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Susheel Kumar Sharma - One of the best experts on this subject based on the ideXlab platform.
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Efficient enhancement of Capsaicinoids biosynthesis in cell suspension cultures of Capsicum chinense Jacq. cv. ‘Umorok’ by elicitors and differential gene expression analysis of elicited cultures
Plant Cell Tissue and Organ Culture (PCTOC), 2020Co-Authors: Khaidem Chanu Kabita, K. Sanatombi, Susheel Kumar SharmaAbstract:Capsaicinoids are responsible for pungency in chillies and have multiple health benefits and industrial applications. In vitro cell cultures of Capsicum with high Capsaicinoids content may have the potential to serve as an alternative source for production of Capsaicinoids. The present study investigated the effects of various elicitors, precursors and intermediates on the production of Capsaicinoids in cell suspension cultures established from callus derived from the placental tissue explants of the highly pungent chilli cultivar, Capsicum chinense Jacq. cv. ‘Umorok’. The study also, for the first time, reports the effect of elicitors on expression patterns of Capsaicinoids biosynthesis genes in cell suspension cultures of C. chinense by quantitative real-time PCR. The maximum Capsaicinoids content was obtained in cells elicitated with chitosan with average capsaicin and dihydrocapsaicin contents of 2.87 mg g^−1 fresh weight (FW) and 1.03 mg g^−1 FW, respectively. The expressions of the studied candidate genes were significantly up-regulated in response to elicitation. Enhancement of Capsaicinoids biosynthesis by elicitation with chitosan was found to be associated with the up-regulation of the expression of all the studied candidate genes. The present study shows the association of different enhancement potential of different elicitors with differential expression pattern of the candidate genes of Capsaicinoids biosynthesis pathways in cell suspension cultures of C. chinense and the information provided may help in understanding the key mechanism for enhancement of Capsaicinoids biosynthesis by elicitation. The present study reports the change in gene expression profiles of capsaicinoid biosynthesis pathway genes in response to elicitation in cell suspension cultures of Capsicum chinense Jacq.
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Analysis of capsaicinoid biosynthesis pathway genes expression in callus cultures of Capsicum chinense Jacq. cv. ‘Umorok’
Plant Cell Tissue and Organ Culture (PCTOC), 2019Co-Authors: Khaidem Chanu Kabita, Susheel Kumar Sharma, K. SanatombiAbstract:The placental tissue of the highly pungent chilli cultivar, Capsicum chinense Jacq. cv. ‘Umorok’, is used as explants for callus induction. Callus cultures were subcultured after every 32 days and growth curves for a period of six consecutive growth cycles were studied till a stable Capsaicinoids producing callus cultures were obtained. The Capsaicinoids content in placental tissue explants decreased gradually during the first 2 months of culture as the explants dedifferentiated to form friable callus while the biomass and capsaicinoid content did not show much change in the subsequent growth cycles. The maximum callus biomass of 7.8 g freshweight (FW) or 0.56 g dry weight (DW) per culture were obtained on the 24th day of every growth cycle and the maximum average Capsaicinoids content (1.6 mg g^−1 FW capsaicin and 0.78 mg g^−1 FW dihydrocapsaicin) were obtained on the 20th day of every growth cycle. To investigate the underlying dynamics for capsaicinoid biosynthesis during callus formation, comparative gene expression analysis of the genes involved in capsaicinoid biosynthesis pathway were also studied by qRT-PCR analysis. When compared with placental tissue, all the studied genes showed reduced expression during callus formation, especially putative aminotransferase (pAMT) and pungent gene 1 (Pun1), which were extensively down regulated from the 3rd month onwards in the callus cultures. Therefore, the present study revealed that the down-regulated expression of mainly two putative genes in capsaicinoid biosynthetic pathway (pAMT and Pun1) resulted in lower accumulation of Capsaicinoids in callus cultures compared to placental tissues of fruits. The present study reports the gene expression analysis of the genes involved in capsaicinoid biosynthesis pathway during callus formation from placental tissue explants of Capsicum chinense Jacq.
P A Biacs - One of the best experts on this subject based on the ideXlab platform.
