Identifying Bixa orellana L. new carotenoid cleavage dioxygenases 1 and 4 potentially involved in bixin biosynthesis
Carotene cleavage dioxygenases (CCDs) are a large family of Fe2+ dependent enzymes responsible for the production of a wide variety of apocarotenoids, such as bixin. Among the natural apocarotenoids, bixin is second in economic importance. It has a red-orange color and is produced mainly in the seeds of B. orellana. The biosynthesis of bixin aldehyde from the oxidative cleavage of lycopene at 5,6/5′,6′ bonds by a CCD is considered the first step of bixin biosynthesis. Eight BoCCD (BoCCD1-1, BoCCD1-3, BoCCD1-4, CCD4-1, BoCCD4-2, BoCCD4-3 and BoCCD4-4) genes potentially involved in the first step of B. orellana bixin biosynthesis have been identified. However, the cleavage activity upon lycopene to produce bixin aldehyde has only been demonstrated for BoCCD1-1 and BoCCD4-3. Using in vivo (Escherichia coli) and in vitro approaches, we determined that the other identified BoCCDs enzymes (BoCCD1-3, BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4) also participate in the biosynthesis of bixin aldehyde from lycopene. The LC-ESI-QTOF-MS/MS analysis showed a peak corresponding to bixin aldehyde (m/z 349.1) in pACCRT-EIB E. coli cells that express the BoCCD1 and BoCCD4 proteins, which was confirmed by in vitro enzymatic assay. Interestingly, in the in vivo assay of BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4, bixin aldehyde was oxidized to norbixin (m/z 380.2), the second product of the bixin biosynthesis pathway. In silico analysis also showed that BoCCD1 and BoCCD4 proteins encode functional dioxygenases that can use lycopene as substrate. The production of bixin aldehyde and norbixin was corroborated based on their ion fragmentation pattern, as well as by Fourier transform infrared (FTIR) spectroscopy. This work made it possible to clarify at the same time the first and second steps of the bixin biosynthesis pathway that had not been evaluated for a long time. Copyright © 2022 Us-Camas, Aguilar-Espinosa, Rodríguez-Campos, Vallejo-Cardona, Carballo-Uicab, Serrano-Posada and Rivera-Madrid.
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Subjects: | info:eu-repo/classification/Autores/BoCCD1, info:eu-repo/classification/Autores/BoCCD4, info:eu-repo/classification/Autores/BIXIN ALDEHYDE, info:eu-repo/classification/Autores/NORBIXIN, info:eu-repo/classification/Autores/BIXIN BIOSYNTHESIS, info:eu-repo/classification/Autores/BIXA ORELLANA L., info:eu-repo/classification/Autores/APOCAROTENOIDS, info:eu-repo/classification/Autores/CAROTENOIDS, info:eu-repo/classification/cti/2, info:eu-repo/classification/cti/24, info:eu-repo/classification/cti/2415, info:eu-repo/classification/cti/241502, |
Online Access: | http://cicy.repositorioinstitucional.mx/jspui/handle/1003/2815 |
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dig-cicy-1003-28152023-10-24T19:28:38Z Identifying Bixa orellana L. new carotenoid cleavage dioxygenases 1 and 4 potentially involved in bixin biosynthesis ROSA YAZMIN US CAMAS Margarita de Lourdes Aguilar Espinosa Jacobo Rodríguez Campos Alba Adriana Vallejo Cardona Víctor Manuel Carballo Uicab HUGO JAVIER SERRANO POSADA Renata Lourdes Bárbara Rivera Madrid 2022 info:eu-repo/semantics/article Carotene cleavage dioxygenases (CCDs) are a large family of Fe2+ dependent enzymes responsible for the production of a wide variety of apocarotenoids, such as bixin. Among the natural apocarotenoids, bixin is second in economic importance. It has a red-orange color and is produced mainly in the seeds of B. orellana. The biosynthesis of bixin aldehyde from the oxidative cleavage of lycopene at 5,6/5′,6′ bonds by a CCD is considered the first step of bixin biosynthesis. Eight BoCCD (BoCCD1-1, BoCCD1-3, BoCCD1-4, CCD4-1, BoCCD4-2, BoCCD4-3 and BoCCD4-4) genes potentially involved in the first step of B. orellana bixin biosynthesis have been identified. However, the cleavage activity upon lycopene to produce bixin aldehyde has only been demonstrated for BoCCD1-1 and BoCCD4-3. Using in vivo (Escherichia coli) and in vitro approaches, we determined that the other identified BoCCDs enzymes (BoCCD1-3, BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4) also participate in the biosynthesis of bixin aldehyde from lycopene. The LC-ESI-QTOF-MS/MS analysis showed a peak corresponding to bixin aldehyde (m/z 349.1) in pACCRT-EIB E. coli cells