salvatore V Giofre, Luigi MondelloAbstract:Abstract A direct on-line method based on the coupling of supercritical fluid extraction and supercritical fluid chromatography with triple quadrupole mass spectrometry detection (SFE-SFC-QqQ/MS) for selected carotenoids determination and Apocarotenoids detection in intact human blood was developed for the first time. Carotenoids and Apocarotenoids were identified by using the available standard together with full scan, selected ion monitoring (SIM), and multiple reaction monitoring (MRM) experiments. Moreover, β-Cryptoxanthin, Zeaxanthin, β-Carotene and Capsanthin were directly quantified by the developed methodology, using a multiple reaction monitoring (MRM) approach; the determined average content of β-carotene was 123.8 nmol L−1 (range 18.7–485.1 nmol L−1), of β-cryptoxanthin was 385.3 nmol L−1 (range 72.5–1920.3 nmol L−1), of zeaxanthin was 396.9 nmol L−1 (range
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Apocarotenoids determination in capsicum chinense jacq cv habanero by supercritical fluid chromatography triple quadrupole mass spectrometry
Food Chemistry, 2017Co-Authors: Daniele Giuffrida, Mariosimone Zoccali, Paola Dugo, Salvatore V Giofre, Luigi MondelloAbstract:Relatively few studies are available in the literature on the Apocarotenoids occurrence in food which are based on liquid chromatographic separation approaches and no reports are available on the Apocarotenoids separation by SFC-APCI (+/-)/QqQ/MS. The dual purpose of this research was to investigate the Apocarotenoids presence in red habanero peppers and to develop a novel SFC-APCI(+/-)/QqQ/MS methodology, using a novel fused-core C30 column, for the Apocarotenoids detection in a fast and efficient way, for the first time. In this study, 25 different Apocarotenoids were identified, 14 were free Apocarotenoids and 11 were Apocarotenoids fatty acids esters. The methodology allowed for all the separations to occur in less then five minutes. The detected Apo-10'-, Apo-14'- and Apo-15-capsorubinals and different Apo-8'-capsorubinal and Apo-10'-zeaxanthinals fatty acid esters had not been previously tentatively identified in any Capsicum species and, to the best of our knowledge, in any food matrix.
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Apocarotenoids profiling in different Capsicum species.
Food Chemistry, 1Co-Authors: Mariosimone Zoccali, Daniele Giuffrida, Fabio Salafia, Paola Dugo, Francesca Rigano, Monica Casale, Luigi MondelloAbstract:Abstract The present study report on the application of an on line supercritical fluid extraction-supercritical fluid chromatography-triple quadrupole/mass spectrometry methodology to the first Apocarotenoids profiling of seventeen different chilli peppers cultivars belonging to Capsicum annuum, Capsicum baccatum and Capsicum chinense species. A total of 19 free Apocarotenoids and 8 Apocarotenoids fatty acid esters were identified; β-Apo-8′-carotenal and Apo-8′-zeaxanthinal were also quantified and the β-Apo-8′-carotenal occurrence was in the percentage ranges relative to β-carotene of 0.03–3.87%. PCA was performed as a multivariate display method on the quantified carotenoids and Apocarotenoids, in order to visualize the data structure. Moreover, different e-apoluteinals and 4-oxo-apo-β-carotenals were detected in Capsicum species also for the first time and, to the best of authors knowledge, in any food matrix.
Abdulkerim Eroglu - One of the best experts on this subject based on the ideXlab platform.
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Carotenoid metabolism in mammals, including man: formation, occurrence, and function of Apocarotenoids Thematic Review Series: Fat-Soluble Vitamins: Vitamin A
Journal of Lipid Research, 2013Co-Authors: Abdulkerim Eroglu, Earl H. HarrisonAbstract:Vitamin A was recognized as an essential nutrient 100 years ago. In the 1930s, it became clear that dietary β-carotene was cleaved at its central double to yield vitamin A (retinal or β-apo-15'-carotenal). Thus a great deal of research has focused on the central cleavage of provitamin A carotenoids to form vitamin A (retinoids). The mechanisms of formation and the physiological role(s) of noncentral (eccentric) cleavage of both provitamin A carotenoids and nonprovitamin A carotenoids has been less clear. It is becoming apparent that the Apocarotenoids exert unique biological activities themselves. These compounds are found in the diet and thus may be absorbed in the intestine, or they may form from enzymatic or nonenzymatic cleavage of the parent carotenoids. The mechanism of action of Apocarotenoids in mammals is not fully worked out. However, as detailed in this review, they have profound effects on gene expression and work, at least in part, through the modulation of ligand-activated nuclear receptors. Understanding the interactions of Apocarotenoids with other lipid-binding proteins, chaperones, and metabolizing enzymes will undoubtedly increase our understanding of the biological roles of these carotenoid metabolites.
