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Natural products have high structural diversity with various pharmacological or biological activities, which are of great significance to our life and drug research. Millions of natural products with versatile structural diversity have been found in nature. In recent years, a large number of microbial genome sequences have been released in public databases and revealed many silent or cryptic secondary metabolite gene clusters hidden in their genomes. This shows the great potential for discovering new metabolites. Advances in sequencing technology and bioinformatics analysis also provide great advantages for studying the biosynthesis and structural diversity of these metabolites. Structural differentiation of natural products begins with the formation of basic scaffolds using basic building blocks derived from primary metabolism catalyzed by different backbone enzymes. The structural complexity of natural products mainly arises from tailoring enzymes to highly functionalize the skeletons with a set of chemical transformations. The well-studied modification enzymes range from different types of oxidoreductases, cytochrome P450 enzymes, to various prenyltransferases (PTs) and methyltransferases (MTs). In addition, nonenzymatic events have also contributed to the formation of final products with vast diversity and complexity. Therefore, fully exploring these unexplored gene clusters and the substrate promiscuity of enzymatic and non-enzymatic reactions for the natural product formation may be a promising way and a new strategy to explore the metabolite diversity. In a cooperation study with Dr. Liujuan Zheng, the biosynthesis of a highly oxygenated phenethyl derivative ustethylin A, isolated from Aspergillus ustus, was elucidated. Due to the instability of ustethylin A, it was acetylated before isolation and structure elucidation. Gene deletion and heterologous expression proved that the phenethyl core structure is assembled by a polyketide synthase (UttA) harboring a methyltransferase domain. Isotopic labelling experiments proved that the backbone of ustethylin A is derived from malonyl-CoA and the methyl groups, also in the phenethyl residue, are from L-methionine. Modifications on the core structure by an aryl acid reductase (UttJ), a putative nonheme FeII/2-oxoglutarate dependent oxygenase (UttH), a cytochrome P450 enzyme (UttC) and a O-methyltransferase (UttF) led to the final product ustethylin A. This study is the first report on the biosynthetic pathway of a phenethyl-containing natural product. In cooperation with Dr. Jing Liu, the biosynthesis of streptoazine C and guanitrypmycin D1 was elucidated. Firstly, a three-gene cluster coding for a cyclodipeptide synthase, a prenyltransferase, and a methyltransferase was identified in Streptomyces aurantiacus by genome mining. Heterologous expression and precursor incubation experiments led to the elucidation of the biosynthetic steps of streptoazine C. In vivo biotransformation experiments proved the high flexibility of the prenyltransferase SasB toward tryptophan-containing cyclodipeptides and their dehydroderivatives for regular C-3-prenylation. This study provides an enzyme with a high substrate promiscuity from the less explored prenyltransferase group in cyclodipeptide synthase-related pathways. Afterwards, a two-gene cluster coding for a CDPS and a cytochrome P450 was identified in Streptomyces sp. NRRL S-1521 by phylogenetic analysis. Heterologous expression and structural elucidation of the isolated products proved that the cytochrome P450 GutD1521 catalyzes the regiospecific transfer of guanine to C-2 of the indole ring of cyclo-(L-Trp-L-Tyr) via a C−C linkage, which represents a new chemical transformation within this enzyme class. Precursor incubation experiments revealed that GutD1521 efficiently accepts several other tryptophan-containing cyclodipeptides or derivatives for regiospecific coupling with guanine, thus generating different guanitrypmycin analogs. This study provides a biocatalyst for a new linkage pattern between a indole ring and a guanine moiety and expands the functional spectrum of P450s as tailoring enzymes.

