Abstract:Capsaicin, produced by diverse Capsicum species, is among the world’s most popular spices and of considerable pharmaceutical relevance. Although the capsaicinoid biosynthetic pathway has been investigated for decades, several biosynthetic steps have remained partly hypothetical. Genetic evidence suggested that the decisive capsaicin synthase is encoded by the Pun1 locus. Yet, the genetic evidence of the Pun1 locus was never corroborated by functionally active capsaicin synthase that presumably catalyzes an ami… Show more
“…A successful attempt of producing the red carotenoid capsanthin in microorganisms was done by overexpressing the CCS gene from C. annuum in Escherichia coli alongside regulating seven other upstream pathway genes . The capsaicin synthase protein purified from a recombinant E. coli host was found to be a monomeric protein of 51 kDa that catalyzed the formation of an amide bond specifically between vanilloylamine and trans-8-methyl-6-nonenoyl-CoA independent of any cofactors . Current biotechnological progress for the industrial scale production of capsaicinoids precursors using genetically engineered organisms, specifically, baker’s yeast ( Saccharomyces cerevisiae ), has been summarized recently …”
Section: Metabolite Engineering For Capsicum Secondary Metabolitesmentioning
Capsicum (chili pepper) is a widely popular and highly consumed fruit crop with beneficial secondary metabolites such as capsaicinoids, carotenoids, flavonoids, and polyphenols, among others. Interestingly, the secondary metabolite profile is a dynamic function of biosynthetic enzymes, regulatory transcription factors, developmental stage, abiotic and biotic environment, and extraction methods. We propose active manipulable genetic, environmental, and extraction controls for the modulation of quality and quantity of desired secondary metabolites in Capsicum species. Specific biosynthetic genes such as Pun (AT3) and AMT in the capsaicinoids pathway and PSY, LCY, and CCS in the carotenoid pathway can be genetically engineered for enhanced production of capsaicinoids and carotenoids, respectively. Generally, secondary metabolites increase with the ripening of the fruit; however, transcriptional regulators such as MYB, bHLH, and ERF control the extent of accumulation in specific tissues. The precise tuning of biotic and abiotic factors such as light, temperature, and chemical elicitors can maximize the accumulation and retention of secondary metabolites in pre-and postharvest settings. Finally, optimized extraction methods such as ultrasonication and supercritical fluid method can lead to a higher yield of secondary metabolites. Together, the integrated understanding of the genetic regulation of biosynthesis, elicitation treatments, and optimization of extraction methods can maximize the industrial production of secondary metabolites in Capsicum.
“…A successful attempt of producing the red carotenoid capsanthin in microorganisms was done by overexpressing the CCS gene from C. annuum in Escherichia coli alongside regulating seven other upstream pathway genes . The capsaicin synthase protein purified from a recombinant E. coli host was found to be a monomeric protein of 51 kDa that catalyzed the formation of an amide bond specifically between vanilloylamine and trans-8-methyl-6-nonenoyl-CoA independent of any cofactors . Current biotechnological progress for the industrial scale production of capsaicinoids precursors using genetically engineered organisms, specifically, baker’s yeast ( Saccharomyces cerevisiae ), has been summarized recently …”
Section: Metabolite Engineering For Capsicum Secondary Metabolitesmentioning
Capsicum (chili pepper) is a widely popular and highly consumed fruit crop with beneficial secondary metabolites such as capsaicinoids, carotenoids, flavonoids, and polyphenols, among others. Interestingly, the secondary metabolite profile is a dynamic function of biosynthetic enzymes, regulatory transcription factors, developmental stage, abiotic and biotic environment, and extraction methods. We propose active manipulable genetic, environmental, and extraction controls for the modulation of quality and quantity of desired secondary metabolites in Capsicum species. Specific biosynthetic genes such as Pun (AT3) and AMT in the capsaicinoids pathway and PSY, LCY, and CCS in the carotenoid pathway can be genetically engineered for enhanced production of capsaicinoids and carotenoids, respectively. Generally, secondary metabolites increase with the ripening of the fruit; however, transcriptional regulators such as MYB, bHLH, and ERF control the extent of accumulation in specific tissues. The precise tuning of biotic and abiotic factors such as light, temperature, and chemical elicitors can maximize the accumulation and retention of secondary metabolites in pre-and postharvest settings. Finally, optimized extraction methods such as ultrasonication and supercritical fluid method can lead to a higher yield of secondary metabolites. Together, the integrated understanding of the genetic regulation of biosynthesis, elicitation treatments, and optimization of extraction methods can maximize the industrial production of secondary metabolites in Capsicum.
“…Die Technologie der Sequenzierung ist so weit vorangeschritten, dass zahlreiche pflanzliche Genome wie das von Paprika bekannt sind und die transkribierte RNA einzelner Gene zur Herstellung der entsprechenden Enzyme in Bakterien oder Hefen genutzt werden kann. So können unvollständig oder nicht verstandene enzymatische Schritte identifiziert und die Eigenschaften der Enzyme charakterisiert werden [2] – wie erst kürzlich im Fall der Piperinsynthase, die genau wie die Capsaicinsynthase eine Amidbindung knüpfen kann. Wie Capsaicin bindet auch das strukturell ähnliche Piperin an den TRPV1‐Rezeptor und daher bewirkt auch diese Substanz einen scharfen Geschmack.…”
Section: Enzyme Vermitteln Die Amidbindungunclassified
“…Nun gelang es, auch den letzten Reaktionsschritt der Biosynthese von Capsaicin aus der Chilischote (Capsicum spec.) aufzuklären und erstmals die enzymatische Aktivität der lange gesuchten Capsaicinsynthase nachzuweisen [2]. (Fotos: Thomas Vogt, Leibniz‐Institut für Pflanzenbiochemie, Halle) Formel Vanilloylamin korrigiert am 25.07.2023…”
Section: Enzyme Vermitteln Die Amidbindungunclassified
“…Im Fall der Capsaicinsynthase wurden von den Autoren bereits veröffentlichte Gensequenzen synthetisch optimiert und auf einem Plasmid in einen speziellen E. coli‐Stamm eingeschleust, der für die Produktion von BAHD‐Acyltransferasen geeignet erschien [2]. Der komplette Proteinextrakt des Bakteriums wurde nachfolgend gereinigt und Sequenzstücke der Capsaicinsynthase mittels Massenspektrometrie in diesem Extrakt nachgewiesen.…”
Section: Vom Gen Zum Aktiven Enzymunclassified
“…Im Fall der Capsaicinsynthase wur den von den Autoren bereits ver öffentlichte Gensequenzen syn thetisch optimiert und auf einem Plasmid in einen speziellen E. coli Stamm eingeschleust, der für die Produktion von BAHDAcyltransfera sen geeignet erschien [2]…”
Wenn mit Chili gewürzte Speisen im Mund brennen, ist das Warum klar: Capsaicin fand den Weg zu seinen Rezeptoren. Doch in der pflanzlichen Biosynthese war noch unklar, wie spezifisch eines der Schlüsselenzyme ist und ob es mit oder ohne Cofaktoren arbeitet. Nun konnte die Capsaicinsynthase rekombinant hergestellt und auf enzymatischer Ebene näher untersucht werden.
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