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Thematic Review Series: Fat-Soluble Vitamins: Vitamin A Carotenoid metabolism in mammals, including man: formation, occurrence, and function of Apocarotenoids
2013Co-Authors: Abdulkerim Eroglu, Earl H. HarrisonAbstract:Vitamin A was recognized as an essential nutrient 100 years ago. In the 1930s, it became clear that dietary - carotene was cleaved at its central double to yield vitamin A (retinal or -apo-15 ' -carotenal). Thus a great deal of research has focused on the central cleavage of provitamin A carote- noids to form vitamin A (retinoids). The mechanisms of for- mation and the physiological role(s) of noncentral (eccentric) cleavage of both provitamin A carotenoids and nonprovitamin A carotenoids has been less clear. It is becoming apparent that the Apocarotenoids exert unique biological activities them- selves. These compounds are found in the diet and thus may be absorbed in the intestine, or they may form from enzymatic or nonenzymatic cleavage of the parent carotenoids. The mechanism of action of Apocarotenoids in mammals is not fully worked out. However, as detailed in this review, they have profound effects on gene expression and work, at least in part, through the modulation of ligand-activated nuclear recep- tors. Understanding the interactions of Apocarotenoids with other lipid-binding proteins, chaperones, and metabolizing en- zymes will undoubtedly increase our understanding of the bio- logical roles of these carotenoid metabolites. —Eroglu, A., and E. H. Harrison. Carotenoid metabolism in mammals, including man: formation, occurrence, and function of Apocarotenoids. J. Lipid Res. 2013. 54: 1719-1730.
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The formation, occurrence, and function of β-Apocarotenoids: β-carotene metabolites that may modulate nuclear receptor signaling
The American Journal of Clinical Nutrition, 2012Co-Authors: Earl H. Harrison, Carlo Dela Seña, Abdulkerim Eroglu, Matthew K. FleshmanAbstract:β-Carotene is the major dietary source of provitamin A. Central cleavage of β-carotene yields 2 molecules of retinal followed by further oxidation to retinoic acid. Eccentric cleavage of β-carotene occurs at double bonds other than the central double bond, and the products of these reactions are β-apocarotenals and β-apocarotenones. We reviewed recent developments in 3 areas: 1): the enzymatic production of β-Apocarotenoids in higher animals; 2) the occurrence of β-Apocarotenoids in foods and animal tissues; and 3) the biological activity of β-Apocarotenoids, particularly on retinoid receptors. HPLC–mass spectrometry techniques were developed to quantify these compounds in mouse serum and tissues and in foods. β-Apo-10′- and -12′-carotenals were detected in mouse serum and liver. β-Apo-8′-, β-apo-10′-, β-apo-12′-, and β-apo-14′-carotenals and β-apo-13-carotenone were detected in orange-fleshed melons. Transactivation assays were performed to see whether Apocarotenoids activate or antagonize retinoid X receptor (RXR) α. Reporter gene constructs and retinoid receptor (RXRα) were transfected into cells, which were used to perform quantitative assays for the activation of this ligand-dependent transcription factor. None of the β-Apocarotenoids significantly activated RXRα. However, β-apo-13-carotenone antagonized the 9-cis-retinoic acid activation of RXRα. Competitive radioligand binding assays showed that this antagonist competes directly with the agonist for binding to purified receptor, a finding confirmed by molecular modeling studies. These findings suggest that a possible biological function of β-Apocarotenoids is their ability to interfere with nuclear receptor signaling. Recent work showed that β-apo-13-carotenone is also a high-affinity antagonist of all 3 retinoic acid receptors (RARα, RARβ, and RARγ).