The use of microorganisms for remediation and restoration of hydrocarbons contaminated soils is an effective and economic solution. The current study aims to find out efficient telluric filamentous fungi to degrade petroleum hydrocarbons pollutants. Six fungal strains were isolated from used engine (UE) oil contaminated soil. Fungi were screened for their ability to degrade crude oil, diesel and UE oil using 2.6-dichlorophenol indophenol (DCPIP). Two isolates were selected, identified and registered at NCBI as Aspergillus ustus HM3.aaa and Purpureocillium lilacinum HM4.aaa. Fungi were tested for their tolerance to different concentration of petroleum oils using radial growth diameter assay. Hydrocarbons removal percentage was evaluated gravimetrically. The degradation kinetic of crude oil was studied at a time interval of 10 days. A.ustus was the most tolerant fungi to high concentration of petroleum oils in solid medium. Quantitative analysis showed that crude oil was the most degraded oil by both isolate; P. lilacinium and A. ustus removed 44.55% and 30.43% of crude oil, respectively. The two fungi were able to degrade, respectively, 27.66 and 21.27% of diesel and 14.39 and 16.00% of UE oil. As compared to the controls, these fungi accumulated high biomass in liquid medium with all petroleum oils. Likewise, crude oil removal rate constant (K) and half-lives (t1/2) were 0.02 day−1, 34.66 day and 0.015 day−1, 46.21 day for P. lilacinium and A. ustus, respectively. The selected fungi appear interesting for petroleum oils biodegradation and their application for soil bioremediation require scale-up studies.

A new, nonglycosylated, serine proteinase capable of hydrolyzing collagen, elastin, and fibrin was isolated from a complex preparation of proteins formed by the micromycete A. ustus 1 under solid-state fermentation. The enzyme had a molecular weight of 33 kDa and a pI of 4.6. The pH-optimum (6.0) and temperature optimum (41°C) of the work of this proteinase were also determined.

Accumulation of two benzoyl esters in Aspergillus ustus after feeding with alcohols was reported 30 years ago. To the best of our knowledge, the biosynthesis for these esters has not been elucidated prior to this study. Here, we demonstrate that these compounds are artifical products of the phenethyl polyketide ustethylin A biosynthestic pathway. In addition, four aditional benzoyl esters with different methylation levels were also isolated and identified as shunt products. Feeding experiments provided evidence that the enzyme-bound polyketide acyl intermediates are hijacked by externally fed MeOH or EtOH, leading to the formation of the benzoyl esters. Graphic abstract

Genome mining of Aspergillus ustus 094102 enabled the discovery of a multiproduct bifunctional terpene synthase (BTS), AuAS. Heterologous expression of AuAS led to the discovery of five new sesterterpenes, and coexpression of the upstream CYP450 monooxygenase (AuAP450) generated four new sesterterpene alcohols. Additionally, aspergilol A showed cytotoxic activities against MCF-7, MDA-MB231, and HepG2 cancer cells (IC50 21.20-48.76 μM), and aspergilol B exhibited a cytotoxic effect on MCF-7 cells (IC50 27.41 μM).

Bioactivity-guided isolation of Aspergillus ustus led to the discovery of five new drimane sesquiterpenes, named ustusal A, ustusolate F and G, and ustusoic acid A and B, 1-5 respectively. Structural elucidation of these fungal terpenes relied on 1D and 2D NMR techniques, high-resolution mass spectrometry, and chiroptical properties. Their relative configurations were determined by NMR methods, while the absolute configurations were established using comparative analyses of computed and experimental NMR chemical shifts and ECD spectra. The sesquiterpenes exhibited weak activity against the clinically relevant pathogens vancomycin-resistant Enterococcus faecium and multidrug-resistant Staphylococcus aureus; however, the activity of 5 was drastically enhanced when equal amounts of stromemycin (6), a known metabolite coisolated from the same fraction from A. ustus, was added.

A highly oxygenated phenethyl derivative ustethylin A was isolated from Aspergillus ustus. Gene deletion, isotope labeling, and heterologous expression proved that the phenethyl core structure is assembled from malonyl-CoA by a polyketide synthase harboring a methyltransferase domain. Propionate was converted via acetyl-CoA to malonyl-CoA and incorporated into the molecule. Modifications on the core structure by three different oxidoreductases and one O-methyltransferase lead to the final product, ustethylin A.

One new dihydrobenzofuran derivative (1), known depsipeptide emericellamide A (2), three known drimanes (3–5) and two artifact drimane derivatives (6, 7) were isolated from the marine-derived fungu...

In the current study, new biomasses were prepared from fungi isolates including, Penicillium chrysogenum and Aspergillus ustus and utilized for the removal of heavy metals. During the study, the in...

