Characteristics of NtCCD1-3 from tobacco, and protein engineering of the CCD1 to enhance β-ionone production in yeast
Biosynthesis of β-ionone by microbial cell factories has become a promising way to obtain natural β-ionone. The catalytic activity of carotenoid cleavage dioxygenase 1 (CCD1) in cleavage of β-carotene to β-ionone severely limits its biosynthesis. In this study, NtCCD1-3 from Nicotiana tabacum with h...
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| Format: | Article |
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Frontiers Media S.A.
2022-09-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2022.1011297/full |
| _version_ | 1818058153861840896 |
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| author | Xiaowei Gong Xiaowei Gong Fan Li Yupeng Liang Xiulin Han Mengliang Wen |
| author_facet | Xiaowei Gong Xiaowei Gong Fan Li Yupeng Liang Xiulin Han Mengliang Wen |
| author_sort | Xiaowei Gong |
| collection | DOAJ |
| description | Biosynthesis of β-ionone by microbial cell factories has become a promising way to obtain natural β-ionone. The catalytic activity of carotenoid cleavage dioxygenase 1 (CCD1) in cleavage of β-carotene to β-ionone severely limits its biosynthesis. In this study, NtCCD1-3 from Nicotiana tabacum with high ability to cleave β-carotene was screened. Multiple strategies for improving the β-ionone yield in Saccharomyces cerevisiae were performed. The results showed that NtCCD1-3 could cleave a variety of caroteniods at the 9,10 (9′,10′) double bonds and lycopene at the 5,6 (5′,6′) positions. The insertion site delta for NtCCD1-3 gene was more suitable for enhancing the yield of β-ionone, showing 19.1-fold increase compared with the rox1 site. More importantly, mutant K38A of NtCCD1-3 in membrane-bonding domains could greatly promote β-ionone production by more than 3-fold. We also found that overexpression of the NADH kinase Pos5 could improve β-ionone yield up to 1.5 times. These results may provide valuable references for biosynthesis of β-ionone. |
| first_indexed | 2024-12-10T12:56:07Z |
| format | Article |
| id | doaj.art-d90e737f0a424dbdbcfddb36cff37311 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-10T12:56:07Z |
| publishDate | 2022-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-d90e737f0a424dbdbcfddb36cff373112022-12-22T01:48:06ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-09-011310.3389/fmicb.2022.10112971011297Characteristics of NtCCD1-3 from tobacco, and protein engineering of the CCD1 to enhance β-ionone production in yeastXiaowei Gong0Xiaowei Gong1Fan Li2Yupeng Liang3Xiulin Han4Mengliang Wen5National Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Key Laboratory of Microbial Diversity in Southwest China, Ministry of Education, School of Life Sciences, Yunnan Institute of Microbiology, Yunnan University, Kunming, ChinaR&D Center, China Tobacco Yunnan Industrial Co., Ltd., Kunming, ChinaNational Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Key Laboratory of Microbial Diversity in Southwest China, Ministry of Education, School of Life Sciences, Yunnan Institute of Microbiology, Yunnan University, Kunming, ChinaNational Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Key Laboratory of Microbial Diversity in Southwest China, Ministry of Education, School of Life Sciences, Yunnan Institute of Microbiology, Yunnan University, Kunming, ChinaNational Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Key Laboratory of Microbial Diversity in Southwest China, Ministry of Education, School of Life Sciences, Yunnan Institute of Microbiology, Yunnan University, Kunming, ChinaNational Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Key Laboratory of Microbial Diversity in Southwest China, Ministry of Education, School of Life Sciences, Yunnan Institute of Microbiology, Yunnan University, Kunming, ChinaBiosynthesis of β-ionone by microbial cell factories has become a promising way to obtain natural β-ionone. The catalytic activity of carotenoid cleavage dioxygenase 1 (CCD1) in cleavage of β-carotene to β-ionone severely limits its biosynthesis. In this study, NtCCD1-3 from Nicotiana tabacum with high ability to cleave β-carotene was screened. Multiple strategies for improving the β-ionone yield in Saccharomyces cerevisiae were performed. The results showed that NtCCD1-3 could cleave a variety of caroteniods at the 9,10 (9′,10′) double bonds and lycopene at the 5,6 (5′,6′) positions. The insertion site delta for NtCCD1-3 gene was more suitable for enhancing the yield of β-ionone, showing 19.1-fold increase compared with the rox1 site. More importantly, mutant K38A of NtCCD1-3 in membrane-bonding domains could greatly promote β-ionone production by more than 3-fold. We also found that overexpression of the NADH kinase Pos5 could improve β-ionone yield up to 1.5 times. These results may provide valuable references for biosynthesis of β-ionone.https://www.frontiersin.org/articles/10.3389/fmicb.2022.1011297/fullcarotenoid cleavage dioxygenase 1Nicotiana tabacumβ-iononeprotein engineeringSaccharomyces cerevisiae |
| spellingShingle | Xiaowei Gong Xiaowei Gong Fan Li Yupeng Liang Xiulin Han Mengliang Wen Characteristics of NtCCD1-3 from tobacco, and protein engineering of the CCD1 to enhance β-ionone production in yeast Frontiers in Microbiology carotenoid cleavage dioxygenase 1 Nicotiana tabacum β-ionone protein engineering Saccharomyces cerevisiae |
| title | Characteristics of NtCCD1-3 from tobacco, and protein engineering of the CCD1 to enhance β-ionone production in yeast |
| title_full | Characteristics of NtCCD1-3 from tobacco, and protein engineering of the CCD1 to enhance β-ionone production in yeast |
| title_fullStr | Characteristics of NtCCD1-3 from tobacco, and protein engineering of the CCD1 to enhance β-ionone production in yeast |
| title_full_unstemmed | Characteristics of NtCCD1-3 from tobacco, and protein engineering of the CCD1 to enhance β-ionone production in yeast |
| title_short | Characteristics of NtCCD1-3 from tobacco, and protein engineering of the CCD1 to enhance β-ionone production in yeast |
| title_sort | characteristics of ntccd1 3 from tobacco and protein engineering of the ccd1 to enhance β ionone production in yeast |
| topic | carotenoid cleavage dioxygenase 1 Nicotiana tabacum β-ionone protein engineering Saccharomyces cerevisiae |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2022.1011297/full |
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