Cytochrome P450 Monooxygenase CYP716A141 is a Unique b-Amyrin C-16b Oxidase Involved in Triterpenoid Saponin Biosynthesis in Platycodon grandiflorus

Tamura, Keita; Teranishi, Yuga; Ueda, Seinosuke; Suzuki, Hideyuki; Kawano, Noriaki; Yoshimatsu, Kayo; Saito, Kazuki; Kawahara, Nobuo; Muranaka, Toshiya; Seki, Hikaru

doi:10.1093/pcp/pcx067

Cited by 14 publications

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“…In order to identify the most relevant of these unigenes for further analysis, we conducted BLASTx searches that compared these A. elata CYP450s to those 36 CYP450s known to be involved in triterpenoid biosynthesis. This analysis revealed that the A. elata CYP716A296 protein had 93.99% sequence identity with P. ginseng CYP716A52v2, and that CYP716A295 shared 76.94% sequence identity with P. grandiflorus CYP716A140v2, both of which are β-amyrin 28-oxidase enzymes involved in oleanolic acid production [33,39]. Moreover, the expression of CYP716A295 and CYP716A296 in different tissues were cosistent with oleanolic acid contents (Fig.…”

Section: Identification Of Candidate Cyp450s Involved In Triterpenoidmentioning

confidence: 84%

“…8; Additional file 5: Table S2). The CYP716A and CYP72A subfamilies are the primary CYP450 gene families involved in triterpenoid saponin diversification, with the CYP716A family being the largest multifunctional oxidase family involved in such triterpenoid biosynthesis [33,37,38] (Fig. 8).…”

Section: Identification Of Candidate Cyp450s Involved In Triterpenoidmentioning

confidence: 99%

“…Phylogenetic analyses revealed CYP716A295 and CYP716A296 to be grouped in the CYP716A subfamily and to be most closely related to P. grandiflorus CYP716A140v2 and P. ginseng CYP716A52v2, respectively, both of which encode β-amyrin 28-oxidase enzymes involved in oleanolic acid production [33,39]. CYP72A763 clustered in the CYP72A group and was most closely related to M. truncatula CYP72A61v2, which can transform 24-OH-β-amyrin into soyasapogenol B [34] (Fig.…”

Section: Identification Of Candidate Cyp450s Involved In Triterpenoidmentioning

confidence: 99%

“…These previous studies have suggested the CYP716 and CYP72 families to be the primary CYP450s involved in plant triterpenoid structural diversification, with their functionality having been confirmed in a wide range of plant types. Members of the CYP716 family have been found to often exhibit pentacyclic triterpenoid C-28 oxidation activity, as in the case of the Panax ginseng enzyme CYP716A52v2 [32] and the Platycodon grandiflorus enzymes CYP716A140v2 and CYP716A141 [33]. Members of the CYP72A subfamily, in contrast, primarily exhibit oleanolic acid C-23 or C-2β hydroxylase activity.…”

Section: Introductionmentioning

confidence: 99%

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Identification and analysis of CYP450 supergene family members from the transcriptome of Aralia elata (Miq.) Seem reveal candidate genes for triterpenoid saponins biosynthesis

Cheng

Liu

Tong

et al. 2020

Preprint

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BackgroundMembers of the cytochrome P450 (CYP450) gene superfamily have been shown to play essential roles in regulating secondary metabolites biosynthesis. However, the systematic identification and bioinformatics analysis of CYP450s have not been reported in Aralia elata (Miq.) Seem, a highly valued medicinal plant. ResultsIn the present study we conducted the RNA-sequencing (RNA-seq) analysis of the leaves, stems, and roots of A. elata, yielding 66,713 total unigenes. Following the annotation and classification of these unigenes, we were able to identify two pathways and 19 putative genes associated with the synthesis of triterpenoid saponins in these plants, with qRT-PCR subsequently being used to validate these gene expression patterns. Scanning with the CYP450 model from Pfam resulted in the identification of 111 full-length and 143 partial-length CYP450s, with the full-length CYP450s being further clustered into 7 clans and 36 families. Through phylogenetic and conserved motif analyses, we were further able to group these CYP450 proteins into two primary branches: A-type (53%) and non-A type (47%). We further conducted representative protein sequence alignment for these CYP450 family members, with secondary elements being assigned in light of the recently published Arabidopsis CYP90B1 structure. Using the available sequence information, we further identified predicted substrate recognition sites (SRSs) and substrate binding sites within these putative proteins.We further assessed the expression patterns of these 111 CYP450 genes across A. elata tissues, with 12 members of this gene family being selected at random for qRT-PCR validation. From these data, we identified CYP716A295 and CYP716A296 as the candidate genes most likely to be associated with oleanolic acid synthesis, while CYP72A763 was identified as being the most likely to play a role in hederagenin biosynthesis. Finally, we assessed the subcellular localization of these CYP450 proteins within Arabidopsis protoplasts, highlighting the fact that they localize to the endoplasmic reticulum.ConclusionsThis study presents a systematic analysis of the CYP450 gene family in A. elata and provided a foundation for further functional characterization of CYP450 genes.

