scholarly journals Transcriptomic Insight into Terpenoid Biosynthesis and Functional Characterization of Three Diterpene Synthases in Scutellaria barbata

Molecules ◽  
2018 ◽  
Vol 23 (11) ◽  
pp. 2952 ◽  
Author(s):  
Huabei Zhang ◽  
Baolong Jin ◽  
Junling Bu ◽  
Juan Guo ◽  
Tong Chen ◽  
...  
Keyword(s):  
Average Length ◽  
Functional Characterization ◽  
Terpene Synthase ◽  
Terpenoid Biosynthesis ◽  
Vitro Assay ◽  
Scutellaria Barbata ◽  
Aerial Parts ◽  
Diterpene Synthase ◽  
Clerodane Diterpenoids

Scutellaria barbata (Lamiaceae) is an important medicinal herb widely used in China, Korea, India, and other Asian countries. Neo-clerodane diterpenoids are the largest known group of Scutellaria diterpenoids and show promising cytotoxic activity against several cancer cell lines. Here, Illumina-based deep transcriptome analysis of flowers, the aerial parts (leaf and stem), and roots of S. barbata was used to explore terpenoid-related genes. In total, 121,958,564 clean RNA-sequence reads were assembled into 88,980 transcripts, with an average length of 1370 nt and N50 length of 2144 nt, indicating high assembly quality. We identified nearly all known terpenoid-related genes (33 genes) involved in biosynthesis of the terpenoid backbone and 14 terpene synthase genes which generate skeletons for different terpenoids. Three full length diterpene synthase genes were functionally identified using an in vitro assay. SbTPS8 and SbTPS9 were identified as normal-CPP and ent-CPP synthase, respectively. SbTPS12 reacts with SbTPS8 to produce miltiradiene. Furthermore, SbTPS12 was proven to be a less promiscuous class I diterpene synthase. These results give a comprehensive understanding of the terpenoid biosynthesis in S. barbata and provide useful information for enhancing the production of bioactive neo-clerodane diterpenoids through genetic engineering.

Six New neo-Clerodane Diterpenoids from Aerial Parts of Scutellaria barbata and Their Cytotoxic Activities

Planta Medica ◽  
2018 ◽  
Vol 84 (17) ◽  
pp. 1292-1299 ◽  
Author(s):  
Guo-Chun Yang ◽  
Jia-Hui Hu ◽  
Bing-Long Li ◽  
Huan Liu ◽  
Jia-Yue Wang ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Cancer ◽  
In Vitro Cytotoxicity ◽  
Cytotoxic Activities ◽  
Scutellaria Barbata ◽  
Human Cancer Cell Lines ◽  
Aerial Parts ◽  
Ic50 Values ◽  
Clerodane Diterpenoids

AbstractSix new neo-clerodane diterpenoids (1–6), scutebatas X – Z, A1-C1, along with twelve known ones (7–18) were obtained via the phytochemical investigation of the aerial parts of Scutellaria barbata. Their structures were established by detailed spectroscopic analysis. The absolute configurations of 1 and 2, as the representative members of this type, were identified based on a circular dichroic exciton chirality method. Moreover, in vitro cytotoxicity of compounds 1–6 were evaluated against three human cancer cell lines (SGC-7901, MCF-7, and A-549) using the MTT method. Compound 6 showed cytotoxic activities against all the three cell lines with IC50 values of 17.9, 29.9, and 35.7 µM, respectively.

scholarly journals Unveiling Monoterpene Biosynthesis in Taiwania cryptomerioides via Functional Characterization

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2404
Author(s):  
Li-Ting Ma ◽  
Pi-Ling Liu ◽  
Yang-Tui Cheng ◽  
Tz-Fan Shiu ◽  
Fang-Hua Chu
Keyword(s):  
Phylogenetic Analysis ◽  
Functional Characterization ◽  
In Vitro Assay ◽  
Conifer Species ◽  
Vitro Assay ◽  
Catalytic Residues ◽  
Monoterpene Synthase ◽  
Monoterpene Biosynthesis ◽  
Taiwania Cryptomerioides

