A novel predict-verify strategy for targeted metabolomics: Comparison of the curcuminoids between crude and fermented turmeric.

Curcuminoids are the major bioactive constituents of turmeric, the application of which are limited by the poor bioavailability. In this study, turmeric was fermented by the Monascus purpureus and Eurotium cristatum fungi to enhance its bioavailability. To explore the variations in the curcuminoids contents in fermented turmeric, a targeted predict-verify strategy was established. For targeted analysis of curcuminoids, a compound library containing all possible curcuminoids based on their structural skeleton was predicted and built for targeted scanning. Then, the MS data were automatically matched with the predicted library to verify the corresponding curcuminoids. As a result, 115 curcuminoids (48 novel compounds and 14 compounds reported in turmeric for the first time) were fully characterized in crude and fermented turmeric. Among these curcuminoids, 31 were newly generated in fermented turmeric. The established predict-verify strategy allows for an efficient and automatic metabolomic analysis to screen for curcuminoids with potentially better bioavailability.

Drought-tolerant and plant growth-promoting endophytic Staphylococcus sp. having synergistic effect with silicate supplementation.

Endophytic bacteria have been reported to have symbiotic, mutualistic, commensalistic or trophobiotic relationships with various plant parts. As part of its adaptation, many endophytic organisms are known to exhibit properties with multiple beneficial effects to the plant system. Even though many bacterial genera have been identified to have endophytic association, isolation of those which were previously demonstrated well for human association is quite interesting. In the study, endophytic bacteria Ceb1 isolated from the rhizome of Curcuma longa was identified by 16S rDNA sequencing as Staphylococcus sp. Further, Ceb1 was observed to have the ability to tolerate drought stress. While screening for the plant growth-promoting traits, Ceb1 was found to be positive for IAA production both under drought-stressed and normal conditions as confirmed by HPLC. The Ceb1 priming with Vigna unguiculata was observed to enhance the growth parameters of the plant. Analysis of Ceb1-treated plants by ICP-MS further showed modulation of both macro- and micronutrients. Upon drought stress induction in Vigna unguiculata, Ceb1 was found to provide synergistic plant growth-promoting effect to the plant along with the supplemented silicate sources. Under the changing agroclimatic conditions, exploring the plant stress-alleviating effects of endophytes is highly significant.

Production of Gentisyl Alcohol from Phoma herbarum Endophytic in Curcuma longa L. and Its Antagonistic Activity Towards Leaf Spot Pathogen Colletotrichum gloeosporioides.

Endophytes from medicinal plants represent a potential source of bioactive compounds. During the present investigation, fungal endophytes were isolated from turmeric (Curcuma longa), an important medicinal plant. A total of 207 endophytic fungal isolates were obtained from the rhizome of C. longa L. They were grouped into seven genera based on morphological and molecular data. The fungal endophytes of C. longa were evaluated for antifungal activity against Colletotrichum gloeosporioides, the causal organism of leaf spot of turmeric. The disease is a major cause for economic loss in turmeric cultivation. Endophytic Phoma herbarum showed significant activity against C. gloeosporioides and was therefore selected for further studies. A compound gentisyl alcohol was isolated from P. herbarum which showed effective antagonism against C. gloeosporioides. The organism could therefore be used as a biocontrol agent against C. gloeosporioides.

Screening for Endophytic Fungi from Turmeric Plant (Curcuma longa L.) of Sukabumi and Cibinong with Potency as Antioxidant Compounds Producer.

