The Functional Characteristics and Soluble Expression of Saffron <i>Cs</i>CCD2
Crocins are important natural products predominantly obtained from the stigma of saffron, and that can be utilized as a medicinal compound, spice, and colorant with significant promise in the pharmaceutical, food, and cosmetic industries. Carotenoid cleavage dioxygenase 2 (<i>Cs</i>CCD2)...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-10-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/20/15090 |
| Summary: | Crocins are important natural products predominantly obtained from the stigma of saffron, and that can be utilized as a medicinal compound, spice, and colorant with significant promise in the pharmaceutical, food, and cosmetic industries. Carotenoid cleavage dioxygenase 2 (<i>Cs</i>CCD2) is a crucial limiting enzyme that has been reported to be responsible for the cleavage of zeaxanthin in the crocin biosynthetic pathway. However, the catalytic activity of <i>Cs</i>CCD2 on β-carotene/lycopene remains elusive, and the soluble expression of <i>Cs</i>CCD2 remains a big challenge. In this study, we reported the functional characteristics of <i>Cs</i>CCD2, that can catalyze not only zeaxanthin cleavage but also β-carotene and lycopene cleavage. The molecular basis of the divergent functionality of <i>Cs</i>CCD2 was elucidated using bioinformatic analysis and truncation studies. The protein expression optimization results demonstrated that the use of a maltose-binding protein (MBP) tag and the optimization of the induction conditions resulted in the production of more soluble protein. Correspondingly, the catalytic efficiency of soluble <i>Cs</i>CCD2 was higher than that of the insoluble one, and the results further validated its functional verification. This study not only broadened the substrate profile of <i>Cs</i>CCD2, but also achieved the soluble expression of <i>Cs</i>CCD2. It provides a firm platform for <i>Cs</i>CCD2 crystal structure resolution and facilitates the synthesis of crocetin and crocins. |
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| ISSN: | 1661-6596 1422-0067 |