Taiwania cryptomerioides is a monotypic species, and its terpenoid-rich property has been reported in recent years. To uncover monoterpene biosynthesis in T. cryptomerioides, this study used transcriptome mining to identify candidates with tentative monoterpene synthase activity. Along with the phylogenetic analysis and in vitro assay, two geraniol synthases (TcTPS13 and TcTPS14), a linalool synthase (TcTPS15), and a β-pinene synthase (TcTPS16), were functionally characterized. Via the comparison of catalytic residues, the Cys/Ser at region 1 might be crucial in determining the formation of α-pinene or β-pinene. In addition, the Cupressaceae monoterpene synthases were phylogenetically clustered together; they are unique and different from those of published conifer species. In summary, this study aimed to uncover the ambiguous monoterpenoid network in T. cryptomerioide, which would expand the landscape of monoterpene biosynthesis in Cupressaceae species.

scholarly journals Composition and Antioxidant Properties of the Essential Oil of the Endemic Cape Verdean Satureja forbesii

2009 ◽  
Vol 4 (9) ◽  
pp. 1934578X0900400 ◽  
Author(s):  
Risoleta Ortet ◽  
Erik L. Regalado ◽  
Olivier P. Thomas ◽  
Jorge A. Pino ◽  
Miguel D. Fernández ◽  
...  
Keyword(s):  
Essential Oil ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Free Radical Scavenging ◽  
In Vitro Assay ◽  
Cape Verde ◽  
Cape Verdean ◽  
Vitro Assay ◽  
Aerial Parts

The chemical composition of essential oil from the air-dried aerial parts of Satureja forbesii (Benth.) Briq. from Cape Verde was studied by GC and GC/MS. Thirty-nine volatile compounds were identified of which geranial (42.0%) and neral (31.2%) were the major constituents. Using the 2,2-diphenyl-2-picrylhydrazyl free-radical scavenging method and the in vitro assay for prevention of lipid peroxidation by thiobarbituric reactive species, significant activities were evidenced.

Halimium halimifolium: From the Chemical and Functional Characterization to a Nutraceutical Ingredient Design

Planta Medica ◽  
2019 ◽  
Vol 85 (11/12) ◽  
pp. 1024-1033 ◽  
Author(s):  
Khawla Kerbab ◽  
Francesca Sansone ◽  
Lahcene Zaiter ◽  
Tiziana Esposito ◽  
Rita Celano ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Folk Medicine ◽  
Oral Intake ◽  
In Vitro Dissolution ◽  
Bioactive Components ◽  
Ph Change ◽  
Laser Scattering ◽  
Aerial Parts ◽  
Derivatives Of

Abstract Halimium halimifolium (Hh) is a shrub used in Algerian folk medicine to treat gastrointestinal pain. An UHPLC-PDA-ESI/MSn method was developed to identify the metabolic profile of the traditionally used infusion (Hh-A) from the aerial parts. The structures of flavanols were confirmed by NMR analysis after the isolation procedure from a hydrohalcolic extract (Hh-B) that also allowed for the identification of phenolic acids, an aryl butanol glucoside, and different derivatives of quercetin, myricetin, and kaempferol. Tiliroside isomers were the chemical markers of Hh-A and Hh-B (54.33 and 36.00 mg/g, respectively). Hh-A showed a significant scavenging activity both against the radicals 1,1-diphenyl-2-picrylhydrazyl and 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (EC50 = 10.49 µg/mL and TEAC value = 1.98 mM Trolox/mg infusion) and the lipopolysaccharide-induced reactive oxygen species release in A375 and HeLa cells. Moreover, the antihyperglycemic properties, by inhibiting the α-amylase and α-glucosidase enzymes (IC50 = 0.82 mg/mL and 25.01 µg/mL, respectively), were demonstrated. To upgrade the therapeutic effect, a microencapsulation process is proposed as a strategy to optimize stability, handling, and delivery of bioactive components, avoiding the degradation and loss of the biological efficacy after oral intake. Hh-loaded microparticles were designed using cellulose acetate phthalate as the enteric coating material and spray drying as a production process. The results showed a satisfactory process yield (67.9%), encapsulation efficiency (96.7%), and micrometric characteristics of microparticles (laser-scattering, fluorescent, and scanning electron microscopy). In vitro dissolution studies (USPII-pH change method) showed that Hh-loaded microparticles are able to prevent the release and degradation of the bioactive components in the gastric tract, releasing them into the intestinal environment.