Potency of medicinal plant is related to microorganisms lived in the plant tissue. Those microorganisms are known as endophytic microbes that live and form colonies in the plant tissue without harming its host. Each plant may contains several endophytic microbes that produce biological compounds or secondary metabolites due to co-evolution or genetic transfer from the host plant to endophytic microbes. Endophytic fungi research done for turmeric plant (Curcuma longa L.) gave 44 isolated fungi as results. Those 44 fungi isolated were fermented in Potato Dextrose Broth (PDB) media, filtered, extracted with ethylacetate and then were analyzed by Thin Layer Chromatography (TLC) method and tested for their antioxidant activity by radical scavenging method. The antioxidant activity of the ethylacetate filtrate extracts either from Sukabumi or Cibinong were higher than the biomass extracts. There were 6 fungi that showed antioxidant activities over 65%, i.e., with code name K.Cl.Sb.R9 (93.58%), K.Cl.Sb.A11 (81.49%), KCl.Sb.B1 (78.81%), KCl.Sb.R11 (71.67%) and K.Cl.Sb.A12 (67.76%) from Sukabumi and K.Cl.Cb.U1 (69.27%) from Cibinong. These results showed that bioproduction by endophytic microbes can gave potential antioxidant compounds.

ß-D-Glucan nanoparticle pre-treatment induce resistance against Pythium aphanidermatum infection in turmeric.

In vitro experiments were carried out to test the efficacy of GNP (ß-D-glucan nanoparticle prepared from mycelium of Pythium aphanidermatum) against rhizome rot disease of turmeric (Curcuma longa L.) caused by P. aphanidermatum. GNP (0.1%, w/v) was applied to rhizome prior to inoculation with P. aphanidermatum (0 h, 24 h). Cell death, activities of defense enzymes such as peroxidase, polyphenol oxidase, protease inhibitor and ß-1,3 glucanase were monitored. Prior application of GNP (24 h) to turmeric rhizome effectively controls P. aphanidermatum infection. The increase in defense enzyme activities occurred more rapidly and was enhanced in P. aphanidermatum infected rhizomes that were pre-treated with GNP. Pre-treatment also induced new isoforms of defense enzymes. Increased activities of defense enzymes suggest that they play a key role in restricting the development of disease symptoms in the rhizomes as evidenced by a reduction in cell death. The results demonstrated that GNP can be used as a potential agent for control of rhizome rot disease.

Essential oil content of the rhizome of Curcuma purpurascens Bl. (Temu Tis) and its antiproliferative effect on selected human carcinoma cell lines.

Curcuma purpurascens Bl., belonging to the Zingiberaceae family, is known as temu tis in Yogyakarta, Indonesia. In this study, the hydrodistilled dried ground rhizome oil was investigated for its chemical content and antiproliferative activity against selected human carcinoma cell lines (MCF7, Ca Ski, A549, HT29, and HCT116) and a normal human lung fibroblast cell line (MRC5). Results from GC-MS and GC-FID analysis of the rhizome oil of temu tis showed turmerone as the major component, followed by germacrone, ar-turmerone, germacrene-B, and curlone. The rhizome oil of temu tis exhibited strong cytotoxicity against HT29 cells (IC50 value of 4.9 ± 0.4 µg/mL), weak cytotoxicity against A549, Ca Ski, and HCT116 cells (with IC50 values of 46.3 ± 0.7, 32.5 ± 1.1, and 35.0 ± 0.3 µg/mL, resp.), and no inhibitory effect against MCF7 cells. It exhibited mild cytotoxicity against a noncancerous human lung fibroblast cell line (MRC5), with an IC50 value of 25.2 ± 2.7 µg/mL. This is the first report on the chemical composition of this rhizome's oil and its selective antiproliferative effect on HT29. The obtained data provided a basis for further investigation of the mode of cell death.

Characterization of secondary metabolites of an endophytic fungus from Curcuma wenyujin.

Endophytic fungi are ubiquitous in the plant kingdom and they produce a variety of secondary metabolites to protect plant communities and to show some potential for human use. However, secondary metabolites produced by endophytic fungi in the medicinal plant Curcuma wenyujin are sparsely explored and characterized. The aim of this study was to characterize the secondary metabolites of an active endophytic fungus. M7226, the mutant counterpart of endophytic fungus EZG0807 previously isolated from the root of C. wenyujin, was as a target strain. After fermentation, the secondary metabolites were purified using a series of purification methods including thin layer chromatography, column chromatography with silica, ODS-C18, Sephadex LH-20, and macroporous resin, and were analyzed using multiple pieces of data (UV, IR, MS, and NMR). Five compounds were isolated and identified as curcumin, cinnamic acid, 1,4-dihydroxyanthraquinone, gibberellic acid, and kaempferol. Interestingly, curcumin, one of the main active ingredients of C. wenyujin, was isolated as a secondary metabolite from a fungal endophyte for the first time.