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Apocarotenoids MODULATE RETINOID RECEPTORS
2012Co-Authors: Abdulkerim ErogluAbstract:s-carotene (BC) is the major dietary source of provitamin A. Central cleavage of BC catalyzed by s-carotene oxygenase 1 yields two molecules of retinaldehyde. Subsequent oxidation produces all-trans-retinoic acid (ATRA) which functions as a ligand for a family of nuclear transcription factors, the retinoic acid receptors (RARs). Eccentric cleavage of BC at non-central double bonds is catalyzed by other enzymes and can also occur non-enzymatically. The products of these reactions are s-apocarotenals and s-apocarotenones, whose biological functions in mammals are unknown. We used reporter gene assays to show that none of the s-Apocarotenoids significantly activated RARs and RXRa. s-Apo-13-carotenone was found to antagonize the activation of RXRa by 9-cis-retinoic acid and was effective at concentrations as low as 1 nM. Molecular modeling studies revealed that s-apo-13-carotenone makes molecular interactions like an antagonist of RXRa. The results suggest a possible function of BACs on RXRa signaling. Moreover, s-apo-14’-carotenal, s-apo-14’-carotenoic acid, and s-apo-13-carotenone antagonized ATRA-induced transactivation of RARs. Competitive radioligand binding assays demonstrated that these putative RAR antagonists compete directly with retinoic acid for high affinity binding to purified receptors. Molecular modeling studies confirmed that s-apo-13-carotenone can directly interact with the ligand binding site of the retinoid receptors. s-Apo-13-carotenone and the s-apo-14’-carotenoids inhibited ATRA-induced expression of retinoid responsive genes in Hep G2 cells. Finally, we developed an LC/MS method and found 3-5 nM s-apo-13-carotenone was present in human plasma. These findings suggest that s-Apocarotenoids function as naturally-occurring retinoid antagonists. We have also tested apo-lycopenoids that have a structural resemblance to s-apo-13-carotenone to see if they exert a similar action as s-apo-13-carotenone in modulating retinoid receptor activation. We found that apo-13-lycopenone was able to block ATRA induced expression of RARs and CYP26A1 like the action of s-apo-13-carotenone. This suggests that the ionone ring may not be prerequisite for s-apo-13-carotenone’s binding to RARs. The antagonism of retinoid signaling by these metabolites may have implications for the activities of dietary s-carotene as a provitamin A and as a modulator of risk for cardiovascular disease and cancer.
Mariosimone Zoccali - One of the best experts on this subject based on the ideXlab platform.
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Determination of free Apocarotenoids and apocarotenoid esters in human colostrum.
Analytical and Bioanalytical Chemistry, 2020Co-Authors: Mariosimone Zoccali, Daniele Giuffrida, Roberta Granese, Fabio Salafia, Paola Dugo, Luigi MondelloAbstract:The presence of carotenoids in human colostrum has been reported in the literature, and xanthophyll esters in human colostrum were recently detected for the first time. However, no published studies have reported whether Apocarotenoids, which are metabolites derived from carotenoid enzymatic or nonenzymatic oxidative cleavage, are present in human colostrum. Therefore, the purpose of the present study was to search for the possible occurrence of Apocarotenoids, including apocarotenoid esters, in human colostrum for the first time by applying an online supercritical fluid extraction–supercritical fluid chromatography–tandem mass spectrometry methodology. Recent evidence related to apocarotenoid transcriptional activity has suggested that they may have beneficial health properties superior to those of their parent carotenoids. Three different Apocarotenoids, namely apo-8′-β-carotenal, apo-8′-lycopenal, and β-citraurin, were identified in intact human colostrum samples, with average concentrations of 85 nmol L−1, 54.6 nmol L−1, and 75.4 nmol L−1, respectively. The overall detection of 16 different free Apocarotenoids and 10 different apocarotenoid fatty acid esters in human colostrum was achieved here for the first time. Their occurrence in human colostrum certainly has implications for newborn health status, since colostrum is the only form of food for the newborn during the very first days of life.
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First Apocarotenoids Profiling of Four Microalgae Strains
Antioxidants, 2019Co-Authors: Mariosimone Zoccali, Daniele Giuffrida, Fabio Salafia, Paola Dugo, Carmen Socaciu, Kari Skjånes, Luigi MondelloAbstract:Both enzymatic or oxidative carotenoids cleavages can often occur in nature and produce a wide range of bioactive Apocarotenoids. Considering that no detailed information is available in the literature regarding the occurrence of Apocarotenoids in microalgae species, the aim of this study was to study the extraction and characterization of Apocarotenoids in four different microalgae strains: Chlamydomonas sp. CCMP 2294, Tetraselmis chuii SAG 8-6, Nannochloropsis gaditana CCMP 526, and Chlorella sorokiniana NIVA-CHL 176. This was done for the first time using an online method coupling supercritical fluid extraction and supercritical fluid chromatography tandem mass spectrometry. A total of 29 different Apocarotenoids, including various apocarotenoid fatty acid esters, were detected: apo-12’-zeaxanthinal, β-apo-12’-carotenal, apo-12-luteinal, and apo-12’-violaxanthal. These were detected in all the investigated strains together with the two apocarotenoid esters, apo-10’-zeaxanthinal-C4:0 and apo-8’-zeaxanthinal-C8:0. The overall extraction and detection time for the Apocarotenoids was less than 10 min, including Apocarotenoids esters, with an overall analysis time of less than 20 min. Moreover, preliminary quantitative data showed that the β-apo-8’-carotenal content was around 0.8% and 2.4% of the parent carotenoid, in the C. sorokiniana and T. chuii strains, respectively. This methodology could be applied as a selective and efficient method for the Apocarotenoids detection.