New antifungal agrochemicals, derived from 2-(benzylsulfonyl)benzothiazole were synthesized by an environmentally friendly method, using water as reaction medium. These compounds were prepared by a one-pot, two-step synthesis, starting from 2-mercaptobenzothiazole and benzyl halides. The potential fungicides were tested against a panel of phytopathogenic fungi, with many of them showing a significant improvement compared to the non-oxidized analogues and the commercial antifungal Captan. The new derivatives 2-((2-chlorobenzyl)sulfonyl)benzo[d]thiazole (4f) and 2-((4-methylbenzyl)sulfonyl)benzo[d]thiazole (4k) presented remarkable properties, being able to inhibit the growth of two resistant moulds (Aspergillus fumigatus and Aspergillus ustus). Both 4f and 4k could be classified as broad-spectrum fungicides, emerging as possible candidates for the control of these moulds, which have negative impact in food production. Fil: Ballari, María Sol. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina Fil: Herrera Cano, Natividad Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina. Universidad Nacional de San Juan. Facultad de Ingeniería. Instituto de Biotecnología; Argentina Fil: Wunderlin, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Ciencia y Tecnología de Alimentos Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Ciencia y Tecnología de Alimentos Córdoba; Argentina Fil: Feresin, Gabriela Egly. Universidad Nacional de San Juan. Facultad de Ingeniería. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Santiago, Ana Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina

Oxepinamides are fungal oxepine-pyrimidinone-ketopiperazine derivatives. In this study, we elucidated the biosynthetic pathway of oxepinamide D in Aspergillus ustus by gene deletion, heterologous expression, feeding experiments, and enzyme assays. We demonstrated that the cytochrome P450 enzymes catalyzed highly specific and stereoselective oxepin ring formation.

Aspergillus sections Usti and Cavernicolarum are accommodated in the subgenus Nidulantes. In the present study, a polyphasic approach using morphology and multi-gene phylogeny was applied to investigate the taxonomy of these two sections. Based on the phylogenetic analysis, Aspergillus section Usti includes 25 species, which can be assigned to four series: Calidousti, Deflecti, Monodiorum and Usti. Aspergillus sigarelli is newly described in this section and this species was isolated from a cigarette from PR China and belongs to series Calidousti. It is clearly distinct from other members in this series based on ITS, BenA, CaM and RPB2 sequences. Aspergillus section Usti members like A. calidoustus and A. granulosus are important opportunistic pathogens, it is speculative that more pathogenetic species will be found by using polyphasic taxonomy approaches. Aspergillus section Cavernicolarum includes five species, the growth rates on agar media and size and ornamentation of conidia are important characters for differentiating species in section Cavernicolarum.

The Aspergilli of section Usti (group ustus) are represented by over 20 species, of which Aspergillus calidoustus is the most relevant human pathogen. Invasive aspergillosis (IA) caused by these fungi is rare but could represent an emerging issue among the expanding population of patients with long-term immunosuppression receiving antifungal prophylaxis. Clinicians should be aware of this unusual type of IA, which often exhibits distinct clinical features, such as an insidious and prolonged course and a high occurrence of extra-pulmonary manifestations, such as skin/soft tissue or brain lesions. Moreover, these Aspergillus spp. pose a therapeutic challenge because of their decreased susceptibility to azole drugs. In this review, we outline the microbiological and clinical characteristics of IA due to Aspergillus spp. of section Usti and discuss the therapeutic options.

Highlights • Vermiculite is a new promising carrier for solid-state fermentation of micromycetes. • Aspergillus have higher protease production on vermiculite in comparison with on other carriers. • The mycelium productivity of Aspergillus was higher not only in comparison with SmF, but also in comparison with other carriers under SSF conditions (from 3 to 18 times)., A new method has been developed to increase the productivity of aspergilli - producers of extracellular proteinases based on their cultivation on vermiculite under solid-state fermentation conditions. The productivity of the mycelium Aspergillus ochraceus L-1 and Aspergillus ustus 1 was 3-18 times higher not only in comparison with submerged cultivation, but also in comparison with growth on other carriers studied under solid-state fermentation conditions. Vermiculite can be considered as a new promising carrier for solid-state fermentation of micromycetes.