scholarly journals Chemical Constituents of Equisetum Debile and their Cytotoxic Activity

2008 ◽  
Vol 3 (11) ◽  
pp. 1934578X0800301
Author(s):  
Tran Huy Thai ◽  
Nguyen Quang Hung ◽  
Chau Van Minh ◽  
Nguyen Xuan Cuong ◽  
Pham Hai Yen ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Cancer ◽  
Chemical Constituents ◽  
In Vitro Assay ◽  
The Other ◽  
Human Cancer Cell ◽  
Vitro Assay ◽  
Human Cancer Cell Lines ◽  
Aerial Parts

A new phenyl glycoside, equisetumoside D (1), was isolated from the aerial parts of Equisetum debile, along with equisetumoside B (2), dehydrovomifoliol (3), corchoionoside C (4), (-)-isolariciresinol-3a-O-β-D-glucopyranoside (5), and kaempferol 3-O-sophoroside-7-O-β-D-glucopyranoside (6). Their structures were elucidated by NMR spectroscopic and MS experiments. Compound 1 was found to be cytotoxic against both tested human cancer cell lines, hepatocellular carcinoma (Hep-G2, IC50: 1.12 μg/mL) and rhabdosarcoma (RD, IC50: 0.25 μg/mL), while the other compounds showed no activity against these cell lines by in vitro assay.

Phytotoxic neo-clerodane diterpenoids from the aerial parts of Scutellaria barbata

2020 ◽  
Vol 171 ◽  
pp. 112230 ◽  
Author(s):  
Hang-Ying Li ◽  
Wen-Jun Wei ◽  
Kai-Liang Ma ◽  
Jie-Yao Zhang ◽  
Ya Li ◽  
...  
Keyword(s):  
Scutellaria Barbata ◽  
Aerial Parts ◽  
Clerodane Diterpenoids

scholarly journals GFP-Fragment Reassembly Screens for the Functional Characterization of Variants of Uncertain Significance in Protein Interaction Domains of the BRCA1 and BRCA2 Genes

Cancers ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 151 ◽  
Author(s):  
Laura Caleca ◽  
Mara Colombo ◽  
Thomas van Overeem Hansen ◽  
Conxi Lázaro ◽  
Siranoush Manoukian ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Functional Characterization ◽  
Brca1 And Brca2 ◽  
Vitro Assay ◽  
Variants Of Uncertain Significance ◽  
Pathogenic Variants ◽  
Brca1 And Brca2 Genes ◽  
Uncertain Significance ◽  
Genome Protection

Genetic testing for BRCA1 and BRCA2 genes has led to the identification of many unique variants of uncertain significance (VUS). Multifactorial likelihood models that predict the odds ratio for VUS in favor or against cancer causality, have been developed, but their use is conditioned by the amount of necessary data, which are difficult to obtain if a variant is rare. As an alternative, variants mapping to the coding regions can be examined using in vitro functional assays. BRCA1 and BRCA2 proteins promote genome protection by interacting with different proteins. In this study, we assessed the functional effect of two sets of variants in BRCA genes by exploiting the green fluorescent protein (GFP)-reassembly in vitro assay, which was set-up to test the BRCA1/BARD1, BRCA1/UbcH5a, and BRCA2/DSS1 interactions. Based on the findings observed for the validation panels of previously classified variants, BRCA1/UbcH5a and BRCA2/DSS1 binding assays showed 100% sensitivity and specificity in identifying pathogenic and non-pathogenic variants. While the actual efficiency of these assays in assessing the clinical significance of BRCA VUS has to be verified using larger validation panels, our results suggest that the GFP-reassembly assay is a robust method to identify variants affecting normal protein functioning and contributes to the classification of VUS.

