Cargando…

Functional Characterization of Lycopene β- and ε-Cyclases from a Lutein-Enriched Green Microalga Chlorella sorokiniana FZU60

Lutein is a high-value carotenoid with many human health benefits. Lycopene β- and ε-cyclases (LCYB and LCYE, respectively) catalyze the cyclization of lycopene into distinct downstream branches, one of which is the lutein biosynthesis pathway, via α-carotene. Hence, LCYB and LCYE are key enzymes in...

Descripción completa

Detalles Bibliográficos
Autores principales: Fang, Hong, Liu, Junjie, Ma, Ruijuan, Zou, Yiping, Ho, Shih-Hsin, Chen, Jianfeng, Xie, Youping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10381880/
https://www.ncbi.nlm.nih.gov/pubmed/37504949
http://dx.doi.org/10.3390/md21070418
_version_ 1785080553585770496
author Fang, Hong
Liu, Junjie
Ma, Ruijuan
Zou, Yiping
Ho, Shih-Hsin
Chen, Jianfeng
Xie, Youping
author_facet Fang, Hong
Liu, Junjie
Ma, Ruijuan
Zou, Yiping
Ho, Shih-Hsin
Chen, Jianfeng
Xie, Youping
author_sort Fang, Hong
collection PubMed
description Lutein is a high-value carotenoid with many human health benefits. Lycopene β- and ε-cyclases (LCYB and LCYE, respectively) catalyze the cyclization of lycopene into distinct downstream branches, one of which is the lutein biosynthesis pathway, via α-carotene. Hence, LCYB and LCYE are key enzymes in lutein biosynthesis. In this study, the coding genes of two lycopene cyclases (CsLCYB and CsLCYE) of a lutein-enriched marine green microalga, Chlorella sorokiniana FZU60, were isolated and identified. A sequence analysis and computational modeling of CsLCYB and CsLCYE were performed using bioinformatics to identify the key structural domains. Further, a phylogenetic analysis revealed that CsLCYB and CsLCYE were homogeneous to the proteins of other green microalgae. Subcellular localization tests in Nicotiana benthamiana showed that CsLCYB and CsLCYE localized in chloroplasts. A pigment complementation assay in Escherichia coli revealed that CsLCYB could efficiently β-cyclize both ends of lycopene to produce β-carotene. On the other hand, CsLCYE possessed a strong ε-monocyclase activity for the production of δ-carotene and a weak ε-bicyclic activity for the production of ε-carotene. In addition, CsLCYE was able to catalyze lycopene into β-monocyclic γ-carotene and ultimately produced α-carotene with a β-ring and an ε-ring via γ-carotene or δ-carotene. Moreover, the co-expression of CsLCYB and CsLCYE in E. coli revealed that α-carotene was a major product, which might lead to the production of a high level of lutein in C. sorokiniana FZU60. The findings provide a theoretical foundation for performing metabolic engineering to improve lutein biosynthesis and accumulation in C. sorokiniana FZU60.
format Online
Article
Text
id pubmed-10381880
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-103818802023-07-29 Functional Characterization of Lycopene β- and ε-Cyclases from a Lutein-Enriched Green Microalga Chlorella sorokiniana FZU60 Fang, Hong Liu, Junjie Ma, Ruijuan Zou, Yiping Ho, Shih-Hsin Chen, Jianfeng Xie, Youping Mar Drugs Article Lutein is a high-value carotenoid with many human health benefits. Lycopene β- and ε-cyclases (LCYB and LCYE, respectively) catalyze the cyclization of lycopene into distinct downstream branches, one of which is the lutein biosynthesis pathway, via α-carotene. Hence, LCYB and LCYE are key enzymes in lutein biosynthesis. In this study, the coding genes of two lycopene cyclases (CsLCYB and CsLCYE) of a lutein-enriched marine green microalga, Chlorella sorokiniana FZU60, were isolated and identified. A sequence analysis and computational modeling of CsLCYB and CsLCYE were performed using bioinformatics to identify the key structural domains. Further, a phylogenetic analysis revealed that CsLCYB and CsLCYE were homogeneous to the proteins of other green microalgae. Subcellular localization tests in Nicotiana benthamiana showed that CsLCYB and CsLCYE localized in chloroplasts. A pigment complementation assay in Escherichia coli revealed that CsLCYB could efficiently β-cyclize both ends of lycopene to produce β-carotene. On the other hand, CsLCYE possessed a strong ε-monocyclase activity for the production of δ-carotene and a weak ε-bicyclic activity for the production of ε-carotene. In addition, CsLCYE was able to catalyze lycopene into β-monocyclic γ-carotene and ultimately produced α-carotene with a β-ring and an ε-ring via γ-carotene or δ-carotene. Moreover, the co-expression of CsLCYB and CsLCYE in E. coli revealed that α-carotene was a major product, which might lead to the production of a high level of lutein in C. sorokiniana FZU60. The findings provide a theoretical foundation for performing metabolic engineering to improve lutein biosynthesis and accumulation in C. sorokiniana FZU60. MDPI 2023-07-23 /pmc/articles/PMC10381880/ /pubmed/37504949 http://dx.doi.org/10.3390/md21070418 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Fang, Hong
Liu, Junjie
Ma, Ruijuan
Zou, Yiping
Ho, Shih-Hsin
Chen, Jianfeng
Xie, Youping
Functional Characterization of Lycopene β- and ε-Cyclases from a Lutein-Enriched Green Microalga Chlorella sorokiniana FZU60
title Functional Characterization of Lycopene β- and ε-Cyclases from a Lutein-Enriched Green Microalga Chlorella sorokiniana FZU60
title_full Functional Characterization of Lycopene β- and ε-Cyclases from a Lutein-Enriched Green Microalga Chlorella sorokiniana FZU60
title_fullStr Functional Characterization of Lycopene β- and ε-Cyclases from a Lutein-Enriched Green Microalga Chlorella sorokiniana FZU60
title_full_unstemmed Functional Characterization of Lycopene β- and ε-Cyclases from a Lutein-Enriched Green Microalga Chlorella sorokiniana FZU60
title_short Functional Characterization of Lycopene β- and ε-Cyclases from a Lutein-Enriched Green Microalga Chlorella sorokiniana FZU60
title_sort functional characterization of lycopene β- and ε-cyclases from a lutein-enriched green microalga chlorella sorokiniana fzu60
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10381880/
https://www.ncbi.nlm.nih.gov/pubmed/37504949
http://dx.doi.org/10.3390/md21070418
work_keys_str_mv AT fanghong functionalcharacterizationoflycopenebandecyclasesfromaluteinenrichedgreenmicroalgachlorellasorokinianafzu60
AT liujunjie functionalcharacterizationoflycopenebandecyclasesfromaluteinenrichedgreenmicroalgachlorellasorokinianafzu60
AT maruijuan functionalcharacterizationoflycopenebandecyclasesfromaluteinenrichedgreenmicroalgachlorellasorokinianafzu60
AT zouyiping functionalcharacterizationoflycopenebandecyclasesfromaluteinenrichedgreenmicroalgachlorellasorokinianafzu60
AT hoshihhsin functionalcharacterizationoflycopenebandecyclasesfromaluteinenrichedgreenmicroalgachlorellasorokinianafzu60
AT chenjianfeng functionalcharacterizationoflycopenebandecyclasesfromaluteinenrichedgreenmicroalgachlorellasorokinianafzu60
AT xieyouping functionalcharacterizationoflycopenebandecyclasesfromaluteinenrichedgreenmicroalgachlorellasorokinianafzu60