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supercritical co2 and subcritical propane extraction of pungent paprika and quantification of carotenoids tocopherols and Capsaicinoids
Journal of Agricultural and Food Chemistry, 2001Co-Authors: M H Gnayfeed, Vendel Illes, Hussein G Daood, P A BiacsAbstract:Ground paprika (Capsicum annuum L.) was extracted with supercritical carbon dioxide (SC-CO2) and subcritical propane at different conditions of pressure and temperature to estimate the yield and variation in carotenoid, tocopherol, and capsaicinoid contents and composition. The yield of paprika extract was found to be affected by the extraction conditions with SC-CO2 but fairly constant at different conditions with subcritical propane. The maximum yields of oleoresin were 7.9 and 8.1% of ground paprika by SC-CO2 and subcritical propane, respectively. The quantitative distribution of carotenoids, tocopherols, and Capsaicinoids between paprika extract and powder was influenced by extraction conditions. SC-CO2 was inefficient in the extraction of diesters of xanthophylls even at 400 bar and 55 °C, whereas tocopherols and Capsaicinoids were easy to extract at these conditions. Under mild conditions subcritical propane was superior to SC-CO2 in the extraction of carotenoids and tocopherols but less efficient i...
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supercritical co2 and subcritical propane extraction of pungent paprika and quantification of carotenoids tocopherols and Capsaicinoids
Journal of Agricultural and Food Chemistry, 2001Co-Authors: M H Gnayfeed, Vendel Illes, Hussein G Daood, P A BiacsAbstract:Ground paprika (Capsicum annuum L.) was extracted with supercritical carbon dioxide (SC-CO(2)) and subcritical propane at different conditions of pressure and temperature to estimate the yield and variation in carotenoid, tocopherol, and capsaicinoid contents and composition. The yield of paprika extract was found to be affected by the extraction conditions with SC-CO(2) but fairly constant at different conditions with subcritical propane. The maximum yields of oleoresin were 7.9 and 8.1% of ground paprika by SC-CO(2) and subcritical propane, respectively. The quantitative distribution of carotenoids, tocopherols, and Capsaicinoids between paprika extract and powder was influenced by extraction conditions. SC-CO(2) was inefficient in the extraction of diesters of xanthophylls even at 400 bar and 55 degrees C, whereas tocopherols and Capsaicinoids were easy to extract at these conditions. Under mild conditions subcritical propane was superior to SC-CO(2) in the extraction of carotenoids and tocopherols but less efficient in the extraction of Capsaicinoids.
K. Sanatombi - One of the best experts on this subject based on the ideXlab platform.
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Efficient enhancement of Capsaicinoids biosynthesis in cell suspension cultures of Capsicum chinense Jacq. cv. ‘Umorok’ by elicitors and differential gene expression analysis of elicited cultures
Plant Cell Tissue and Organ Culture (PCTOC), 2020Co-Authors: Khaidem Chanu Kabita, K. Sanatombi, Susheel Kumar SharmaAbstract:Capsaicinoids are responsible for pungency in chillies and have multiple health benefits and industrial applications. In vitro cell cultures of Capsicum with high Capsaicinoids content may have the potential to serve as an alternative source for production of Capsaicinoids. The present study investigated the effects of various elicitors, precursors and intermediates on the production of Capsaicinoids in cell suspension cultures established from callus derived from the placental tissue explants of the highly pungent chilli cultivar, Capsicum chinense Jacq. cv. ‘Umorok’. The study also, for the first time, reports the effect of elicitors on expression patterns of Capsaicinoids biosynthesis genes in cell suspension cultures of C. chinense by quantitative real-time PCR. The maximum Capsaicinoids content was obtained in cells elicitated with chitosan with average capsaicin and dihydrocapsaicin contents of 2.87 mg g^−1 fresh weight (FW) and 1.03 mg g^−1 FW, respectively. The expressions of the studied candidate genes were significantly up-regulated in response to elicitation. Enhancement of Capsaicinoids biosynthesis by elicitation with chitosan was found to be associated with the up-regulation of the expression of all the studied candidate genes. The present study shows the association of different enhancement potential of different elicitors with differential expression pattern of the candidate genes of Capsaicinoids biosynthesis pathways in cell suspension cultures of C. chinense and the information provided may help in understanding the key mechanism for enhancement of Capsaicinoids biosynthesis by elicitation. The present study reports the change in gene expression profiles of capsaicinoid biosynthesis pathway genes in response to elicitation in cell suspension cultures of Capsicum chinense Jacq.
