Comprehensive untargeted lipidomics study of black morel (Morchella sextelata) at different growth stages.

The black morel (Morchella sextelata) is a valuable edible and medicinal mushroom appreciated worldwide. Here, lipidomic profiles and lipid dynamic changes during the growth of M. sexletata were analyzed using ultra-performance liquid chromatography coupled with mass spectrometry. 203 lipid molecules, including four categories and fourteen subclasses, were identified in mature fruiting bodies, with triacylglycerol being the most abundant (37.00 %). Fatty acid composition analysis revealed that linoleic acid was the major fatty acid among the free fatty acids, glycerolipids and glycerophospholipids. The relative concentration of lipids in M. sextelata changed significantly during its growth, from which 12 and 29 differential lipid molecules were screened out, respectively. Pathway analysis based on these differential lipids showed that glycerophospholipid metabolism was the major pathway involved in the growth of M. sextelata. Our study provides a comprehensive understanding of the lipids in M. sextelata and will facilitate the development and utilization of M. sextelata.

Effect of vanadium and tungsten loading order on the denitration performance of F-doped V2O5-WO3/TiO2 catalysts.

F-doped V2O5-WO3/TiO2 catalyst has been confirmed to have excellent denitration activity at low temperatures. Since the V2O5-WO3/TiO2 catalyst is a structure-sensitive catalyst, the loading order of V2O5 and WO3 may affect its denitration performance. In this paper, a series of F-doped V2O5-WO3/TiO2 catalysts with different V2O5 and WO3 loading orders were synthesized to investigate the effect of denitration performance at low temperatures. It was found that the loading orders led to significant gaps in denitration performance in the range of 120-240 °C. The results indicated loading WO3 first better utilized the oxygen vacancies on the TiF carrier promoting the generation of reduced vanadium species. In addition, loading WO3 first facilitated the dispersion of V2O5 thus enhanced the NH3 adsorption capacity of VWTiF. In situ DRIFT verified the rapid reaction between NO2, nitrate, and nitrite species and adsorbed NH3 over the VWTiF, confirming that the NH3 selective catalytic reduction (NH3-SCR) reaction over VWTiF at 240 °C proceeded by the Langmuir-Hinshelwood (L-H) mechanism. This research established the constitutive relationship between the loading order of V2O5 and WO3 and the denitration performance of the F-doped VWTi catalyst providing insights into the catalyst design process.

Secondary Metabolites from Hericium erinaceus and Their Anti-Inflammatory Activities.

Hericium erinaceus, a culinary and medicinal mushroom, is widely consumed in Asian countries. Chemical investigation on the fruiting bodies of Hericium erinaceus led to the isolation of one new ergostane-type sterol fatty acid ester, erinarol K (1); and eleven known compounds: 5α,8α -epidioxyergosta-6,22-dien-3ß-yl linoleate (2); ethyl linoleate (3); linoleic acid (4); hericene A (5); hericene D (6); hericene E (7); ergosta-4,6,8(14),22-tetraen-3-one (8); hericenone F (9); ergosterol (10); ergosterol peroxide (11); 3ß,5α,6α,22E-ergosta-7,22-diene-3,5,6-triol 6-oleate (12). The chemical structures of the compounds were determined by 1D and 2D NMR (nuclear magnetic resonance) spectroscopy, mass spectra, etc. Anti-inflammatory effects of the isolated aromatic compounds (5-7, 9) were evaluated in terms of inhibition of pro-inflammatory mediator (TNF-α, IL-6 and NO) production in lipopolysaccharide (LPS)-stimulated murine RAW 264.7 macrophage cells. The results showed that compounds 5 and 9 exhibited moderate activity against TNF-α (IC50: 78.50 µM and 62.46 µM), IL-6 (IC50: 56.33 µM and 48.50 µM) and NO (IC50: 87.31 µM and 76.16 µM) secretion. These results supply new information about the secondary metabolites of Hericium erinaceus and their anti-inflammatory effects.

Untargeted Metabolomics Reveals Alterations in the Primary Metabolites and Potential Pathways in the Vegetative Growth of Morchella sextelata.

