A DUF4281 domain-containing protein (homologue of ABA4) of Phaeodactylum tricornutum regulates the biosynthesis of fucoxanthin

Shen, Xiaotong; Pan, Kehou; Zhang, Lin; Zhu, Bo; Li, Yun; Han, Jichang

doi:10.1016/j.algal.2022.102728

Cited by 8 publications

(5 citation statements)

References 63 publications

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“…Specifically, P. tricornutum comprises Fx (10.67 mg/g), diadinoxanthin (3.26 mg/g), diatoxanthin (0.20 mg/g), violaxanthin (0.05 mg/g), and zeaxanthin (0.04 mg/g) as measured through HPLC analysis. Neoxanthin was not detected [ 29 ].…”

Section: Discussionmentioning

confidence: 99%

“…However, in vitro neoxanthin synthase activity was not observed PtABA4 when was expressed in E. coli . It has been suggested that PtABA4 forms multi-enzyme complexes with other membrane proteins, indicating its exclusive involvement in carotenoid formation instead of neoxanthin [ 29 ]. Therefore, to enhance Fx biosynthesis, in addition to the overexpression of PtVDL1, the activity of downstream enzymes must be increased.…”

Section: Discussionmentioning

confidence: 99%

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Overexpression of PtVDL1 in Phaeodactylum tricornutum Increases Fucoxanthin Content under Red Light

Seo,

Chang,

Choi

et al. 2023

J. Microbiol. Biotechnol.

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Phaeodactylum tricornutum is a model diatom with significant biotechnological applications, including enhancing biomass, biofuel, and carotenoid production. Specifically, owing to the capacity of this organism to serve as a valuable source of essential raw materials for pharmaceuticals and nutraceuticals, ongoing research is actively focused on enhancing its productivity. One of the genes involved in various stages of fucoxanthin (Fx) biosynthesis, violaxanthin de-epoxidase like 1 (VDL1), has recently been identified. To validate the intracellular function of this gene and boost Fx production through overexpression, we established and examined three transgenic P. tricornutum lines characterized by elevated P. tricortunum VDL1 ( PtVDL1) expression and evaluate their cell growth and Fx productivity. These transgenic lines exhibited substantially increased PtVDL1 mRNA and protein levels compared to the wild type (WT). Notably, the enzyme substrate violaxanthin was entirely depleted and could not be detected in the transformants, whereas it remained at constant levels in the WT. Interestingly, under standard white light conditions, Fx productivity in the transformants remained unchanged; however, but after 48 h of exposure to red light, it increased by up to 15%. These results indicate that PtVDL1-overexpressing P. tricornutum has industrial potential, particularly for enhancing Fx production under red light conditions.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Overexpression of PtVDL1 in Phaeodactylum tricornutum Increases Fucoxanthin Content under Red Light

Seo,

Chang,

Choi

et al. 2023

J. Microbiol. Biotechnol.

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“…Although many genes involved in carotenoid biosynthesis have been tentatively identified and summarized, there are still few reports about the putative genes involved in the conversion of violaxanthin to neoxanthin. Shen et al identified a gene named PtABA4 that has a potential function relevant to violaxanthin conversion (Shen et al, 2022). However, the change in neoxanthin accumulation was not observed with the expression of PtABA4 .…”

Section: Discussionmentioning

confidence: 99%

“…The copyright holder for this preprint this version posted October 18, 2022. ; https://doi.org/10.1101/2022.10.18.512692 doi: bioRxiv preprint function relevant to violaxanthin conversion (Shen et al, 2022). However, the change in neoxanthin accumulation was not observed with the expression of PtABA4.…”

Section: Nxs Regulates the Accumulations Of Xanthophyllsmentioning

confidence: 96%

Isolation and characterization of a neoxanthin synthase gene functioning in fucoxanthin biosynthesis ofPhaeodactyum tricornutum