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Analysis of capsaicinoid biosynthesis pathway genes expression in callus cultures of Capsicum chinense Jacq. cv. ‘Umorok’
Plant Cell Tissue and Organ Culture (PCTOC), 2019Co-Authors: Khaidem Chanu Kabita, Susheel Kumar Sharma, K. SanatombiAbstract:The placental tissue of the highly pungent chilli cultivar, Capsicum chinense Jacq. cv. ‘Umorok’, is used as explants for callus induction. Callus cultures were subcultured after every 32 days and growth curves for a period of six consecutive growth cycles were studied till a stable Capsaicinoids producing callus cultures were obtained. The Capsaicinoids content in placental tissue explants decreased gradually during the first 2 months of culture as the explants dedifferentiated to form friable callus while the biomass and capsaicinoid content did not show much change in the subsequent growth cycles. The maximum callus biomass of 7.8 g freshweight (FW) or 0.56 g dry weight (DW) per culture were obtained on the 24th day of every growth cycle and the maximum average Capsaicinoids content (1.6 mg g^−1 FW capsaicin and 0.78 mg g^−1 FW dihydrocapsaicin) were obtained on the 20th day of every growth cycle. To investigate the underlying dynamics for capsaicinoid biosynthesis during callus formation, comparative gene expression analysis of the genes involved in capsaicinoid biosynthesis pathway were also studied by qRT-PCR analysis. When compared with placental tissue, all the studied genes showed reduced expression during callus formation, especially putative aminotransferase (pAMT) and pungent gene 1 (Pun1), which were extensively down regulated from the 3rd month onwards in the callus cultures. Therefore, the present study revealed that the down-regulated expression of mainly two putative genes in capsaicinoid biosynthetic pathway (pAMT and Pun1) resulted in lower accumulation of Capsaicinoids in callus cultures compared to placental tissues of fruits. The present study reports the gene expression analysis of the genes involved in capsaicinoid biosynthesis pathway during callus formation from placental tissue explants of Capsicum chinense Jacq.
Khaidem Chanu Kabita - One of the best experts on this subject based on the ideXlab platform.
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Efficient enhancement of Capsaicinoids biosynthesis in cell suspension cultures of Capsicum chinense Jacq. cv. ‘Umorok’ by elicitors and differential gene expression analysis of elicited cultures
Plant Cell Tissue and Organ Culture (PCTOC), 2020Co-Authors: Khaidem Chanu Kabita, K. Sanatombi, Susheel Kumar SharmaAbstract:Capsaicinoids are responsible for pungency in chillies and have multiple health benefits and industrial applications. In vitro cell cultures of Capsicum with high Capsaicinoids content may have the potential to serve as an alternative source for production of Capsaicinoids. The present study investigated the effects of various elicitors, precursors and intermediates on the production of Capsaicinoids in cell suspension cultures established from callus derived from the placental tissue explants of the highly pungent chilli cultivar, Capsicum chinense Jacq. cv. ‘Umorok’. The study also, for the first time, reports the effect of elicitors on expression patterns of Capsaicinoids biosynthesis genes in cell suspension cultures of C. chinense by quantitative real-time PCR. The maximum Capsaicinoids content was obtained in cells elicitated with chitosan with average capsaicin and dihydrocapsaicin contents of 2.87 mg g^−1 fresh weight (FW) and 1.03 mg g^−1 FW, respectively. The expressions of the studied candidate genes were significantly up-regulated in response to elicitation. Enhancement of Capsaicinoids biosynthesis by elicitation with chitosan was found to be associated with the up-regulation of the expression of all the studied candidate genes. The present study shows the association of different enhancement potential of different elicitors with differential expression pattern of the candidate genes of Capsaicinoids biosynthesis pathways in cell suspension cultures of C. chinense and the information provided may help in understanding the key mechanism for enhancement of Capsaicinoids biosynthesis by elicitation. The present study reports the change in gene expression profiles of capsaicinoid biosynthesis pathway genes in response to elicitation in cell suspension cultures of Capsicum chinense Jacq.
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Analysis of capsaicinoid biosynthesis pathway genes expression in callus cultures of Capsicum chinense Jacq. cv. ‘Umorok’
Plant Cell Tissue and Organ Culture (PCTOC), 2019Co-Authors: Khaidem Chanu Kabita, Susheel Kumar Sharma, K. SanatombiAbstract:The placental tissue of the highly pungent chilli cultivar, Capsicum chinense Jacq. cv. ‘Umorok’, is used as explants for callus induction. Callus cultures were subcultured after every 32 days and growth curves for a period of six consecutive growth cycles were studied till a stable Capsaicinoids producing callus cultures were obtained. The Capsaicinoids content in placental tissue explants decreased gradually during the first 2 months of culture as the explants dedifferentiated to form friable callus while the biomass and capsaicinoid content did not show much change in the subsequent growth cycles. The maximum callus biomass of 7.8 g freshweight (FW) or 0.56 g dry weight (DW) per culture were obtained on the 24th day of every growth cycle and the maximum average Capsaicinoids content (1.6 mg g^−1 FW capsaicin and 0.78 mg g^−1 FW dihydrocapsaicin) were obtained on the 20th day of every growth cycle. To investigate the underlying dynamics for capsaicinoid biosynthesis during callus formation, comparative gene expression analysis of the genes involved in capsaicinoid biosynthesis pathway were also studied by qRT-PCR analysis. When compared with placental tissue, all the studied genes showed reduced expression during callus formation, especially putative aminotransferase (pAMT) and pungent gene 1 (Pun1), which were extensively down regulated from the 3rd month onwards in the callus cultures. Therefore, the present study revealed that the down-regulated expression of mainly two putative genes in capsaicinoid biosynthetic pathway (pAMT and Pun1) resulted in lower accumulation of Capsaicinoids in callus cultures compared to placental tissues of fruits. The present study reports the gene expression analysis of the genes involved in capsaicinoid biosynthesis pathway during callus formation from placental tissue explants of Capsicum chinense Jacq.