that express the BoCCD1 and BoCCD4 proteins, which was confirmed by in vitro enzymatic assay. Interestingly, in the in vivo assay of BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4, bixin aldehyde was oxidized to norbixin (m/z 380.2), the second product of the bixin biosynthesis pathway. In silico analysis also showed that BoCCD1 and BoCCD4 proteins encode functional dioxygenases that can use lycopene as substrate. The production of bixin aldehyde and norbixin was corroborated based on their ion fragmentation pattern, as well as by Fourier transform infrared (FTIR) spectroscopy. This work made it possible to clarify at the same time the first and second steps of the bixin biosynthesis pathway that had not been evaluated for a long time. Copyright © 2022 Us-Camas, Aguilar-Espinosa, Rodríguez-Campos, Vallejo-Cardona, Carballo-Uicab, Serrano-Posada and Rivera-Madrid. info:eu-repo/classification/Autores/BoCCD1 info:eu-repo/classification/Autores/BoCCD4 info:eu-repo/classification/Autores/BIXIN ALDEHYDE info:eu-repo/classification/Autores/NORBIXIN info:eu-repo/classification/Autores/BIXIN BIOSYNTHESIS info:eu-repo/classification/Autores/BIXA ORELLANA L. info:eu-repo/classification/Autores/APOCAROTENOIDS info:eu-repo/classification/Autores/CAROTENOIDS info:eu-repo/classification/cti/2 info:eu-repo/classification/cti/24 info:eu-repo/classification/cti/2415 info:eu-repo/classification/cti/241502 info:eu-repo/classification/cti/241502 Frontiers in Plant Science, 13, 224, 2022. http://cicy.repositorioinstitucional.mx/jspui/handle/1003/2815 info:eu-repo/semantics/datasetDOI/doi: 10.3389/fpls.2022.829089 info:eu-repo/semantics/openAccess eng citation:Us-Camas R, Aguilar-Espinosa M, Rodríguez-Campos J, Vallejo-Cardona AA, Carballo-Uicab VM, Serrano-Posada H and Rivera-Madrid R (2022) Identifying Bixa orellana L. New Carotenoid Cleavage Dioxygenases 1 and 4 Potentially Involved in Bixin Biosynthesis. Front. Plant Sci. 13:829089. doi:10.3389/fpls.2022.829089 http://creativecommons.org/licenses/by-nc-nd/4.0 info:eu-repo/semantics/publishedVersion application/pdf |
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info:eu-repo/classification/Autores/BoCCD1 info:eu-repo/classification/Autores/BoCCD4 info:eu-repo/classification/Autores/BIXIN ALDEHYDE info:eu-repo/classification/Autores/NORBIXIN info:eu-repo/classification/Autores/BIXIN BIOSYNTHESIS info:eu-repo/classification/Autores/BIXA ORELLANA L. info:eu-repo/classification/Autores/APOCAROTENOIDS info:eu-repo/classification/Autores/CAROTENOIDS info:eu-repo/classification/cti/2 info:eu-repo/classification/cti/24 info:eu-repo/classification/cti/2415 info:eu-repo/classification/cti/241502 info:eu-repo/classification/cti/241502 info:eu-repo/classification/Autores/BoCCD1 info:eu-repo/classification/Autores/BoCCD4 info:eu-repo/classification/Autores/BIXIN ALDEHYDE info:eu-repo/classification/Autores/NORBIXIN info:eu-repo/classification/Autores/BIXIN BIOSYNTHESIS info:eu-repo/classification/Autores/BIXA ORELLANA L. info:eu-repo/classification/Autores/APOCAROTENOIDS info:eu-repo/classification/Autores/CAROTENOIDS info:eu-repo/classification/cti/2 info:eu-repo/classification/cti/24 info:eu-repo/classification/cti/2415 info:eu-repo/classification/cti/241502 info:eu-repo/classification/cti/241502 |
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info:eu-repo/classification/Autores/BoCCD1 info:eu-repo/classification/Autores/BoCCD4 info:eu-repo/classification/Autores/BIXIN ALDEHYDE info:eu-repo/classification/Autores/NORBIXIN info:eu-repo/classification/Autores/BIXIN BIOSYNTHESIS info:eu-repo/classification/Autores/BIXA ORELLANA L. info:eu-repo/classification/Autores/APOCAROTENOIDS info:eu-repo/classification/Autores/CAROTENOIDS info:eu-repo/classification/cti/2 info:eu-repo/classification/cti/24 info:eu-repo/classification/cti/2415 info:eu-repo/classification/cti/241502 info:eu-repo/classification/cti/241502 info:eu-repo/classification/Autores/BoCCD1 info:eu-repo/classification/Autores/BoCCD4 info:eu-repo/classification/Autores/BIXIN ALDEHYDE info:eu-repo/classification/Autores/NORBIXIN info:eu-repo/classification/Autores/BIXIN BIOSYNTHESIS info:eu-repo/classification/Autores/BIXA ORELLANA L. info:eu-repo/classification/Autores/APOCAROTENOIDS info:eu-repo/classification/Autores/CAROTENOIDS info:eu-repo/classification/cti/2 info:eu-repo/classification/cti/24 info:eu-repo/classification/cti/2415 info:eu-repo/classification/cti/241502 info:eu-repo/classification/cti/241502 ROSA YAZMIN US CAMAS Margarita de Lourdes Aguilar Espinosa Jacobo Rodríguez Campos Alba Adriana Vallejo Cardona Víctor Manuel Carballo Uicab HUGO JAVIER SERRANO POSADA Renata Lourdes Bárbara Rivera Madrid Identifying Bixa orellana L. new carotenoid cleavage dioxygenases 1 and 4 potentially involved in bixin biosynthesis |
