Exploration of Diverse Secondary Metabolites From Streptomyces sp. YINM00001, Using Genome Mining and One Strain Many Compounds Approach.

A talented endophytic bacteria strain YINM00001, which showed strong antimicrobial activity and multiple antibiotic resistances, was isolated from a Chinese medicinal herb Peperomia dindygulensis Miq. Phylogenetic analysis based on 16S rRNA gene sequences demonstrated that strain was closely related to Streptomyces anulatus NRRL B-2000T (99.93%). The complete genome of strain YINM00001 was sequenced. The RAxML phylogenomic tree also revealed that strain YINM00001 was steadily clustered on a branch with strain Streptomyces anulatus NRRL B-2000T under the 100 bootstrap values. The complete genome of strain YINM00001 consists of an 8,372,992 bp linear chromosome (71.72 mol% GC content) and a 317,781 bp circular plasmid (69.14 mol% GC content). Genome mining and OSMAC approach were carried out to investigate the biosynthetic potential of producing secondary metabolites. Fifty-two putative biosynthetic gene clusters of secondary metabolites were found, including the putative cycloheximide, dinactin, warkmycin, and anthracimycin biosynthetic gene clusters which consist with the strong antifungal and antibacterial activities exhibited by strain YINM00001. Two new compounds, peperodione (1) and peperophthalene (2), and 17 known compounds were isolated from different fermentation broth. Large amounts and high diversity of antimicrobial and/or anticancer compounds cycloheximide, dinactin, anthracimycin, and their analogs had been found as predicted before, which highlights strain YINM00001 as an ideal candidate for further biosynthetic studies and production improvement of these valuable compounds. Meanwhile, several gene clusters that were highly conserved in several sequenced actinomycetes but significantly different from known gene clusters might be silent under proceeding fermentation conditions. Further studies, such as heterologous expression and genetic modification, are needed to explore more novel compounds from this talented endophytic Streptomyces strain.

Novel secolignans from Peperomia dindygulensis and their inhibitory activities on JAK-STAT signaling pathways.

Nineteen secolignans (1-19), including five new ones (1-5), were isolated from the whole plant of Peperomia dindygulensis. Their structures including stereochemistry were determined by spectroscopic methods, in particular NMR and electronic CD (ECD) analysis. All the isolates were evaluated for their inhibitory activities against IFN-γ/STAT1 as well as IL-6/STAT3 signaling pathway by the method of Luciferase assay. Six 2-methene type secolignans (1, 2, 6-9) exhibited significant inhibitory activities against JAK-STAT pathways with the IC50 values both lower than 10µM.

A new tetrahydrofuran lignan from Peperomia blanda / 国际药学研究杂志

Objective To investigate the chemical constituents of Peperomia blanda. Methods Guided by HPLC detec?tion,integrated methods including vacuum liquid chromatography(VLC),ODS and semi-preparative RP-HPLC were used for separa?tion. The structure was determined by spectral analyses including ESI-MS,1D NMR,2D NMR and ECD spectra. Results Four com?pounds were isolated and identified as(7S,7′S,8R,8′R)-7-(5-methoxy-3,4-methylenedioxyphenyl)-7′-(4-hydroxy-3,5-dimethoxy?phenyl)-8,8′-dihydroxymethyltetrahydrofuran(1),7,8-trans-8,8′-trans-7′,8′-cis-7,7′-(4-hydroxy-3,5-dimethoxy phenyl)-8,8′-di?acetoxymethyltetrahydrofuran(2),(+)-(7S,7′S,8R,8′R)-4,4′-dihydroxy-3,3′,5,5′-tetramethoxy-7,9′,7′,9-diepoxylignane(3), and(6R,7E,9R)-9-hydroxy-4,7-megastigmadien-3-one(4). Conclusion Compound 1 is a new tetrahydrofuran lignan,compounds 2-4 were isolated from P. blanda for the first time.

A new tetrahydrofuran lignan from Peperomia blanda / 国际药学研究杂志

Objective To investigate the chemical constituents of Peperomia blanda. Methods Guided by HPLC detection, integrated methods including vacuum liquid chromatography (VLC), ODS and semi-preparative RP-HPLC were used for separation. The structure was determined by spectral analyses including ESI-MS, 1D NMR, 2D NMR and ECD spectra. Results Four compounds were isolated and identified as (7S,7’S,8R,8 ’R)-7-(5-methoxy-3,4-methylenedioxyphenyl)-7-(4-hydroxy-3,5-dimethoxy-phenyl)-8, 8-dihydroxymethyltetrahydrofuran (1), 7, 8-trans-8,8-trans-7,8-cis-7, 7-(4-hydroxy-3,5-dimethoxy phenyl)-8,8-di-acetoxymethyltetrahydrofuran (2), (+)-(7S, 7S, 8R, 8R)-4, 4-dihydroxy-3, 3, 5, 5-tetramethoxy-7, 9, 7, 9-diepoxylignane (3), and (6R, 7E, 9R)-9-hydroxy-4, 7-megastigmadien-3-one (4). Conclusion Compound 1 is a new tetrahydrofuran lignan, compounds 2-4 were isolated from P. blanda for the first time.

