A new cytochalasan compound from endophyte Chaetomium globosum WQ in Imperata cylindrical / 药学学报

italic>Chaetomium globosum WQ, an endophyte derived from Imperata cylindrical, can produce abundant cytochalasan compounds through solid state fermentation. Based on previous research and guided by 1H NMR spectrum and TLC, a new cytochalasan compound was isolated from the ethyl acetate extract of a solid culture of C. globosum WQ using silica gel column chromatography, gel filtration over Sephadex LH-20 and HPLC. The new compound was characterized as 20-iso-chaetoglobosin E (1) by a combination of spectroscopic (HR-MS, 1D and 2D NMR) analyses.

Secondary metabolites of endophyte Chaetomium globosum isolated from Hypericum sampsonii / 中草药

Objective: To study the chemical constituents of fungus Chaetomium globosum isolated from Hypericum sampsomii. Methods: By means of silica gel, Sephadex LH-20 column chromatography, and HPLC, the chemical constituents of solid culture broth of C. globosum were separated systematically. The structures of the isolated compounds were elucidated by means of physico-chemical properties and spectral data. Results: By means of spectroscopic evidences, eight compounds were isolated from the 95% ethanol extract part of the fungus C. globosum, which were identified as glolactone A (1), 4’-epialtenuene (2), altenuene (3), chaetomugilin Q (4), chaetomugilin D (5), chaetomugilide B (6), chaetoglobosin Vb (7), and chaetoglobosin C (8), respectively. Conclusion: Compound 1 is a new compound named glolactone A, and compounds 2 and 3 are isolated from this fungus for the first time.

Isolation,screening and identification of endophytic fungi and detection of its antifungal effects against Alternaria panax / 中国中药杂志

To obtain biocontrol fungus for Alternaria panax,the antifungal effects of one strain of endophytic fungi isolated from leaves of healthy ginseng were screened and evaluated by using dual-culture method,and the taxonomic assignment of the screened strain was identified based on the morphological characters and ITS sequence analysis. The results of dual-culture showed that one of the endophytes marked as FS-01 had good antifungal effects and the inhibitory rates of FS-01 strain to A. panax was( 60. 21±0. 12) %.The hyphae junction of the both strains,A. panax dissolved,broke and winded,while the hyphae of FS-01 strain remained normal. The inhibitory rates of non-sterilized FS-01 strain fermentation liqud was( 13. 94±0. 21) %. Strain FS-01 identified as Chaetomium globosum.

New production process of the antifungal chaetoglobosin A using cornstalks

Abstract Chaetoglobosin A is an antibacterial compound produced by Chaetomium globosum, with potential application as a biopesticide and cancer treatment drug. The aim of this study was to evaluate the feasibility of utilizing cornstalks to produce chaetoglobosin A by C. globosum W7 in solid-batch fermentation and to determine an optimal method for purification of the products. The output of chaetoglobosin A from the cornstalks was 0.34 mg/g, and its content in the crude extract was 4.80%. Purification conditions were optimized to increase the content of chaetoglobosin A in the crude extract, including the extract solvent, temperature, and pH value. The optimum process conditions were found to be acetone as the extractant, under room temperature, and at a pH value of 13. Under these conditions, a production process of the antifungal chaetoglobosin A was established, and the content reached 19.17%. Through further verification, cornstalks could replace crops for the production of chaetoglobosin A using this new production process. Moreover, the purified products showed great inhibition against Rhizoctonia solani, with chaetoglobosin A confirmed as the main effective constituent (IC50 = 3.88 µg/mL). Collectively, these results demonstrate the feasibility of using cornstalks to synthesize chaetoglobosin A and that the production process established in this study was effective.

Chemical constituents of liquid culture of symbiotic Chaetomium globosum ML-4 of oyster and their in vitro antitumor activity / 中国中药杂志

Isolation and purification of chemical constituents of liquid culture of symbiotic Chaetomium globosum ML-4 of oyster was performed through silica gel column chromatography, gel filtration over Sephadex LH-20, preparative TLC and HPLC. Five compounds were obtained and their structures were determined as chaetoglobosin V(1), chaetoglobosin Vb(2), tyrosol(3), 5-methyluracil(4)and uracil(5), respectively, based on HR-MS and NMR data and comparison with literatures. In vitro cytotoxicity of compounds against human hepatocellular carcinoma cell line SMMC-7721 were measured byMTT method, and results showed that compound 1 could obviously inhibit the proliferation of SMMC-7721 cells with an IC₅₀ value of 60.5 mg•L⁻¹, while the IC₅₀ value of positive control cisplatin was 19.96 mg•L⁻¹. Further studies discovered that compound 1 could lead to G2 phase arrest in SMMC-7721 cells and induce SMMC-7721 cells apoptosis. The ratio of Bcl-2/Bax in SMMC-7721 cells was decreased. The expression of protein Caspases-3,-8,-9 was improved and the expression and phosphorylation level of Akt were reduced. Aforementioned results revealed that in vitro antitumor activity of compound 1 against SMMC-7721 cells were related to G2 phase cell cycle arrest and induced-apoptosis. The induced-apoptosis was involved in both the mitochondrial pathway and the death receptor pathway and connected with activity decline of PI3K/Akt signaling pathway.