Morchella sextelata, one of the true morels, has recently been artificially cultivated with stable production in China. Analysis of the variations in primary metabolites during the vegetative stages of M. sextelata is critical for understanding the metabolic process. In this study, three developmental stages were categorized based on morphological and developmental changes, including the young mushroom period, marketable mature period, and physiological maturity period. Untargeted metabolomics-based mass spectrometry was used to analyze the change of metabolites during the growth stages of M. sextelata. The result showed that the metabolites' content at the different growth stages were significantly different. The relative contents of linoleic acid, mannitol, oleamide, and betaine were higher at each growth stage. Flavor substances were significantly metabolizable during commodity maturity, while amino acids, organic acids, and lipids were significantly metabolizing at physiological maturity. Pathway analysis of the most significant changes involved Pyrimidine metabolism, Vitamin B6 metabolism, Arginine biosynthesis, Lysine biosynthesis, and Lysine degradation. The results can provide a theoretical basis for further clarifying the metabolic regulation mechanism and lay the foundation for optimizing the cultivation process of M. sextelata.

Integration of Transcriptomics and Metabolomics for Understanding the Different Vegetative Growth in Morchella Sextelata.

Morchella sextelata is an edible and medicinal fungus with high nutritional, medicinal, and economic value. Recently, M. sextelata has been produced through artificial cultivation in China, but its stable production remains problematic because the details of its growth and development process are limitedly understood. Herein, to investigate the dynamic process of M. sextelata development, we integrated the transcriptomics and metabolomics data of M. sextelata from three developmental stages: the young mushroom period (YMP), marketable mature period (MMP), and physiological maturity period (PMP). The results showed that the transcriptome changed dynamically at different stages and demonstrated the significant enrichment of pathways that regulate plant growth and development, such as N-glycan biosynthesis and carbon and purine metabolism. Similarly, small-molecule metabolites, such as D-fructose-1,6-biphosphate, which was upregulated during the YMP, dihydromyricetin, which was upregulated during the MMP, and L-citrulline, which was upregulated during the PMP, also showed phase-dependent characteristics. Then, combined analysis of the transcriptome data and metabolome traits revealed that the transcriptome may affect metabolic molecules during different growth stages of M. sextelata via specific enzymes, such as α-glucosidase and glucanase, which were included in two opposite transcriptome modules. In summary, this integration of transcriptomics and metabolomics data for understanding the vegetative growth of M. sextelata during different developmental stages implicated several key genes, metabolites, and pathways involved in the vegetative growth. We believe that these findings will provide comprehensive insights into the dynamic process of growth and development in M. sextelata and new clues for optimizing the methods for its cultivation application.

Chemical Composition of Aromas and Lipophilic Extracts from Black Morel (Morchella importuna) Grown in China.

Morels (Morchella spp.) are valuable medicinal and edible mushrooms. In this study, chemical profiles of aromas and lipophilic extracts of black morel (Morchella importuna) grown in China were analyzed by gas chromatography/mass spectroscopy, along with the evaluation of antioxidant and antimicrobial activities for the lipophilic extracts. Sixty-five compounds in total were identified from the aromas, and 1-octen-3-ol was the main component for aromas of fresh (34.40%) and freeze-dried (68.61%) black morels, while the most abundant compound for the aroma of the oven-dried sample was 2(5H)-furanone (13.95%). From the lipophilic extracts, 29 compounds were identified with linoleic acid as the main compound for fresh (77.37%) and freeze-dried (56.46%) black morels and steroids (92.41%) as the main constituent for an oven-dried sample. All three lipophilic extracts showed moderate antioxidant activities against 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) with the IC50 values ranging 7.56 ∼ 17.52 mg/mL and 5.75 ∼ 9.73 mg/mL, respectively, and no obvious antimicrobial activity was observed for lipophilic extracts. The drying methods affect the chemical profile of black morel, and freeze-drying was favorable for retaining nutrients and morel smell. This is the first report on the aroma and lipophilic extracts of M. importuna grown in China.

Chemical Composition and Antimicrobial Activity of Essential Oil from Phytolacca dodecandra Collected in Ethiopia.