Background Caffeine, theobromine and theophylline are main purine alkaloid in tea. Theophylline is the downstream metabolite and it remains at a very low level in Camellia sinensis. In our previous study, Aspergillus sydowii could convert caffeine into theophylline in solid-state fermentation of pu-erh tea through N-demethylation. In this study, tea-derived fungi caused theophylline degradation in the solid-state fermentation. The purpose of this study is identify and isolate theophylline-degrading fungi and investigate their application in production of methylxanthines with theophylline as feedstock through microbial conversion. Results Seven tea-derived fungi were collected and identified by ITS, β-tubulin and calmodulin gene sequences, Aspergillus ustus, Aspergillus tamarii, Aspergillus niger and A. sydowii associated with solid-state fermentation of pu-erh tea have shown ability to degrade theophylline in liquid culture. Particularly, A. ustus and A. tamarii could degrade theophylline highly significantly (p A. ustus and A. tamarii, respectively. The data from absolute quantification analysis suggested that 3-methylxanthine and xanthine were the main degraded metabolites in A. ustus and A. tamarii, respectively. 129.48 ± 5.81 mg/L of 3-methylxanthine and 159.11 ± 10.8 mg/L of xanthine were produced by A. ustus and A. tamarii in 300 mg/L of theophylline liquid medium, respectively. Conclusions For the first time, we confirmed that isolated A. ustus, A. tamarii degrade theophylline through N-demethylation and oxidation. We were able to biologically produce 3-methylxanthine and xanthine efficiently from theophylline through a new microbial synthesis platform with A. ustus and A. tamarii as appropriate starter strains.

The aim of research was to characterize epiphyte micromycetes observed on variable cotton fibers accessions, to estimate the range of fiber destruction and select cotton, which were the most resistant to fungus damage. The accessions of differently colored Upland Cotton varieties (Gossypium hirsutum L.) evaluated: eleven cotton of natural green, twelve – of brown and eleven of conventional white color. Cotton plants have been grown in Sothern Federal District, RF. The fiber samples for the study were placed into a thermostat in sterile Petri dishes on moistened filter paper in order to stimulate the development of mycelium or sporulation of fungi naturally occurred on fibers. Incubation carried out in a thermostat at a +24-28 °C, humidity of 90-100% and exposed for 28 days. The samples examined with a microscope or binocular magnifier. Aspergillus ustus (Bainier), A. fumigatus Fresen., A. niger v. Tiegh., A. flavus Link, Penicillium aurantiogriseum Dierckx, P. notatum Westling, Rhizopus nigricans Ehrenb. and Alternaria alternata (Fuier) Keissler were detected. Compared with exposed white, accession of green and brown colors were significantly resistant to fungus. The mean of destruction (K) of white cotton varied up to 0.95, but colored accessions not exceeded 0.3 (initial destruction of the surface, not affecting internal fiber’s structure).

Background. Aspergillus spp. of section Usti (A. ustus) represent a rare cause of invasive aspergillosis (IA). This multicenter study describes the epidemiology and outcome of A. ustus infections. Methods. Patients with A. ustus isolated from any clinical specimen were retrospectively identified in 22 hospitals from 8 coun- tries. When available, isolates were sent for species identification (BenA/CaM sequencing) and antifungal susceptibility testing. Additional cases were identified by review of the literature. Cases were classified as proven/probable IA or no infection, according to standard international criteria. Results. Clinical report forms were obtained for 90 patients, of whom 27 had proven/probable IA. An additional 45 cases were identified from literature review for a total of 72 cases of proven/probable IA. Hematopoietic cell and solid-organ transplant recipi- ents accounted for 47% and 33% cases, respectively. Only 8% patients were neutropenic at time of diagnosis. Ongoing antimold prophylaxis was present in 47% of cases. Pulmonary IA represented 67% of cases. Primary or secondary extrapulmonary sites of infection were observed in 46% of cases, with skin being affected in 28% of cases. Multiple antifungal drugs were used (consecu- tively or in combination) in 67% of cases. The 24-week mortality rate was 58%. A. calidoustus was the most frequent causal agent. Minimal inhibitory concentrations encompassing 90% isolates (MIC90) were 1, 8, >16, and 4 µg/mL for amphotericin B, voriconazole, posaconazole, and isavuconazole, respectively. Conclusions. Aspergillus ustus IA mainly occurred in nonneutropenic transplant patients and was frequently associated with extrapulmonary sites of infection. Mortality rate was high and optimal antifungal therapy remains to be defined.