Endophytic fungus strain 28 isolated from Houttuynia cordata possesses wide-spectrum antifungal activity

Abstract The aim of this paper is to identify and investigate an endophytic fungus (strain 28) that was isolated from Houttuynia cordata Thunb, a famous and widely-used Traditional Chinese Medicine. Based on morphological methods and a phylogenetic analysis of ITS sequences, this strain was identified as Chaetomium globosum. An antifungal activity bioassay demonstrated that the crude ethyl acetate (EtOAc) extracts of strain 28 had a wide antifungal spectrum and strong antimicrobial activity, particularly against Exserohilum turcicum (Pass.) Leonard et Suggs, Botrytis cinerea persoon and Botrytis cinerea Pers. ex Fr. Furthermore, the fermentation conditions, extraction method and the heat stability of antifungal substances from strain 28 were also studied. The results showed that optimal antifungal activity can be obtained with the following parameters: using potato dextrose broth (PDB) as the base culture medium, fermentation for 4–8 d (initial pH: 7.5), followed by extraction with EtOAc. The extract was stable at temperatures up to 80 °C. This is the first report on the isolation of endophytic C. globosum from H. cordata to identify potential alternative biocontrol agents that could provide new opportunities for practical applications involving H. cordata.

HIV-1 integrase inhibitory activity of endophytic fungi from Aquilaria sinensis (Lour.) Gilg / 中国药学杂志

OBJECTIVE: To investigate the HIV-1 integrase inhibitory activity of endophytic fungi from medicinal plant Aquilaria sinensis (Lour.) Gilg. METHODS: HIV-1 integrase inhibitory activity of the isolated endophytic fungi was determined by HIV-1 integrase strand transfer inhibitory activity assay, and the active metabolites of the endophytic fungi with the greatest potential were studied by activity tracking. RESULTS: Among 78 strains of endophytic fungi isolated from A. sinensis, nine strains (11.54%) showed strong inhibitory activity against HIV-1 integrase. Three compounds were obtained from the fermentation of strain HN-AS-8 which was identified to be Chaetomium globosum by HIV-1 integrase strand reaction. Compound 1 inhibited HIV-1 integrase with IC50 value of 35.4 μmol · L-1, and compounds 1 and 3 were for the first time isolated from Chaetomium sp. CONCLUSION: Potential anti-HIV-1 metabolites exist in the endophytic fungi from A. sinensis, which can be new resources for new anti-AIDS drugs.

Cloning and expression of a small heat and salt tolerant protein (Hsp22) from Chaetomium globosum.

The present study reports molecular characterization of small heat shock protein gene in Indian isolates of Chaetomium globosum, C. perlucidum, C. reflexum, C. cochlioides and C. cupreum. Six isolates of C. globosum and other species showed a band of 630bp using specific primers. Amplified cDNA product of C. globosum (Cg 1) cloned and sequenced showed 603bp open reading frame encoding 200 amino-acids. The protein sequence had a molecular mass of 22 kDa and was therefore, named Hsp22. BlastX analysis revealed that the gene codes for a protein homologous to previously characterized Hsp22.4 gene from C. globosum (AAR36902.1, XP 001229241.1) and shared 95% identity in amino acid sequence. It also showed varying degree of similarities with small Hsp protein from Neurospora spp. (60%), Myceliophthora sp. (59%), Glomerella sp. (50%), Hypocrea sp. (52%), and Fusarium spp. (51%). This gene was further cloned into pET28a (+) and transformed E. coli BL21 cells were induced by IPTG, and the expressed protein of 30 kDa was analyzed by SDS-PAGE. The IPTG induced transformants displayed significantly greater resistance to NaCl and Na2CO3 stresses.

Effect of two kinds of endophytic fungi on salt resistance of Chrysanthemum morifolium / 中草药

Objective: The NaCl stress conditions were simulated to study the effect of the endophytic fungi C1, C4 on antisalty characteristic of Chrysanthemum morifolium in the adverse circumstance. Methods: Endophytic Botrytis sp. (C1) and Chaetomium globosum (C4) were inoculated to the C. morifolium plantlets which were planted in the pots in order to research the effects of salt stress on physiological indicators of C. morifolium. Results: With the increase of NaCl concentration, the water content of root and leaf decreased in every group. The loss of root and leaf's water in fungi-treated group was smaller than that in the control group. SOD activities in every group increased with the increase of NaCl concentration, and achieved the peak value at 20 g/L NaCl. The SOD activity in fungi-treated group was higher than that in the control group. Soluble protein of fungi-treated group was higher than that in the control group, and C4 group surpassed C1 group. POD activity increased firstly and then decreased, and compared to the control group, the POD activities in C4 and C1 groups increased by 25.50% and 1.35%, respectively at 15 g/L NaCl. PAL activity of C4 treated group was seven folds compared to the control group at 15 g/L NaCl. Conclusion: Endophytic fungi could enhance the salt-tolerant ability of C. morifolium, and the effect in C4 group was better than C1 group.

