Effects of high light exposure and heterologous expression of β-carotene ketolase on the metabolism of carotenoids in Chlamydomonas reinhardtii
IntroductionStress from high light exposure and overexpression of β-carotene ketolase can have significant effects on the synthesis of carotenoids in Chlamydomonas reinhardtii. As a promising platform for carotenoid production, C. reinhardtii needs further research and technological innovation to ad...
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Frontiers Media S.A.
2025-03-01
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| author | Rui Mei Haihong Yang Chunli Guo Zeyu Hong Zhangli Hu Zhangli Hu Yan Wu Danqiong Huang Chaogang Wang Chaogang Wang |
| author_facet | Rui Mei Haihong Yang Chunli Guo Zeyu Hong Zhangli Hu Zhangli Hu Yan Wu Danqiong Huang Chaogang Wang Chaogang Wang |
| author_sort | Rui Mei |
| collection | DOAJ |
| description | IntroductionStress from high light exposure and overexpression of β-carotene ketolase can have significant effects on the synthesis of carotenoids in Chlamydomonas reinhardtii. As a promising platform for carotenoid production, C. reinhardtii needs further research and technological innovation to address challenges, such as environmental interference, exogenous gene expression, and metabolic regulation, to achieve efficient and sustainable production of carotenoids.MethodsAppropriate β-carotene ketolase were selected from different organisms and subjected for codon optimization based on the preferences of the nuclear genome of C. reinhardtii. After designation, including intron insertion and chloroplast transit peptide, expression vectors were constructed and used for nuclear transformation of C. reinhardtii CC849 by bead milling method. Subsequently, DNA-PCR and RT-PCR were used to identify positive transformants grown with antibiotic stress, LC-MS/MS and metabolic analysis were performed to evaluate the products of transformants.ResultsIn this study, carotenoid metabolism regulation in C. reinhardtii was investigated in a time-dependent manner through high light exposure and heterologous expression of β-carotene ketolase. The results suggested that the stress from high light exposure (500 μmol/m2/s) negatively regulated the accumulation of β-carotene; positively induced the accumulation of zeaxanthin, echinenone, and canthaxanthin; and continuously promoted accumulation of zeaxanthin and canthaxanthin in C. reinhardtii. Metabolomics analysis suggested that high light exposure stress promoted biosynthesis of carotenoids, improved the intermediates associated with the astaxanthin synthesis pathway, and promoted conversion of β-carotene to downstream substances. Several strategies were implemented to improve canthaxanthin production in C. reinhardtii to achieve overexpression of β-carotene ketolase genes from different sources, including strong promoters, insertion introns, and chloroplast conduction peptides. It was found that β-carotene, echinenone, and canthaxanthin were all significantly increased in the transformed C. reinhardtii overexpressing β-carotene ketolase. Among these, the highest canthaxanthin content was found in pH124-CrtO, which was seven times that observed in the wild type. Moreover, the metabolomics analysis of carotenoids showed promotion of the abscisic acid and astaxanthin pathways in the transformed C. reinhardtii.DiscussionThe results of this study provide a new scheme for manipulating the metabolism of carotenoids and promoting the synthesis of high-value carotenoids in C. reinhardtii. |
| format | Article |
| id | doaj-art-33d6ffda65714e38b831d65bd168f48d |
| institution | DOAJ |
| issn | 2296-4185 |
| language | English |
| publishDate | 2025-03-01 |
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| spelling | doaj-art-33d6ffda65714e38b831d65bd168f48d2025-08-20T02:52:55ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852025-03-011310.3389/fbioe.2025.15336611533661Effects of high light exposure and heterologous expression of β-carotene ketolase on the metabolism of carotenoids in Chlamydomonas reinhardtiiRui Mei0Haihong Yang1Chunli Guo2Zeyu Hong3Zhangli Hu4Zhangli Hu5Yan Wu6Danqiong Huang7Chaogang Wang8Chaogang Wang9Shenzhen Key Laboratory of Marine Bioresource and Eco-Environmental Science, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, ChinaShenzhen Key Laboratory of Marine Bioresource and Eco-Environmental Science, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, ChinaShenzhen Key Laboratory of Marine Bioresource and Eco-Environmental Science, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, ChinaShenzhen Key Laboratory of Marine Bioresource and Eco-Environmental Science, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, ChinaInstrumental Analysis Center, Shenzhen