The essential oil from Phytolacca dodecandra, a traditional herb of Ethiopia, has been studied, including the chemical composition and antimicrobial activity. The difference between four P. dodecandra samples (P-1⁻P-4), which differed in gender or location, has also been analyzed. The essential oils were obtained by steam distillation, while the aromas were extracted by head space solid-phase microextraction (HS-SPME) and both were analyzed by gas chromatography- mass spectrometry (GC-MS). The oils' antimicrobial activities were evaluated by the microdilution method against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Candida albicans. Ninety one components, representing 88.37 to 94.01% of the aromas, were identified. The compositions of the aromas of four samples are mainly dominated by aldehydes and ketones: 2-nonanone (1.80⁻30.80%), benzaldehyde (4.99⁻25.99%), and sulcatone (2.34⁻5.87%). Sixty components representing 64.61 to 69.64% of the oils were identified, and phytone (3.04⁻21.23%), phytol (4.11⁻26.29%) and palmitic acid (1.49⁻23.87%) are the major compounds. No obvious antimicrobial activity was observed for all the four essential oils.

Biochemical and Structural Characterization of TtnD, a Prenylated FMN-Dependent Decarboxylase from the Tautomycetin Biosynthetic Pathway.

Tautomycetin (TTN) is a polyketide natural product featuring a terminal alkene. Functional characterization of the genes within the ttn gene cluster from Streptomyces griseochromogenes established the biosynthesis of the TTN polyketide backbone, its dialkylmaleic anhydride moiety, the coupling of the two moieties to form the nascent intermediate TTN F-1, and the tailoring steps converting TTN F-1 to TTN. Here, we report biochemical and structural characterization of TtnD, a prenylated FMN (prFMN)-dependent decarboxylase belonging to the UbiD family that catalyzes the penultimate step of TTN biosynthesis. TtnD catalyzes decarboxylation of TTN D-1 to TTN I-1, utilizing prFMN as a cofactor generated by the TtnC flavin prenyltransferase; both TtnD and TtnC are encoded within the ttn biosynthetic gene cluster. TtnD exhibits substrate promiscuity but accepts only TTN D-1 congeners that feature an α,ß-unsaturated acid, supporting the [3+2] cycloaddition mechanism during catalysis that requires the double bond of an α,ß-unsaturated acid substrate. TtnD shares a similar overall structure with other members of the UbiD family but forms a homotetramer in solution. Each protomer is composed of three domains with the active site located between the middle and C-terminal domains; R169-E272-E277, constituting the catalytic triad, and E228, involved in Mn(II)-mediated binding of prFMN, were confirmed by site-directed mutagenesis. TtnD represents the first example of a prFMN-dependent decarboxylase involved in polyketide biosynthesis, expanding the substrate scope of the UbiD family of decarboxylases beyond simple aromatic and cinnamic acids. TtnD and its homologues are widespread in nature and could be exploited as biocatalysts for organic synthesis.

Cytotoxic sesquiterpenoids from Ligularia pleurocaulis.

The phytochemical investigation of whole plants of Ligularia pleurocaulis led to isolation of five sesquiterpenoids, including an eremophilenolide dimer named biligupleurolide, along with eight known sesquiterpenoids. Their structures were determined by spectroscopic methods including 2D NMR techniques, and the structures of biligupleurolide and ligupleurolide were confirmed by X-ray diffraction analysis. All of the isolated compounds were tested for their cytotoxic activity against three cancer cell lines (HepG2, A549 and MCF-7), and biligupleurolide showed moderate inhibitory activities, with IC50 values of 5.2, 5.6 and 8.4 µM against MCF-7, HepG2 and A549, respectively.

Terpenoids from Ligularia kangtingensis.

BACKGROUND: Ligularia kangtingensis, a species from the genus Ligularia (Compositae), is an indigenous plant in Southwest China and more than 20 species in this genus have been used as folk medicines in China. OBJECTIVE: The chemical constituents of the whole plant of L. kangtingensis were studied. MATERIALS AND METHODS: The dried whole plants were extracted with ethanol. Its chemical constituents were mainly isolated and purified by silica gel and Sephadex LH-20 column chromatography and their structures were identified on the basis of spectral analysis. RESULTS: Twelve known terpenoids, including two monoterpenoids, five sesquiterpenoids and five triterpenoids, were isolated and identified from the whole plant of L. kangtingensis. CONCLUSION: All of the 12 known compounds were isolated for the first time from L. kangtingensis.

Strain prioritization for natural product discovery by a high-throughput real-time PCR method.