Christopher A Reilly - One of the best experts on this subject based on the ideXlab platform.
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Cytochrome P450-Dependent Modification of Capsaicinoids: Pharmacological Inactivation and Bioactivation Mechanisms
Role of Capsaicin in Oxidative Stress and Cancer, 2013Co-Authors: Christopher A ReillyAbstract:Capsaicinoids are the chemicals that make chili peppers “hot.” Human exposure to Capsaicinoids through their diet is frequent and often considerable. Capsaicinoids are also used in a variety of consumer products, nutraceutical, and pharmaceutical preparations, including pepper spray self-defense products, supplements for weight loss, and therapeutics for pain management, and antioxidant defense/cancer chemoprevention. In the human body, Capsaicinoids are rapidly degraded and/or modified by a variety of enzymatic processes that significantly alter the pharmacological and toxicological properties of the Capsaicinoids. This chapter provides as review of the enzymatic processes that appear to be critical in the clearance of Capsaicinoids from cells in the human body, with an emphasis on Cytochrome P450-dependent processes. Additionally, the consequences of P450-mediated capsaicinoid metabolism with respect to the biological activity of Capsaicinoids as dietary supplements and therapeutic molecules are discussed.
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assessment of pepper spray product potency in asian and caucasian forearm skin using transepidermal water loss skin temperature and reflectance colorimetry
Journal of Applied Toxicology, 2006Co-Authors: Lynn K Pershing, Judy L Corlett, Christopher A Reilly, Dennis J CrouchAbstract:Historically, pepper spray product potency has been established using a taste test evaluation. A taste test is subjective and may not be appropriate for assessing pepper potency in skin. The current study evaluated chemically diverse pepper sprays in human forearm skin using three objective, noninvasive parameters: transepidermal water loss, skin surface temperature and erythema, as a means for assessing dermal pharmacology, toxicology and product potency. Five commercial pepper spray products containing various capsaicinoid analogs at various concentrations were evaluated in duplicate on volar forearms of six Caucasians and six Asians using a 10 min exposure. Mean surface skin temperature, transepidermal water loss results were highly variable and therefore did not demonstrate dose responsive behavior to increasing capsaicinoid concentrations. Erythema, as measured by increases in a* (reflected light in the red-to-green color spectrum) of the L*a*b* uniform color scale, was superior among parameters evaluated in discriminating pepper spray potency and correlated well with the relative and total capsaicinoid concentration in the products. Products containing greater than 16 mg ml(-1) capsaicinoid concentration produced greater erythema responses in Caucasians than Asians. Asians responded greater to the synthetic analog, nonivamide, than to mixtures of Capsaicinoids, while Caucasians responded equally to both capsaicinoid analogs. Thus, pepper spray product potency in human skin reflects the total capsaicinoid concentration, the specific capsaicin analog(s) present, and the race of the individual exposed. The finding that the reflectance colorimeter a* scale can differentiate these parameters in skin will have a significant impact on evaluating the use and efficacy of pepper spray products in humans. Language: en
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Metabolism of Capsaicinoids by P450 enzymes: a review of recent findings on reaction mechanisms, bio-activation, and detoxification processes.