Molecular characterization of Chaetomium species using URP-PCR

Chaetomium spp. are common colonizers of soil and cellulose-containing substrates. Seventeen isolates of Chaetomium spp., which included 15 isolates of C. globosum and one each of C. reflexum and C. perlucidum, were genetically characterized with universal rice primers (URP - primers derived from DNA repeat sequences in the rice genome) using polymerase chain reaction (URP-PCR). Out of the 12 URP's used in the study, nine primers were effective in producing polymorphic fingerprint patterns from DNA of Chaetomium spp. Analysis of the entire fingerprint profile using the unweighted pair-group method with arithmetic averages (UPGMA) clearly differentiated C. globosum isolates from C. perlucidum and C. reflexum. One of the primers, URP-2R, produced a uniform DNA band of 1.9 kb in all the isolates of C. globosum but not in C. perlucidum and C. reflexum, which can be used as molecular marker to differentiate C. globosum from other species. Our results indicate that URP's are sensitive and give reproducible results for assaying the genetic variability in Chaetomium spp.

Cloning, Sequence Analysis and Proeukaryotic Expression of Peroxisomal Membrane Protein Allergen Gene from Chaetomium globosum / 中国生物工程杂志

The full length cDNA encoding peroxisomal membrane protein from Chaetomium globosum was cloned using RACE technology and the sequence in cDNA library of C. globosum in GenBank ( Accn: BP099709). The 747bp full length cDNA encoding peroxisomal membrane protein allergen (pero) gene was assembled with 412bp 3'and 508bp 5'RACE products. The open reading frame was 501 bp encoding 166 amino acids. The molecular weight of the protein was 17. 5kDa and its theoretical isoelectric point was 5.75. The pero gene was amplified using specific primers of cDNA 5'and 3'untranslated region, sequence analysis indicated that the gene have 3 exons and 2 introns. ClustalX analysis revealed that amino acids sequence of pero gene from C. globosum and Neurospora crassa shared 83% high similarity. To construct pET28a-pero expressive plasmid, pero gene was inserted into pET28a expressive vector. Escherichia coli BL21 transformed by pET28a-pero plasmid was induced with IPTG. The protein expression was analyzed with SDS-PAGE. A 21kDa pero fusion protein representing the pero gene was expressed in recombinant E. coli BL21. The sequences of cDNA,DNA and deduced amino acid of the pero gene from C. globosum were submitted to GenBank (Accn: AY555771, AY584753,AAS66898).

Descriptive Reports on Some Soil-Inhabiting Fungi in Korea

During the study of microbial structures in root-regions of tomato and red pepper from fields, various soil-inhabiting fungi were isolated with the dilution plate technique. Among them an ascomycete, Emericellopsis mirabilis and three hyphomycetes, Gliocladium solani, Humicola veronae and Verticillium chlamydosporium are presented for the first time in Korea along with Talaromyces trachyspermus, Chaetomium globosum and Doratomyces microsporus.

Phaeohyphomycosis Caused by Chaetomium Globosum: First Case Report in China / 中华皮肤科杂志

A 13 year old male farmer suffered from cutaneous and nail phaeohyphomycosis for five years is reported in this paper. The lesions were dull red, nodules and well defined plaques with ulcerations, mainly on the face, extremities, palmar and plantar surfaces and buttocks. Some of them were covered with thick greyish black crusts. General examination did not reveal abnormal findings except the skin lesions. Histopathology showed granulomatous response with numerous light brown septate branching hyphae. The colonies were greyish brown on SDA and PDA at 25℃ and 37℃ with brownish black ascomata. The ioslated strain was identified as Chaetomium globosum based on the morphological features of ascomata, ascomal hairs and ascospores.

Cloning and Characterization Analysis of 60S Ribosomal Protein L10a Gene from Chaetomium globosum / 微生物学通报

The sequence of Neurospora crassa(XP_322380)and Gibberella zeae PH-1(EAA76971)ribosomal protein gene were subjected to local tBlastn searching against the Chaetomium globosum ESTs datebase.The 765 bp full length cDNA encoding 60S ribosomal protein L10a gene was obtained.The open reading frame was 654 bp and encoded 217 amino acids.The protein molecular mass was 23.9 kD.The BlastP analysis revealed that amino acids sequence of ribosomal protein L10a gene from C.globosum shared 89% high similarity with N.crassa and 78% low similarty with Ustilago maydis.The cDNA and deduced amino acid sequence of 60S ribosomal protein L10a gene were accepted by GenBank(accession numbers: AY669070,AAT74578).