Natural products offer unmatched chemical and structural diversity compared to other small-molecule libraries, but traditional natural product discovery programs are not sustainable, demanding too much time, effort, and resources. Here we report a strain prioritization method for natural product discovery. Central to the method is the application of real-time PCR, targeting genes characteristic to the biosynthetic machinery of natural products with distinct scaffolds in a high-throughput format. The practicality and effectiveness of the method were showcased by prioritizing 1911 actinomycete strains for diterpenoid discovery. A total of 488 potential diterpenoid producers were identified, among which six were confirmed as platensimycin and platencin dual producers and one as a viguiepinol and oxaloterpin producer. While the method as described is most appropriate to prioritize strains for discovering specific natural products, variations of this method should be applicable to the discovery of other classes of natural products. Applications of genome sequencing and genome mining to the high-priority strains could essentially eliminate the chance elements from traditional discovery programs and fundamentally change how natural products are discovered.

Structural and stereochemical studies of a tetralin norsesquiterpenoid from Ligularia kangtingensis.

A chiral tetralin norsesquiterpenoid, ligukangtinol, was isolated from the plant Ligularia kangtingensis. Its planar structure was determined by extensive analysis of spectroscopic data (MS, IR, and NMR), and (1S,3R) absolute configuration of the tetralin ring was established by TDDFT-ECD calculations of the solution conformers. Conformational analysis and ECD calculations proved that the semiempirical helicity rules of 6-hydroxytetralins correlating the (1)L(b) Cotton effect and P/M helicity of the fused carbocyclic ring correctly predicts the absolute configuration and thus can be used for the configurational assignment of related substituted tetralin derivatives.

Sibiralactone: a new monoterpene from Sibiraea angustata.

Sibiralactone (1), a new monoterpene derivative, was isolated from the leaves of Sibiraea angustata. The structure was determined by the analysis of its NMR data and the absolute configuration was established by TDDFT ECD calculation of the solution conformers.

Triterpenoid glycosides from the leaves of Ilex pernyi.

Ten triterpenoid glycosides, including five new ones (1-5), were isolated from the leaves of Ilex pernyi. The chemical structures of 1-5 were determined on the basis of the chemical and spectroscopic evidence.

An effective O-demethylation of some C19-diterpenoid alkaloids with HBr-glacial acetic acid.

The aconitine-type alkaloids talatisamine (1), 8,14-diacetyltalatisamine (11), and compound 3, the lycoctonine-type alkaloid deltaline (5), and the 7,17-seco C(19)-diterpenoid alkaloids 7 and 9 were treated with HBr-glacial acetic acid to give useful O-demethylated derivatives 2, 2, 4, 6, 8, and 10 respectively in good to high yields (49-90%).

Chemical constituents from the leaves of Ilex pernyi.

A new compound and five known compounds were isolated from the ethanolic extract of the leaves of Ilex pernyi Franch. Their structures were established on the basis of spectral analysis and identified as trans-isoeugenyl-alpha-L-arabinopynosyl (1 --> 6) -beta-D-glucopyranoside (1) , kaempferol-3-O-sambubioside (2), quercetin-3-O-sambubioside (3), isoquercitrin (4), (+) -syringaresinol-O-beta-D-glucopyranoside (5), amarantholidoside IV (6). Among them, compound 1 is a new phenolic glycoside, named as ilexperphenoside A, and compounds 2-6 were isolated from this plant for the first time.

[Studies on triterpenoid constituents in leaf of Ilex pernyi].

OBJECTIVE: To investigate the chemical constituents of Ilex pernyi. METHOD: The chemical constituents were isolated by various column chromatographic methods. The structures were identified by spectral data. RESULT: Eight triterpenoid compounds were isolated and identified as ursolic acid (1), lupeol (2), alpha-amyrin (3), uvaol (4), 3beta-hydroxyurs-11-ene-13beta-olide (5), pomolic acid (6), lup-20 (29)-ene-3beta, 24-diol (7), 3beta, 23-dihydroxy-urs-12-en-28-oic acid (8). CONCLUSION: The eight compounds were obtained from this plant for the first time.

Complete assignments of (1)H and (13)C NMR spectroscopic data for two new triterpenoid saponins from Ilex pernyi.

Ilexpernoside A and ilexpernoside B, two new pentacyclic C(4)-nortriterpenoid saponins, were isolated from the leaves of Ilex pernyi Franch. Their chemical structures were determined by MS, NMR spectroscopy and chemical analysis. Complete assignments of the (1)H and (13)C NMR spectroscopic data were achieved by 1D and 2D NMR experiments (HSQC, HMBC, (1)H-(1)H COSY and NOESY).