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Carotenoids and Apocarotenoids determination in intact human blood samples by online supercritical fluid extraction-supercritical fluid chromatography-tandem mass spectrometry.
Analytica Chimica Acta, 2018Co-Authors: Mariosimone Zoccali, Daniele Giuffrida, Fabio Salafia, Salvatore V Giofre, Luigi MondelloAbstract:Abstract A direct on-line method based on the coupling of supercritical fluid extraction and supercritical fluid chromatography with triple quadrupole mass spectrometry detection (SFE-SFC-QqQ/MS) for selected carotenoids determination and Apocarotenoids detection in intact human blood was developed for the first time. Carotenoids and Apocarotenoids were identified by using the available standard together with full scan, selected ion monitoring (SIM), and multiple reaction monitoring (MRM) experiments. Moreover, β-Cryptoxanthin, Zeaxanthin, β-Carotene and Capsanthin were directly quantified by the developed methodology, using a multiple reaction monitoring (MRM) approach; the determined average content of β-carotene was 123.8 nmol L−1 (range 18.7–485.1 nmol L−1), of β-cryptoxanthin was 385.3 nmol L−1 (range 72.5–1920.3 nmol L−1), of zeaxanthin was 396.9 nmol L−1 (range
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Apocarotenoids determination in capsicum chinense jacq cv habanero by supercritical fluid chromatography triple quadrupole mass spectrometry
Food Chemistry, 2017Co-Authors: Daniele Giuffrida, Mariosimone Zoccali, Paola Dugo, Salvatore V Giofre, Luigi MondelloAbstract:Relatively few studies are available in the literature on the Apocarotenoids occurrence in food which are based on liquid chromatographic separation approaches and no reports are available on the Apocarotenoids separation by SFC-APCI (+/-)/QqQ/MS. The dual purpose of this research was to investigate the Apocarotenoids presence in red habanero peppers and to develop a novel SFC-APCI(+/-)/QqQ/MS methodology, using a novel fused-core C30 column, for the Apocarotenoids detection in a fast and efficient way, for the first time. In this study, 25 different Apocarotenoids were identified, 14 were free Apocarotenoids and 11 were Apocarotenoids fatty acids esters. The methodology allowed for all the separations to occur in less then five minutes. The detected Apo-10'-, Apo-14'- and Apo-15-capsorubinals and different Apo-8'-capsorubinal and Apo-10'-zeaxanthinals fatty acid esters had not been previously tentatively identified in any Capsicum species and, to the best of our knowledge, in any food matrix.
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Apocarotenoids profiling in different Capsicum species.
Food Chemistry, 1Co-Authors: Mariosimone Zoccali, Daniele Giuffrida, Fabio Salafia, Paola Dugo, Francesca Rigano, Monica Casale, Luigi MondelloAbstract:Abstract The present study report on the application of an on line supercritical fluid extraction-supercritical fluid chromatography-triple quadrupole/mass spectrometry methodology to the first Apocarotenoids profiling of seventeen different chilli peppers cultivars belonging to Capsicum annuum, Capsicum baccatum and Capsicum chinense species. A total of 19 free Apocarotenoids and 8 Apocarotenoids fatty acid esters were identified; β-Apo-8′-carotenal and Apo-8′-zeaxanthinal were also quantified and the β-Apo-8′-carotenal occurrence was in the percentage ranges relative to β-carotene of 0.03–3.87%. PCA was performed as a multivariate display method on the quantified carotenoids and Apocarotenoids, in order to visualize the data structure. Moreover, different e-apoluteinals and 4-oxo-apo-β-carotenals were detected in Capsicum species also for the first time and, to the best of authors knowledge, in any food matrix.