Improper disposal and spills of spent engine oil into the environment can result in contamination, which eventually affects humans through the food chain. Mycoremediation is an effective and inexpensive alternative to clean up spent engine oil contamination. In recent work, the potential effectiveness of fungi for degrading spent engine oil was confirmed, with the species identified through molecular identification. Fungi that were able to grow in Bushnell Haas Broth supplied with spent engine oil were identified with the potential to utilize spent engine oil as a carbon source. Six species of fungi namely Penicillium simplicissimum, Aspergillus nidulans, Aspergillus niger, Trichoderma longibrachiatum, Aspergillus ustus, and Aspergillus flavus were successfully identified in this study. Over a course of seven days, P. simplicissimum (21.11 percent) was identified as the most effective fungi in degrading spent engine oil, followed by A. nidulans (17.75 percent), A. niger (15.85 percent), T. longibrachiatum (15.12 percent), A. ustus (15.02 percent), and A. flavus (11.80 percent). As these species of fungi were isolated from the natural environment in Peninsular Malaysia, the potential of using these fungi as mycoremediation of spent engine oil was therefore confirmed.

Seven new ophiobolins (1–5, 12, and 14) along with the 11 known analogues (6–11, 13, 15–18) were isolated from the ethyl acetate extracts of the liquid and solid cultures of the mangrove fungus Aspergillus ustus 094102. The structures including the absolute configurations of the seven new compounds were elucidated by spectroscopic analysis, chemical methods, and quantum ECD calculations. Compounds 4–8 and 11–15 showed cytotoxicities against the G3K, MCF-7, MD-MBA-231, MCF/Adr, A549, and HL-60 human cancer cell lines with the IC50 values ranging from 0.6 to 9.5 μM.

It has been shown that micromycetes Aspergillus ustus 1 and Tolypocladium inflatum k1 secrete proteolytic enzymes that possess high collagenolytic, fibrinolytic, and elastolytic activity. The activity of proteinases hydrolyzing fibrillar proteins, which was determined by the cleavage of azo-collagen, was 122.6 × 10–3EAzc/mL in A. ustus 1 and 69.7 × 10–3EAzc/mL in T. inflatum k1 (EAzc is the amount of azocollagen cleaved in 1 min (μg). The maximum values of activity were observed during submerged cultivation of A. ustus 1 for 4 days and of T. inflatum k1 for 5 days. It has been shown that the maximum of collagenolytic and general proteolytic activity during the cultivation of A. ustus 1 are time-separated, unlike T. inflatum k1, which, presumably, can simplify the procedure for obtaining proteinases active against fibrillar proteins.

Aspergillus spp. cause serious invasive lung infections, and Aspergillus fumigatus is the most commonly encountered clinically significant species. Voriconazole is considered to be the drug of choice for treating A. fumigatus infections; however, rising resistance rates have been reported. We evaluated a matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS)-based method for the differentiation between wild-type and non-wild-type isolates of 20 Aspergillus spp. (including 2 isolates of Aspergillus ustus and 1 of Aspergillus calidoustus that were used as controls due their intrinsic low azole susceptibility with respect to the in vitro response to voriconazole). At 30 and 48 h of incubation, there was complete agreement between Cyp51A sequence analysis, broth microdilution, and MALDI-TOF MS classification of isolates as wild type or non-wild type. In this proof-of-concept study, we demonstrated that MALDI-TOF MS can be used to accurately detect A. fumigatus strains with reduced voriconazole susceptibility. However, rather than proving to be a rapid and simple method for antifungal susceptibility testing, this particular MS-based method showed no benefit over conventional testing methods.

Invasive aspergillosis (IA) is the most common invasive fungal infection following a hematopoietic cell transplant, with emerging cryptic species exhibiting resistance to commonly used antifungals such as azoles. These species have been increasingly found after the introduction of anti-mold prophylaxis. We report a case of a 56-year-old female with primary myelofibrosis whose allogeneic hematopoietic cell transplant was complicated by disseminated fungal infection (skin, lung) due to Aspergillus calidoustus, a cryptic specie. Treatment of Aspergillus species remains challenging as these cryptic species are usually resistant to azoles including voriconazole which is the first line of treatment of IA. Infection was successfully treated with surgical excision and combination antifungal therapy based on in vitro susceptibility and synergy testing. Therapy included isavuconazole, a drug that has been shown to be non-inferior to voriconazole in the treatment of invasive mold infections.