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Section: Identification Of Candidate Cyp450s Involved In Triterpenoidmentioning

confidence: 84%

Section: Identification Of Candidate Cyp450s Involved In Triterpenoidmentioning

confidence: 99%

Section: Identification Of Candidate Cyp450s Involved In Triterpenoidmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Identification and analysis of CYP450 supergene family members from the transcriptome of Aralia elata (Miq.) Seem reveal candidate genes for triterpenoid saponins biosynthesis

Cheng

Liu

Tong

et al. 2020

Preprint

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“…Advanced sequencing technology helps us to discover new genes involved in secondary metabolite formation and to infer the biosynthesis pathway for medicinal plants. De novo transcriptome studies have been successfully used to infer the saponin biosynthesis pathway in plants such as Panax japonica , Anemone flaccida, Ilex asprella , and Platycodon grandiflorus [ 26 – 29 ].…”

Section: Discussionmentioning

confidence: 99%

Identification of potential genes involved in triterpenoid saponins biosynthesis in Gleditsia sinensis by transcriptome and metabolome analyses

et al. 2018

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Gleditsia sinensis is widely used as a medicinal plant in Asia, especially in China. Triterpenes, alkaloids, and sterols were isolated from Gleditsia species. Among them, triterpenoid saponins are very important metabolites owing to their various pharmacological activities. However, the triterpenoid saponin biosynthesis pathway has not been well characterized. In the present study, we performed de novo transcriptome assembly for 14.3 Gbps of clean reads sequenced from nine tissues of G. sinensis . The results showed that 81,511 unique transcripts (unitranscripts) (47,855 unigenes) were constructed, of which 31,717 unigenes were annotated with Gene Ontology and EC numbers by Blast2GO against the NCBI-nr protein database. We also analyzed the metabolite contents in the same nine tissues by LS–MS/MS, and saponins including gleditsioside I were found in fruit at higher levels. Many of the genes with tissue-specific expression in fruit are involved in the flavonoid biosynthesis pathway, and many of those have UDP-glucosyltransferase (UGT) activity. We constructed a saponin biosynthesis pathway and identified two key enzyme families in the triterpenoid saponin biosynthesis pathway, cytochrome P450 and UDP-glucosyltransferase, that are encoded by 37 unigenes and 77 unigenes, respectively. CYP72A, CYP716A, and CYP88D, which are known as key enzymes for saponin biosynthesis, were also identified among the P450s. Our results provide insight into the secondary metabolite biosynthesis and serve as important resources for future research and cultivation of G. sinensis . Electronic supplementary material The online version of this article (10.1007/s11418-018-1270-2) contains supplementary material, which is available to authorized users.

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Production of plant bioactive triterpenoid saponins: from metabolites to genes and back

2020

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Cytochrome P450 Monooxygenase CYP716A141 is a Unique b-Amyrin C-16b Oxidase Involved in Triterpenoid Saponin Biosynthesis in Platycodon grandiflorus

Cited by 14 publications

References 0 publications

Identification and analysis of CYP450 supergene family members from the transcriptome of Aralia elata (Miq.) Seem reveal candidate genes for triterpenoid saponins biosynthesis

Identification and analysis of CYP450 supergene family members from the transcriptome of Aralia elata (Miq.) Seem reveal candidate genes for triterpenoid saponins biosynthesis

Identification of potential genes involved in triterpenoid saponins biosynthesis in Gleditsia sinensis by transcriptome and metabolome analyses

Production of plant bioactive triterpenoid saponins: from metabolites to genes and back

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