Drug metabolism reviews, 2006Co-Authors: Christopher A Reilly, Garold S. YostAbstract:Capsaicinoids are botanical irritants present in chili peppers. Chili pepper extracts and Capsaicinoids are common dietary constituents and important pharmaceutical agents. Use of these substances in modern consumer products and medicinal preparations occurs worldwide. Capsaicinoids are the principals of pepper spray self-defense weapons and several over-the-counter pain treatments as well as the active component of many dietary supplements. Capsaicinoids interact with the capsaicin receptor (a.k.a., VR1 or TRPV1) to produce acute pain and cough as well as long-term analgesia. Capsaicinoids are also toxic to many cells via TRPV1-dependent and independent mechanisms. Chemical modifications to Capsaicinoids by P450 enzymes decreases their potency at TRPV1 and reduces the pharmacological and toxicological phenomena associated with TRPV1 stimulation. Metabolism of Capsaicinoids by P450 enzymes also produces reactive electrophiles capable of modifying biological macromolecules. This review highlights data describing specific mechanisms by which P450 enzymes convert the Capsaicinoids to novel products and explores the relationship between capsaicinoid metabolism and its effects on capsaicinoid pharmacology and toxicology.
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structural and enzymatic parameters that determine alkyl dehydrogenation hydroxylation of Capsaicinoids by cytochrome p450 enzymes
Drug Metabolism and Disposition, 2005Co-Authors: Christopher A Reilly, Garold S. YostAbstract:Previous studies on the metabolism of Capsaicinoids, natural products isolated from chili peppers, demonstrated the production of unique macrocyclic, alkyl dehydrogenated, ω-, and ω-1-hydroxylated products. This study investigated the structural and enzymatic parameters that direct selective alkyl dehydrogenation and hydroxylation of Capsaicinoids, using a variety of structurally related capsaicinoid analogs and cytochrome P450 (P450) enzymes. CYP2C9 preferentially catalyzed alkyl dehydrogenation, whereas CYP2E1 and 3A4 catalyzed ω- and ω-1-hydroxylation, respectively. Analysis of incubations containing various P450s and structural variants of capsaicin by liquid chromatography-tandem mass spectrometry demonstrated similarities in the rate of capsaicinoid metabolism, but marked differences in the metabolite profiles. Production of macrocyclic and ω-1-hydroxylated metabolites from the various Capsaicinoids was dependent on the structure of the alkyl terminus and P450 enzyme. A tertiary carbon at the ω-1 position, coupled to an adjacent unsaturated bond at the ω-2,3 position, enhanced the formation of the macrocyclic and dehydrogenated metabolites and were requisite structural features for ω-1-hydroxylated product formation. Conversely, substrates lacking these structural features were efficiently oxidized to the ω-hydroxylated metabolite. These data were consistent with our hypothesis that metabolism of the alkyl portion of Capsaicinoids was governed, in part, by the stability and propensity to form an intermediate radical and a carbocation, and a direct interaction between the alkyl terminus and the heme of many P450 enzymes. These results provided valuable insights into potential mechanisms by which P450s metabolize Capsaicinoids and highlight critical chemical features that may also govern the metabolism of structurally related compounds including fatty acids, monoter-penes, and isoprenoids.
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effects of vehicle on the uptake and elimination kinetics of Capsaicinoids in human skin in vivo
Toxicology and Applied Pharmacology, 2004Co-Authors: Lynn K Pershing, Judy L Corlett, Christopher A Reilly, Dennis J CrouchAbstract:While the physiologic and molecular effects of Capsaicinoids have been extensively studied in various model systems by a variety of administration routes, little is known about the uptake and elimination kinetic profiles in human skin following topical exposure. The present study evaluated the uptake and elimination kinetics of Capsaicinoids in human stratum corneum following a single topical exposure to 3% solutions containing 55% capsaicin, 35% dihydrocapsaicin, and 10% other analogues prepared in three vehicles: mineral oil (MO), propylene glycol (PG), and isopropyl alcohol (IPA). Capsaicinoid solutions were evaluated simultaneously in a random application pattern on the volar forearms of 12 subjects using a small, single 150-microg dose. Capsaicin and dihydrocapsaicin were recovered from human skin using commercial adhesive discs to harvest stratum corneum from treated sites. Capsaicinoids were extracted from the stratum corneum-adhesive discs and quantified by liquid chromatography/mass spectroscopy (LC/MS). Both Capsaicinoids were detected in stratum corneum 1 min after application with all vehicles and achieved a pseudo-steady state shortly thereafter. IPA delivered three times greater capsaicin and dihydrocapsaicin into the human stratum corneum than PG or MO at all time points investigated. The Cmax of capsaicin in IPA, PG, and MO was 16.1, 6.2, and 6.5 microg, respectively. The dihydrocapsaicin content was 60% of capsaicin with all vehicles. The estimated T(half) of capsaicin and dihydrocapsaicin in the three vehicles was similar (24 h). Thus, maximal cutaneous capsaicinoid concentrations were achieved quickly in the human stratum corneum and were concentration and vehicle dependent. In contrast, capsaicinoid half-life was long and vehicle independent.