Background Aspergillus spp. of section Usti (A. ustus) represent a rare cause of invasive aspergillosis (IA). This multicenter study describes the epidemiology and outcome of A. ustus infections. Methods Patients with A. ustus isolated from any clinical specimen were retrospectively identified in 22 hospitals from 8 countries. When available, isolates were sent for species identification (BenA/CaM sequencing) and antifungal susceptibility testing. Additional cases were identified by review of the literature. Cases were classified as proven/probable IA or no infection, according to standard international criteria. Results Clinical report forms were obtained for 90 patients, of whom 27 had proven/probable IA. An additional 45 cases were identified from literature review for a total of 72 cases of proven/probable IA. Hematopoietic cell and solid-organ transplant recipients accounted for 47% and 33% cases, respectively. Only 8% patients were neutropenic at time of diagnosis. Ongoing antimold prophylaxis was present in 47% of cases. Pulmonary IA represented 67% of cases. Primary or secondary extrapulmonary sites of infection were observed in 46% of cases, with skin being affected in 28% of cases. Multiple antifungal drugs were used (consecutively or in combination) in 67% of cases. The 24-week mortality rate was 58%. A. calidoustus was the most frequent causal agent. Minimal inhibitory concentrations encompassing 90% isolates (MIC90) were 1, 8, >16, and 4 µg/mL for amphotericin B, voriconazole, posaconazole, and isavuconazole, respectively. Conclusions Aspergillus ustus IA mainly occurred in nonneutropenic transplant patients and was frequently associated with extrapulmonary sites of infection. Mortality rate was high and optimal antifungal therapy remains to be defined.

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The contamination of agricultural lands by pesticides, their overuse and toxicity to many non-target organisms cause serious environmental problems. Therefore, the development of bioremediation methods for distinct active ingredients, such as pyrethroids, is primordial. In this work, the biodegradation of gamma-cyhalothrin (GCH) by fungi from the cave Gruta do Catão was investigated. Experiments were conducted with different fungi strains (Aspergillus ustus CBMAI 1894, Talaromyces brunneus CBMAI 1895 and Aspergillus sp. CBMAI 1926) in 2% Malt liquid medium with 300 mg L-1 of GCH in triplicate employing a validated method. All strains biodegraded this insecticide and the most efficient was Aspergillus ustus with a degradation rate of approximately 50% of the initial GCH concentration in 21 days of experiment. We also conducted an experimental design testing three variables: temperature (25-35 degrees), pH (5,5– 8,5) and pesticide concentration (50-550 mg L-1) being the temperature and pesticide concentration were those that most influenced the biodegradation. So, we concluded that cave fungi biodegraded GCH and can be employed in future studies for enzymatic characterization and bioremediation of contaminated environments. A contaminação de áreas agriculturáveis por pesticidas, seu uso intensivo e toxicidade para muitos organismos não alvo causam sérios problemas ambientais. Portanto, o desenvolvimento de metodologias de biorremediação para ingredientes ativos distintos, tais como piretróides, é primordial. Nesse trabalho, a biodegradação de gama cialotrina (GCH) por fungos da Gruta do Catão foi investigada. Os experimentos foram realizados com três diferentes linhagens fúngicas (Aspergillus ustus CBMAI 1894, Talaromyces brunneus CBMAI 1895 e Aspergillus sp. CBMAI 1926) em meio líquido malte 2% com 300 mg L-1 de GCH em experimentos em triplicata utilizando um método validado. Todas as linhagens biodegradaram esse inseticida e a espécie mais eficiente foi Aspergillus ustus com uma taxa de degradação de aproximadamente 50% da concentração inicial de GCH em 21 dias de experimento. Também conduzimos um planejamento experimental testando três variáveis: temperatura (25-35 graus), pH (5,5 - 8,5) e concentração de pesticida (50-550 mg L-1), sendo que a temperatura e a concentração influenciaram no experimento. Concluímos que fungos de caverna biodegradaram GCH e podem ser empregados em estudos futuros para caracterização enzimática e biorremediação de ambientes contaminados. CNPq: 144586/2017-7