University, Shenzhen, ChinaShenzhen Key Laboratory of Marine Bioresource and Eco-Environmental Science, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, ChinaShenzhen Key Laboratory of Marine Bioresource and Eco-Environmental Science, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, ChinaGuangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, ChinaIntroductionStress from high light exposure and overexpression of β-carotene ketolase can have significant effects on the synthesis of carotenoids in Chlamydomonas reinhardtii. As a promising platform for carotenoid production, C. reinhardtii needs further research and technological innovation to address challenges, such as environmental interference, exogenous gene expression, and metabolic regulation, to achieve efficient and sustainable production of carotenoids.MethodsAppropriate β-carotene ketolase were selected from different organisms and subjected for codon optimization based on the preferences of the nuclear genome of C. reinhardtii. After designation, including intron insertion and chloroplast transit peptide, expression vectors were constructed and used for nuclear transformation of C. reinhardtii CC849 by bead milling method. Subsequently, DNA-PCR and RT-PCR were used to identify positive transformants grown with antibiotic stress, LC-MS/MS and metabolic analysis were performed to evaluate the products of transformants.ResultsIn this study, carotenoid metabolism regulation in C. reinhardtii was investigated in a time-dependent manner through high light exposure and heterologous expression of β-carotene ketolase. The results suggested that the stress from high light exposure (500 μmol/m2/s) negatively regulated the accumulation of β-carotene; positively induced the accumulation of zeaxanthin, echinenone, and canthaxanthin; and continuously promoted accumulation of zeaxanthin and canthaxanthin in C. reinhardtii. Metabolomics analysis suggested that high light exposure stress promoted biosynthesis of carotenoids, improved the intermediates associated with the astaxanthin synthesis pathway, and promoted conversion of β-carotene to downstream substances. Several strategies were implemented to improve canthaxanthin production in C. reinhardtii to achieve overexpression of β-carotene ketolase genes from different sources, including strong promoters, insertion introns, and chloroplast conduction peptides. It was found that β-carotene, echinenone, and canthaxanthin were all significantly increased in the transformed C. reinhardtii overexpressing β-carotene ketolase. Among these, the highest canthaxanthin content was found in pH124-CrtO, which was seven times that observed in the wild type. Moreover, the metabolomics analysis of carotenoids showed promotion of the abscisic acid and astaxanthin pathways in the transformed C. reinhardtii.DiscussionThe results of this study provide a new scheme for manipulating the metabolism of carotenoids and promoting the synthesis of high-value carotenoids in C. reinhardtii.https://www.frontiersin.org/articles/10.3389/fbioe.2025.1533661/fullcarotenoidscanthaxanthinChlamydomonas reinhardtiiheterologous expressionmetabolic analysis |
| spellingShingle | Rui Mei Haihong Yang Chunli Guo Zeyu Hong Zhangli Hu Zhangli Hu Yan Wu Danqiong Huang Chaogang Wang Chaogang Wang Effects of high light exposure and heterologous expression of β-carotene ketolase on the metabolism of carotenoids in Chlamydomonas reinhardtii Frontiers in Bioengineering and Biotechnology carotenoids canthaxanthin Chlamydomonas reinhardtii heterologous expression metabolic analysis |
| title | Effects of high light exposure and heterologous expression of β-carotene ketolase on the metabolism of carotenoids in Chlamydomonas reinhardtii |
| title_full | Effects of high light exposure and heterologous expression of β-carotene ketolase on the metabolism of carotenoids in Chlamydomonas reinhardtii |
| title_fullStr | Effects of high light exposure and heterologous expression of β-carotene ketolase on the metabolism of carotenoids in Chlamydomonas reinhardtii |
| title_full_unstemmed | Effects of high light exposure and heterologous expression of β-carotene ketolase on the metabolism of carotenoids in Chlamydomonas reinhardtii |
| title_short | Effects of high light exposure and heterologous expression of β-carotene ketolase on the metabolism of carotenoids in Chlamydomonas reinhardtii |
| title_sort | effects of high light exposure and heterologous expression of β carotene ketolase on the metabolism of carotenoids in chlamydomonas reinhardtii |
| topic | carotenoids canthaxanthin Chlamydomonas reinhardtii heterologous expression metabolic analysis |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2025.1533661/full |
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