Streptomyces phytohabitans sp. nov., a novel endophytic actinomycete isolated from medicinal plant Curcuma phaeocaulis.

A novel actinomycete, designated strain KLBMP 4601(T), was isolated from the root of the medicinal plant Curcuma phaeocaulis collected from Sichuan Province, south-west China. The strain produced extensively branched substrate and aerial hyphae that carried straight to flexuous spore chains. Chemotaxonomic properties of this strain were consistent with those of members of the genus Streptomyces. The cell wall of strain KLBMP 4601(T) contained LL-diaminopimelic acid as the characteristic diamino acid. The major menaquinone was MK-9(H(4)), with minor amounts of MK-9(H(6)), MK-9(H(8)) and MK-10(H(2)). The major fatty acids were C(16:0), iso-C(16:0), C(18:1)ω9c and C(16:1), iso G. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain KLBMP 4601(T) belongs to the genus Streptomyces and is most closely related to Streptomyces armeniacus JCM 3070(T) (97.9 %), Streptomyces pharmamarensis PM267(T) (97.6 %) and Streptomyces artemisiae YIM 63135(T) (97.5 %). The 16S rRNA gene sequence similarity between strain KLBMP 4601(T) and other members of this genus were lower than 97.5 %. DNA-DNA hybridization studies of strain KLBMP 4601(T) with the three closest species showed relatedness values of 36.3 ± 4.2 %, 27.3 ± 0.6 %, and 30.9 ± 2.5 %, respectively. On the basis of chemotaxonomic, phenotypic and genotypic characteristics, it is evident that strain KLBMP 4601(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces phytohabitans sp. nov. is proposed. The type strain is KLBMP 4601(T) (=KCTC 19892(T) = NBRC 108772(T)).

Bioactive metabolites from Chaetomium globosum L18, an endophytic fungus in the medicinal plant Curcuma wenyujin.

An endophytic fungus, strain L18, isolated from the medicinal plant Curcuma wenyujin Y.H. Chen et C. Ling was identified as Chaetomium globosum Kunze based on morphological characteristics and sequence data for the internal transcribed spacer (ITS-5.8S-ITS2) of the nuclear ribosomal DNA. A new metabolite named chaetoglobosin X (1), together with three known compounds erogosterol (2), ergosterol 5α,8-peroside (3) and 2-methyl-3-hydroxy indole (4), were isolated from C. globosum L18. Their structures were elucidated by spectroscopic methods including NMR, UV, IR and MS data and comparison with published data. Chaetoglobosin X (1) is hitherto unknown, whereas 2-methyl-3-hydroxy indole (4) is reported for the first time as a fungal metabolite, and erogosterol (2) and ergosterol 5α,8-peroside (3) are known fungal metabolites previously identified in other genera. Chaetoglobosin X (1) exhibited a broader antifungal spectrum and showed the strongest cytotoxic activity against H22 and MFC cancer cell lines.

[Studies on identification and secondary metabolites of endophytic fungi strain E8 from Curcuma wenyujin].

OBJECTIVE: To identify the endophyte strain E8 with high activity from Curcuma wenyujin and study its secondary metabolites. METHOD: The strain E8 was identified by morphological observation and ITS sequence analysis. Manifold chromatographic methods were used to separate and purify the chemical constituents of fermentation broth from strain E8, and their structures were identified by physiochemical properties and spectral data. RESULT: The strain E8 belongs to P. oxalicum. Four compounds were isolated from the fermentation broth of this strain and elucidated as chrysophanol, emodin, secalonic acid A and beta-sitosterol. CONCLUSION: The endophyte P. oxalicum was isolated from medicinal plant Curcuma wenyujin for the first time. Four compounds were first isolated from endophytic fungus in C. wenyujin. Thus, microbial fermentation is a new access for these compounds production.