scholarly journals The carrot monoterpene synthase gene cluster on chromosome 4 harbours genes encoding flavour-associated sabinene synthases

2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Sven Reichardt ◽  
Holger Budahn ◽  
Dominic Lamprecht ◽  
David Riewe ◽  
Detlef Ulrich ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Weight ◽  
Gene Cluster ◽  
Expression Patterns ◽  
Functional Characterization ◽  
Major Product ◽  
Terpene Synthase ◽  
Low Molecular Weight ◽  
Chromosome 4

AbstractIn plants, low molecular weight terpenes produced by terpene synthases (TPS) contribute to multiple ecologically and economically important traits. The present study investigates a carrot terpene synthase gene cluster on chromosome 4 associated with volatile monoterpene production. Two carrot mutants, yellow and cola, which are contrasting in the content of low molecular weight terpenes, were crossed to develop an F2 mapping population. The mapping analysis revealed overlapping QTLs on chromosome 4 for sabinene, α-thujene, α-terpinene, γ-terpinene, terpinen-4-ol and 4-carene. The genomic region of this locus includes a cluster of five terpene synthase genes (DcTPS04, DcTPS26, DcTPS27, DcTPS54 and DcTPS55). DcTPS04 and DcTPS54 displayed genotype- and tissue-specific variation in gene expression. Based on the QTL mapping results and the gene expression patterns, DcTPS04 and DcTPS54 were selected for functional characterization. In vitro enzyme assays showed that DcTPS54 is a single-product enzyme catalysing the formation of sabinene, whereas DcTPS04 is a multiple-product terpene synthase producing α-terpineol as a major product and four additional products including sabinene, β-limonene, β-pinene and myrcene. Furthermore, we developed a functional molecular marker that could discriminate carrot genotypes with different sabinene content in a set of 85 accessions.

scholarly journals Selective BuChE Inhibitory Activity, Chemical Composition, and Enantiomeric Content of the Essential Oil from Salvia leucantha Cav. Collected in Ecuador

Plants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1169
Author(s):  
Gabriela Villalta ◽  
Melissa Salinas ◽  
James Calva ◽  
Nicole Bec ◽  
Christian Larroque ◽  
...  
Keyword(s):  
Chemical Composition ◽  
Essential Oil ◽  
Inhibitory Activity ◽  
Enantiomeric Excess ◽  
Bornyl Acetate ◽  
Vitro Assay ◽  
Aerial Parts ◽  
Germacrene D ◽  
Enantioselective Gc

The essential oil (EO) of Salvia leucantha Cav. was isolated by steam distillation of the aerial parts collected in the South of Ecuador. Its physical properties were evaluated and the chemical composition of the oil was determined by GC-MS and GC-FID analyses using two chromatographic columns, DB-5ms and HP-INNOWax. Six major compounds were identified, namely, the sesquiterpenes 6.9-guaiadiene (19.14%), (E)-caryophyllene (16.80%), germacrene D (10.22%), (E)-β-farnesene (10.00%), and bicyclogermacrene (7.52%), and the monoterpenoid bornyl acetate (14.74%). Furthermore, four pairs of enantiomers were determined by enantioselective GC-MS of the essential oil. (−)-germacrene D and (+)-α-pinene showed the highest enantiomeric excess (ee%). In an in vitro assay, the essential oil demonstrated an interesting inhibitory activity of the enzyme butyrylcholinesterase (BuChE), with an IC50 = 32.60 µg/mL, which is the highest determined for a Salvia species. In contrast, the oil was weakly active against acetylcholinesterase (AChE) with an IC50 > 250 µg/mL.

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