description |
Carotene cleavage dioxygenases (CCDs) are a large family of Fe2+ dependent enzymes responsible for the production of a wide variety of apocarotenoids, such as bixin. Among the natural apocarotenoids, bixin is second in economic importance. It has a red-orange color and is produced mainly in the seeds of B. orellana. The biosynthesis of bixin aldehyde from the oxidative cleavage of lycopene at 5,6/5′,6′ bonds by a CCD is considered the first step of bixin biosynthesis. Eight BoCCD (BoCCD1-1, BoCCD1-3, BoCCD1-4, CCD4-1, BoCCD4-2, BoCCD4-3 and BoCCD4-4) genes potentially involved in the first step of B. orellana bixin biosynthesis have been identified. However, the cleavage activity upon lycopene to produce bixin aldehyde has only been demonstrated for BoCCD1-1 and BoCCD4-3. Using in vivo (Escherichia coli) and in vitro approaches, we determined that the other identified BoCCDs enzymes (BoCCD1-3, BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4) also participate in the biosynthesis of bixin aldehyde from lycopene. The LC-ESI-QTOF-MS/MS analysis showed a peak corresponding to bixin aldehyde (m/z 349.1) in pACCRT-EIB E. coli cells that express the BoCCD1 and BoCCD4 proteins, which was confirmed by in vitro enzymatic assay. Interestingly, in the in vivo assay of BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4, bixin aldehyde was oxidized to norbixin (m/z 380.2), the second product of the bixin biosynthesis pathway. In silico analysis also showed that BoCCD1 and BoCCD4 proteins encode functional dioxygenases that can use lycopene as substrate. The production of bixin aldehyde and norbixin was corroborated based on their ion fragmentation pattern, as well as by Fourier transform infrared (FTIR) spectroscopy. This work made it possible to clarify at the same time the first and second steps of the bixin biosynthesis pathway that had not been evaluated for a long time. Copyright © 2022 Us-Camas, Aguilar-Espinosa, Rodríguez-Campos, Vallejo-Cardona, Carballo-Uicab, Serrano-Posada and Rivera-Madrid. |
format |
info:eu-repo/semantics/article |
topic_facet |
info:eu-repo/classification/Autores/BoCCD1 info:eu-repo/classification/Autores/BoCCD4 info:eu-repo/classification/Autores/BIXIN ALDEHYDE info:eu-repo/classification/Autores/NORBIXIN info:eu-repo/classification/Autores/BIXIN BIOSYNTHESIS info:eu-repo/classification/Autores/BIXA ORELLANA L. info:eu-repo/classification/Autores/APOCAROTENOIDS info:eu-repo/classification/Autores/CAROTENOIDS info:eu-repo/classification/cti/2 info:eu-repo/classification/cti/24 info:eu-repo/classification/cti/2415 info:eu-repo/classification/cti/241502 info:eu-repo/classification/cti/241502 |
author |
ROSA YAZMIN US CAMAS Margarita de Lourdes Aguilar Espinosa Jacobo Rodríguez Campos Alba Adriana Vallejo Cardona Víctor Manuel Carballo Uicab HUGO JAVIER SERRANO POSADA Renata Lourdes Bárbara Rivera Madrid |
author_facet |
ROSA YAZMIN US CAMAS Margarita de Lourdes Aguilar Espinosa Jacobo Rodríguez Campos Alba Adriana Vallejo Cardona Víctor Manuel Carballo Uicab HUGO JAVIER SERRANO POSADA Renata Lourdes Bárbara Rivera Madrid |
author_sort |
ROSA YAZMIN US CAMAS |
title |
Identifying Bixa orellana L. new carotenoid cleavage dioxygenases 1 and 4 potentially involved in bixin biosynthesis |
title_short |
Identifying Bixa orellana L. new carotenoid cleavage dioxygenases 1 and 4 potentially involved in bixin biosynthesis |
title_full |
Identifying Bixa orellana L. new carotenoid cleavage dioxygenases 1 and 4 potentially involved in bixin biosynthesis |
title_fullStr |
Identifying Bixa orellana L. new carotenoid cleavage dioxygenases 1 and 4 potentially involved in bixin biosynthesis |
title_full_unstemmed |
Identifying Bixa orellana L. new carotenoid cleavage dioxygenases 1 and 4 potentially involved in bixin biosynthesis |
title_sort |
identifying bixa orellana l. new carotenoid cleavage dioxygenases 1 and 4 potentially involved in bixin biosynthesis |
url |
http://cicy.repositorioinstitucional.mx/jspui/handle/1003/2815 |
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