Qualitative and quantitative analyses of bioactive secolignans from folk medicinal plant Peperomia dindygulensis using UHPLC-UV/Q-TOF-MS.

Peperomia dindygulensis, with secolignans (SLs) as major bioactive constituents, is a commonly used traditional folk medicine in mainland China for treatment of stomach, liver, mammary, and esophageal cancers. However, to date, there is no method available for the qualitative and quantitative analyses of SLs in this medicinal plant. The purpose of this study was to establish a sensitive, selective, and reproducible method for rapidly profiling, identifying, and determining SLs in the whole plant of P. dindygulensis. Ultra high-performance liquid chromatography (UHPLC) coupled with ultraviolet detector (UV) and quadrupole tandem time-of-flight mass spectrometry (Q-TOF-MS) were used for this analyses. The fragmentation behaviors of different types of SLs were described. A total of thirteen SLs, including two new derivatives, were identified or tentatively characterized in P. dindygulensis samples. In addition, seven major SLs in herbal samples from different regions in China were successfully determined. The method developed in this study is suitable for the qualitative and quantitative analyses of SLs in P. dindygulensis, and may be applicable for determining or identifying SLs from other Pepermia genus plants.

Simultaneous determination of five bioactive secolignans in rat plasma by LC-MS/MS for pharmacokinetic studies following oral administration of Peperomia dindygulensis Miq. extract.

A rapid and sensitive ultra fast performance liquid chromatography-tandem mass spectrometry method was developed for the simultaneous determination of five bioactive secolignans in Peperomia dindygulensis extract, including peperomin A, peperomin B, peperomin C, 4″-hydroxypeperomin B and 4″-hydroxypeperomin C in rat plasma. Arctigenin was used as the internal standard. The separation was performed on an Innovation™ Polar-RP C18 column by a gradient elution within a runtime of 7min. The mobile phase consisted of A (methanol) and B (0.1% formic acid in water) at a flow rate of 0.4mL/min. The detection was accomplished by using positive ion TurboIonSpray ionization in multiple reaction monitoring mode. The method was linear for all analytes over investigated range with all correlation coefficients greater than 0.9972. The lower limits of quantification were 1.1ng/mL for peperomin A, 1.24ng/mL for peperomin B, 1.02ng/mL for peperomin C, 1.91ng/mL for 4″-hydroxypeperomin B and 1.27ng/mL for 4″-hydroxypeperomin C. The intra- and inter-day precision (RSD%) was within 15% and the accuracy (RE%) ranged from -11.7% to 10.3%. This simple and sensitive method was fully validated and successfully applied to the pharmacokinetic study of peperomin A, peperomin B, peperomin C, 4″-hydroxypeperomin B and 4″-hydroxypeperomin C in rat plasma after oral administration of P. dindygulensis extract.

Two novel polyketides from Peperomia dindygulensis / 中草药

Objective: To study the chemical constituents from Peperomia dindygulensis. Methods: The chemical constituents in chloroform fraction of ethanol extract from P. dindygulensis were isolated and purified by column chromatography over silica gel column, preparative TLC, and semi-preparative HPLC. Their chemical structures were elucidated on the basis of physicochemical and spectral data. Results: Two new polyketides were isolated from the chloroform extracting fraction of P. dindygulensis and identified as (4S)-1,4-dihydroxy-2-(1′, 13′-diketone-octadec-14′E-ene)-1-cyclohexen-3-one (1) and (4S)-1,4-dihydroxy-2-(1′,14′-diketone-octadec-12′E-ene) -1-cyclohexen-3-one (2). Conclusion: Compounds 1 and 2 are new compounds and named as peperomadinone A and peperomadinone B.

Chemical constituents of Peperomia dindygulensis / 中草药

Objective To study the chemical constituents of Peperomia dindygulensis. Methods Chromatography was used to isolate and purify the chemical constituents, their structures were identified by spectral analyses. Results Eight compounds were isolated and identified as bis-(2-methoxy-4, 5-methylenedioxy)-benzophenone (Ⅰ), peperomin B (Ⅱ), peperomin C (Ⅲ), 5-hydroxy-4′, 7, 8-trimethoxy flavone (Ⅳ), 5-hydroxy-3′, 4′, 7, 8-tetramethoxy flavone (Ⅴ), 5, 3′-dihydroxy-4′, 7, 8-trimethoxy flavone (Ⅵ), ?-sitosterol (Ⅶ), hexadecanoic acid (Ⅷ). Conclusion Compound Ⅰ is a new compound named as dindygulensin. All compounds, except Ⅴ, are isolated from P. dindygulensis for the first time.