Chemical constituents of the fermentation broth of the marine-derived fungus Penicillium roqueforti / / Componentes químicos del caldo de fermentación del hongo de origen marino Penicillium roqueforti

Background. The filamentous fungus Penicillium roqueforti is a well-known multifunctional cell factory of high added-value biomolecules. Aims. The objective of this work was to carry out a detailed analysis of the metabolites present in the culture broth of a new marine-derived Penicillium roqueforti strain isolated in the Canary Islands, Spain. Methods. The fungal biomass production was carried out in liquid-state fermentation, and after 10-12 days of incubation at 22-25 °C, the supernatant mycelia was separated by filtration, and the culture broth (12 l) was stored in a refrigerator at 4 °C for a subsequent liquid–liquid extraction with dichloromethane (3×), in accordance with the modified Kupchan method. The volatile and semi-volatile organic compounds were separated by chromatography and analyzed using GC–MS and NMR spectroscopy analyses. Results. Several volatile organic compounds involved in the fatty acid pathway were identified: a terpenoid, a cyclic dipeptide, phthalates, and an alkyl adipate. In addition, three categories of non-volatile compounds (alkanes, fatty acids and 1-alkanols) were identified by spectroscopy. The results show that the fermented broth of this fungal strain has no mycotoxins under the culture conditions applied.Conclusions. It is hoped that this chemo-specific information will offer critical input for improving the biotechnological applications of this filamentous fungus (AU)

Antecedentes. El hongo filamentoso Penicillium roqueforti es una fábrica celular multifuncional de biomoléculas de alto valor añadido. Objetivos. El objetivo de este trabajo fue realizar un estudio minucioso de los metabolitos presentes en el caldo de cultivo de una nueva cepa de Penicillium roqueforti de origen marino aislada en las Islas Canarias, España. Métodos. La producción de biomasa fúngica se llevó a cabo por fermentación en estado líquido. Tras 10-12 días de incubación a 22-25 °C se separó el micelio sobrenadante por filtración y el caldo de cultivo (12 l) se almacenó en un frigorífico a 4 °C hasta su posterior extracción líquido-líquido con diclorometano (×3) de acuerdo con el método modificado de Kupchan. Los compuestos orgánicos volátiles y semi-volátiles se caracterizaron por GC-MS y análisis espectroscópico (NMR). Resultados. Se identificaron varios compuestos orgánicos volátiles implicados en la ruta de los ácidos grasos, un terpeno, un dipéptido cíclico, varios ftalatos y un adipato de alquilo. Además, fueron identificados por espectroscopía tres tipos de compuestos no volátiles: alcanos, ácido grasos y 1-alcanoles. Los resultados mostraron que el caldo fermentado de esta cepa fúngica no presenta micotoxinas en las condiciones de cultivo empleadas. Conclusiones. Es de esperar que esta información quimio-específica aporte datos críticos para el progreso de las aplicaciones biotecnológicas de este hongo filamentoso (AU)

Enhancement of chitosanase production by cell immobilization of Gongronella sp. JG

Chitosanase production of Gongronella sp. JG cells immobilized in calcium alginate gel and polyurethane foam was compared with that of the free cells, there was a 60% increase in the enzyme yield (2429 U/L) compared to the highest yield obtained from free cells (1513 U/L). The optimal immobilization parameters (concentrations of sodium alginate, calcium chloride, bead inoculums, bead diameter, etc) for the enhanced production of chitosanase were determined as: sodium alginate 2% (w/v), 0.1 M calcium chloride, inoculum 10 mL beads to 100 mL production media and 2.7 mm bead diameter. Maximum chitosanase production was achieved with initial pH of 5.5 and temperature of 30 ºC. The alginate beads had well stability, retained 85% ability of enzyme production even after 7 cycles of repeated batch fermentation. These results showed the immobilization technique was a feasible and economical method for chitosansase production by Gongronella sp. JG.

Isolation and cultivation of fungal strains from in vitro cell cultures of two marine sponges (Porifera: Halichondrida and Haplosclerida)

Despite the large number of reports describing sponge-microbe associations, limited knowledge is available about associated fungi and their relationships with the hosts. In this work, specific fungal strains were obtained directly from in vitro sponge cell cultures (primmorphs) and single sponge cells (cytospins) and compared with those obtained from whole tissue preparations. A total of 27 fungal strains were isolated from the marine sponges Hymeniacidon heliophila and Haliclona melana. Fifteen strains, nine from H. heliophila and six from H. melana, were obtained from whole tissue and were considered as possible mesohyl associated or transient fungi. Twelve strains were isolated from in vitro sponge cell cultures (primmorphs) and were, therefore, considered as cell associated. From these, five different strains were obtained from H. heliophila isolated cells, while five were identified from cytospins and two from primmorphs of H. melana. The fungal strains obtained from cell cultures from both sponge species were different, and none of them were detected in the whole tissue preparations of the same species. Nine H. heliophila and seven H. melana strains shows low similarity with the sequences available in public databases and belong to potentially new species. This is the first report of fungi isolated directly from sponge cells, which allowed the observation and selection of specific strains that probably would not be obtained by usual culture dependent techniques.

Contribuição para o conhecimento de Chytridiomycota da Reserva Biológica de Paranapiacaba, Santo André, SP, Brasil / Contribution to the knowledge of the Chytridiomycota from the Reserva Biológica de Paranapiacaba, Santo André, State of São Paulo, Brazil

Estudo da diversidade de quitridiomicetos de solo e água, na "Reserva Biológica de Paranapiacaba", resultou no isolamento de 29 espécies, com três Blastocladiales, 23 Chytridiales, uma Monoblepharidales e duas Spizellomycetales. Dos táxons isolados, 69 por cento são citados pela primeira vez para a Reserva, 14 por cento para o estado de São Paulo, e 7 por cento para o país. É a primeira menção de Rhizophydium condylosum Karling no Brasil.

The diversity study of the chytrids from soil and water from the "Reserva Biológica de Paranapiacaba", yielded 29 species, with three Blastocladiales, 23 Chytridiales, one Monoblepharidales and two Spizellomycetales. From these, 69 percent taxa were cited for the first time for Reserve, 14 percent for São Paulo State and 7 percent for Brazil. Rhizophydium condylosum Karling is being mentioned for the first time in Brazil.