Kang

Zhang

Han

et al. 2022

Preprint

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Golden-brown xanthophyll fucoxanthin in marine organisms, especially in diatoms, has attracted widespread attention because of its diverse biological activities. However, the biosynthetic pathway of fucoxanthin remains unclear in diatoms. Fucoxanthin may derive from either neoxanthin or diadinoxanthin pathway. However, the key point is whether neoxanthin and its synthesizing genes exist or not. In this study, we successfully identified a few xanthophylls in trace amounts in the concentrated fraction of carotenoids of diatom Phaeodactylum tricornutum cultured at different light intensities with the co-chromatography method, and cloned the neoxanthin synthase (NXS) gene which was not annotated in diatom genome. The NXS knockdown and knockout experiment show a positive correlation in the accumulation of neoxanthin and zeaxanthin while a negative correlation in violaxanthin and fucoxanthin with the expression of NXS. In vitro assay evidenced that neoxanthin is the precursor for fucoxanthin biosynthesis, indicating that other molecules intermediate the conversion between violaxanthin and fucoxanthin. Overall, we cloned a novel gene functioning in neoxanthin biosynthesis, which should aid to clarifying the fucoxanthin biosynthetic pathway in diatom.

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“…During the batch cultivation period, OD 750 of microalgae cultures was measured every 2 days by Thermo Fisher Scientific Microplater Reader (Varioskan LUX, Finland) to represent the cell concentration ( Shen et al, 2022 ; Zhang et al, 2022 ). At the final cultivation day (Day 10), 10 mL of C. roscoffensis cultures collected by Whatman GF/F filters (0.70 µm) were rinsed twice with Milli-Q water and dried to constant weight (60°C for 48 h) to determine the biomass.…”

Section: Methodsmentioning

confidence: 99%

Screening of antibiotics to obtain axenic cell cultures of a marine microalga Chrysotila roscoffensis

Liu

Sun

Zhang

et al. 2023

Front. Bioeng. Biotechnol.

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Due to high growth rate, outstanding abiotic stress tolerance, and rich value-added substances, Chrysotila roscoffensis, belonging to the phylum of Haptophyta, can be considered as a versatile resource for industrial exploitation of bioactive compounds. However, the application potential of C. roscoffensis has drawn attention until just recently, and the understanding related to the biological properties of this species is still scarce. For example, the sensitivities of C. roscoffensis to antibiotics, which is essential for the verification of heterotrophic capacity and the establishment of efficient genetic manipulation system is still unavailable. Aiming to provide fundamental information for future exploitation, the sensitivities of C. roscoffensis to nine types of antibiotics were tested in this study. The results demonstrated that C. roscoffensis exhibited relatively high resistances to ampicillin, kanamycin, streptomycin, gentamicin, and geneticin, while was sensitive to bleomycin, hygromycin B, paromomycin, and chloramphenicol. Using the former five types of antibiotics, a bacteria removal strategy was established tentatively. Finally, the axenicity of treated C. roscoffensis was confirmed based on a multi-strategy method including solid plate, 16S rDNA amplification, and nuclear acid staining. This report can provide valuable information for the development of optimal selection markers, which are meaningful for more extensive transgenic studies in C. roscoffensis. Moreover, our study also paves the way for the establishment of heterotrophic/mixotrophic cultivation modes of C. roscoffensis.

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A DUF4281 domain-containing protein (homologue of ABA4) of Phaeodactylum tricornutum regulates the biosynthesis of fucoxanthin

Cited by 8 publications

References 63 publications

Overexpression of PtVDL1 in Phaeodactylum tricornutum Increases Fucoxanthin Content under Red Light

Overexpression of PtVDL1 in Phaeodactylum tricornutum Increases Fucoxanthin Content under Red Light

Isolation and characterization of a neoxanthin synthase gene functioning in fucoxanthin biosynthesis ofPhaeodactyum tricornutum

Screening of antibiotics to obtain axenic cell cultures of a marine microalga Chrysotila roscoffensis

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