Made available in DSpace on 2020-12-10T19:39:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01. Added 1 bitstream(s) on 2021-07-15T14:37:10Z : No. of bitstreams: 1 S0001-37652019000500601.pdf: 975738 bytes, checksum: 4ec2cf4858b8200fba6761772dcd422f (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Isolation and screening of new fungal strains from extreme and understudied environments. such as caves, is a promising approach to find higher yields enzyme producers. Cellulolytic fungal strains isolated from a Brazilian cave were evaluated for their enzymatic production after submerged (SmF) and solid-state fermentation (SSF). After SmF, three strains were selected for their high enzymatic activities: Aspergillus ustus for endoglucanase (4.76 U/mg), Talaromyces bruneus for beta-glucosidase (11.71 U/mg) and Aspergillus sp. (CBMAI 1926) for total cellulase (1.70 U/mg). After SSF, these strains, showed better yields compared to the reference strain Aspergillus niger 3T5B8. Aspergillus sp. (CBMAI 1926) stood out as a new species that expressed activity of total cellulases (0.10 U/mg) and low protein concentration (0.44 mg/mL). In conclusion, these isolated strains have a more efficient and promising cellulolytic enzyme complex that can be used in fermentation and saccharification processes with a lower protein concentration and a higher enzymatic activity than the reference strain. Therefore, beside the new genetic material characterized, our study highlights the benefits of cave extreme environments exploitation to find new potentially valuable strains. Univ Fed Sao Carlos, Dept Ecol & Biol Evolut, Programa Posgrad Ecol & Recursos Nat, Rodovia Washington Luiz,Km 235, BR-13565905 Sao Carlos, SP, Brazil Univ Estado Sao Paulo Julio de Mesquita Filho, Inst Biociencias Rio Clara, Dept Bioquim & Microbiol, Av 24A,1515, BR-13506900 Rio Claro, SP, Brazil Empresa Brasileira Pesquisa Agr, Embrapa Instrumentacao, Rua 15 Novetnbro 1452, BR-13560970 Sao Carlos, SP, Brazil Univ Estado Sao Paulo Julio de Mesquita Filho, Inst Biociencias Rio Clara, Dept Bioquim & Microbiol, Av 24A,1515, BR-13506900 Rio Claro, SP, Brazil FAPESP: FAPESP 2015/24763-9

Two new austalide meroterpenoids, named austalides V and W (1 and 2), were isolated from the fungus Aspergillus ustus VKM F-4692. Their structures were elucidated by extensive spectroscopic analysis and by comparison with related known compounds. The main structural feature of both compounds is a tetrahydrofuranyl ring (G), a structural fragment, first found in austalides. Austalides V (1) and W (2) were able to inhibit the propagation of prostate and bladder cancer cells; this biologic activity is possibly related to the inhibition of a number of key pathways regulating cell growth and migration.

Interaction between indigenous fungal strains in preventing ochratoxin A (OTA) production by Penicillium verrucosum and Penicillium nordicum was studied in 100 mL of Czapek yeast autolysate (CYA) medium in a 250-mL “U” shaped culture vessel in one end for 3 days. At the end of incubation period, test fungi inoculated and incubated at 27 ± 2 °C for another 14 days to study the inhibition of OTA production was estimated by high performance liquid chromatography (HPLC). Total inhibition of OTA production was recorded with Aspergillus fumigatus , Aspergillus flavus , Aspergillus niger , Aspergillus ustus , Fusarium culmorum , Fusarium graminarium , Fusarium proliferatum , Penicillium chrysogenum , Penicillium expansum and Trichoderma viridae . A significant correlation coefficient ( r ) on growth (0.493, P ≤ 0.0003) and OTA production (0.785, P ≤ 0.0001) was observed between the tested Penicillium species and other co-existing fungi. In conclusion, the present investigation revealed that those indigenous fungi are necessary to minimize potential losses to the poultry farmer and toxicological hazards to the consumer as biological control agent in different foods and feeds.