Oussama Ahrazem - One of the best experts on this subject based on the ideXlab platform.
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efficient production of saffron crocins and picrocrocin in nicotiana benthamiana using a virus driven system
Metabolic Engineering, 2020Co-Authors: Maricarmen Marti, Oussama Ahrazem, Lourdes Gomezgomez, Veronica Aragones, Sarah Frusciante, Gianfranco Diretto, José-antonio DaròsAbstract:Abstract Crocins and picrocrocin are glycosylated Apocarotenoids responsible, respectively, for the color and the unique taste of the saffron spice, known as red gold due to its high price. Several studies have also shown the health-promoting properties of these compounds. However, their high costs hamper the wide use of these metabolites in the pharmaceutical sector. We have developed a virus-driven system to produce remarkable amounts of crocins and picrocrocin in adult Nicotiana benthamiana plants in only two weeks. The system consists of viral clones derived from tobacco etch potyvirus that express specific carotenoid cleavage dioxygenase (CCD) enzymes from Crocus sativus and Buddleja davidii. Metabolic analyses of infected tissues demonstrated that the sole virus-driven expression of C. sativus CsCCD2L or B. davidii BdCCD4.1 resulted in the production of crocins, picrocrocin and safranal. Using the recombinant virus that expressed CsCCD2L, accumulations of 0.2% of crocins and 0.8% of picrocrocin in leaf dry weight were reached in only two weeks. In an attempt to improve apocarotenoid content in N. benthamiana, co-expression of CsCCD2L with other carotenogenic enzymes, such as Pantoea ananatis phytoene synthase (PaCrtB) and saffron β-carotene hydroxylase 2 (BCH2), was performed using the same viral system. This combinatorial approach led to an additional crocin increase up to 0.35% in leaves in which CsCCD2L and PaCrtB were co-expressed. Considering that saffron Apocarotenoids are costly harvested from flower stigma once a year, and that Buddleja spp. flowers accumulate lower amounts, this system may be an attractive alternative for the sustainable production of these appreciated metabolites.
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ugt709g1 a novel uridine diphosphate glycosyltransferase involved in the biosynthesis of picrocrocin the precursor of safranal in saffron crocus sativus
New Phytologist, 2019Co-Authors: Gianfranco Diretto, Oussama Ahrazem, Angela Rubiomoraga, Filippo Sevi, Javier Argandoña, Alessia Fiore, Lourdes GomezgomezAbstract:: Saffron, a spice derived from the dried red stigmas of Crocus sativus, is one of the oldest natural food additives. The flowers have long red stigmas, which store significant quantities of the glycosylated Apocarotenoids crocins and picrocrocin. The apocarotenoid biosynthetic pathway in saffron starts with the oxidative cleavage of zeaxanthin, from which crocins and picrocrocin are derived. In the processed stigmas, picrocrocin is converted to safranal, giving saffron its typical aroma. By a targeted search for differentially expressed uridine diphosphate glycosyltransferases (UGTs) in Crocus transcriptomes, a novel apocarotenoid glucosyltransferase (UGT709G1) from saffron was identified. Biochemical analyses revealed that UGT709G1 showed a high catalytic efficiency toward 2,6,6-trimethyl-4-hydroxy-1-carboxaldehyde-1-cyclohexene (HTCC), making it suited for the biosynthesis of picrocrocin, the precursor of safranal. The role of UGT709G1 in picrocrocin/safranal biosynthesis was supported by the absence or presence of gene expression in a screening for HTCC and picrocrocin production in different Crocus species and by a combined transient expression assay with CsCCD2L in Nicotiana benthamiana leaves. The identification of UGT709G1 completes one of the most highly valued specialized metabolic biosynthetic pathways in plants and provides novel perspectives on the industrial production of picrocrocin to be used as a flavor additive or as a pharmacological constituent.