Two new sesterterpenoids, terretonins H ( 1 ) and I ( 2 ), together with two known compounds, strobilactone A ( 3 ) and cerebroside D ( 4 ), were isolated from the lipophilic extract of the marine-derived fungus Aspergillus ustus KMM 4664. The structures of compounds 1 and 2 were determined based on spectroscopic methods and confirmed by X-ray crystallographic analysis of terretonin H. All compounds were tested for cytotoxic and embryotoxic effects using sperm and developing embryos of the sea urchin Strongylocentrotus intermedius and human cancer cells.

In the indoor environment, people are exposed to several fungal species. Evident dampness is associated with increased respiratory symptoms. To examine the immune responses associated with fungal exposure, mice are often exposed to a single species grown on an agar medium. The aim of this study was to develop an inhalation exposure system to be able to examine responses in mice exposed to mixed fungal species aerosolized from fungus-infested building materials. Indoor airborne fungi were sampled and cultivated on gypsum boards. Aerosols were characterized and compared with aerosols in homes. Aerosols containing 10 7 CFU of fungi/m 3 air were generated repeatedly from fungus-infested gypsum boards in a mouse exposure chamber. Aerosols contained Aspergillus nidulans , Aspergillus niger , Aspergillus ustus , Aspergillus versicolor , Chaetomium globosum , Cladosporium herbarum , Penicillium brevicompactum , Penicillium camemberti , Penicillium chrysogenum , Penicillium commune , Penicillium glabrum , Penicillium olsonii , Penicillium rugulosum , Stachybotrys chartarum , and Wallemia sebi . They were all among the most abundant airborne species identified in 28 homes. Nine species from gypsum boards and 11 species in the homes are associated with water damage. Most fungi were present as single spores, but chains and clusters of different species and fragments were also present. The variation in exposure level during the 60 min of aerosol generation was similar to the variation measured in homes. Through aerosolization of fungi from the indoor environment, cultured on gypsum boards, it was possible to generate realistic aerosols in terms of species composition, concentration, and particle sizes. The inhalation-exposure system can be used to study responses to indoor fungi associated with water damage and the importance of fungal species composition.

It was shown that extracellular proteinases produced by the strains of micromycetes A. ochraceus L-1 and A. ustus 1 differ by the activity at various pH as well as by the intensity of the effect on fibrillar proteins. It was revealed that the proteinases of A. ochraceus L-1 demonstrated maximum activity during the growth of the producer in the nitrate-free growth medium (the pH of enzyme reaction was 8.0), whereas those of A. ustus 1 showed maximal activity during the growth of the micromycete in the medium containing sodium nitrate (the pH of enzyme reaction was 6.0). Values of specific fibrinolytic and collagenolytic activities of A. ochraceus L-1 were 2.2 and 1.6 times higher than those of A. ustus 1. At the same time, A. ustus 1 showed very low values of total proteolytic (caseinolytic) activity and had a high ratio of fibrinolytic activity to total proteolytic (caseinolytic) activity (6.92). It makes the strain a promising producer of proteinases, which hydrolyze fibrin and collagen.

The purpose of research is increase of honey productivity bee colonies. The studies were conducted in the early spring in March, before the departure of the insect from the hive after hibernation. Bee colonies of the experimental group treated with ozone for families for 7 consecutive days at 2 hours per day when the ozone concentration of 6 mg . Bee colonies of the control group not treated with ozone. The results showed the presence of different species of microorganisms in the intestinal tract of bees in the control group, the quantitative indicators of the content of the polymorphic bacteria, staphylococci, streptococci, pseudomonas, fungi is quite large. Also, the experiments proved devastating effect of ozonation for bacteria groups, coliforms, pseudomonas, fungi of the genus aspergillus niger in the experimental group. With regard to the fungi penicillium glaucum and aspergillus ustus, ozone caused a slight decrease in their concentration in the intestinal tract of bees. Therefore, the use of ozone as a means of prevention and treatment of infectious diseases of bees can be justified as it creates the most favorable conditions for further development of bee colonies. However, the fact that the number of fungi after ozonation in the intestinal tract of bees increased, speaks about the resistance of these microorganisms to ozone, and therefore, for suppression of fungal flora, ozonation needs to be done either longer or use a higher concentration of ozone.