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Multi-species transcriptome analyses for the regulation of crocins biosynthesis in Crocus
BMC, 2019Co-Authors: Oussama Ahrazem, Javier Argandoña, Andrea Rujas, Angela Rubio-moraga, Raquel Castillo, Alessia Fiore, Lourdes Gómez-gómezAbstract:Abstract Background Crocins are soluble Apocarotenoids that mainly accumulate in the stigma tissue of Crocus sativus and provide the characteristic red color to saffron spice, in addition to being responsible for many of the medicinal properties of saffron. Crocin biosynthesis and accumulation in saffron is developmentally controlled, and the concentration of crocins increases as the stigma develops. Until now, little has been known about the molecular mechanisms governing crocin biosynthesis and accumulation. This study aimed to identify the first set of gene regulatory processes implicated in apocarotenoid biosynthesis and accumulation. Results A large-scale crocin-mediated RNA-seq analysis was performed on saffron and two other Crocus species at two early developmental stages coincident with the initiation of crocin biosynthesis and accumulation. Pairwise comparison of unigene abundance among the samples identified potential regulatory transcription factors (TFs) involved in crocin biosynthesis and accumulation. We found a total of 131 (up- and downregulated) TFs representing a broad range of TF families in the analyzed transcriptomes; by comparison with the transcriptomes from the same developmental stages from other Crocus species, a total of 11 TF were selected as candidate regulators controlling crocin biosynthesis and accumulation. Conclusions Our study generated gene expression profiles of stigmas at two key developmental stages for apocarotenoid accumulation in three different Crocus species. Differential gene expression analyses allowed the identification of transcription factors that provide evidence of environmental and developmental control of the apocarotenoid biosynthetic pathway at the molecular level
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transcriptome analysis in tissue sectors with contrasting crocins accumulation provides novel insights into apocarotenoid biosynthesis and regulation during chromoplast biogenesis
Scientific Reports, 2018Co-Authors: Oussama Ahrazem, Angela Rubiomoraga, Javier Argandoña, Alessia Fiore, Carolina Aguado, Rafael Lujan, Monica Marro, C Araujoandrade, Pablo Lozaalvarez, Gianfranco DirettoAbstract:Crocins, the red soluble Apocarotenoids of saffron, accumulate in the flowers of Crocus species in a developmental and tissue-specific manner. In Crocus sieberi, crocins accumulate in stigmas but also in a distinct yellow tepal sector, which we demonstrate contains chromoplast converted from amyloplasts. Secondary metabolites were analysed by LC-DAD-HRMS, revealing the progressive accumulation of crocetin and crocins in the yellow sector, which were also localized in situ by Raman microspectroscopy. To understand the underlying mechanisms of crocin biosynthesis, we sequenced the C. sieberi tepal transcriptome of two differentially pigmented sectors (yellow and white) at two developmental stages (6 and 8) by Illumina sequencing. A total of 154 million high-quality reads were generated and assembled into 248,099 transcripts. Differentially expressed gene analysis resulted in the identification of several potential candidate genes involved in crocin metabolism and regulation. The results provide a first profile of the molecular events related to the dynamics of crocetin and crocin accumulation during tepal development, and present new information concerning apocarotenoid biosynthesis regulators and their accumulation in Crocus. Further, reveals genes that were previously unknown to affect crocin formation, which could be used to improve crocin accumulation in Crocus plants and the commercial quality of saffron spice.
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evolutionarily distinct carotenoid cleavage dioxygenases are responsible for crocetin production in buddleja davidii
Journal of Experimental Botany, 2017Co-Authors: Oussama Ahrazem, Angela Rubiomoraga, Javier Argandoña, Gianfranco Diretto, Antonio Granell, Jose Manuel Julve, Diego Orzaez, Lourdes GomezgomezAbstract:Crocetin, one of the few colored Apocarotenoids known in nature, is present in flowers and fruits and has long been used medicinally and as a colorant. Saffron is the main source of crocetin, although a few other plants produce lower amounts of this apocarotenoid. Notably, Buddleja davidii accumulates crocetin in its flowers. Recently, a carotenoid dioxygenase cleavage enzyme, CCD2, has been characterized as responsible for crocetin production in Crocus species. We searched for CCD2 homologues in B. davidii and identified several CCD enzymes from the CCD1 and CCD4 subfamilies. Unexpectedly, two out of the three CCD4 enzymes, namely BdCCD4.1 and BdCCD4.3, showed 7,8;7',8' activity in vitro and in vivo over zeaxanthin. In silico analyses of these enzymes and CCD2 allowed the determination of key residues for this activity. Both BdCCD4 genes are highly expressed during flower development and transcripts levels parallel the accumulation of crocins in the petals. Phylogenetic analysis showed that BdCCD4.2 grouped with almost all the characterized CCD4 enzymes, while BdCCD4.1 and BdCCD4.3 form a new sub-cluster together with CCD4 enzymes from certain Lamiales species. The present study indicates that convergent evolution led to the acquisition of 7,8;7',8' apocarotenoid cleavage activity in two separate CCD enzyme families.