Culture Medium Factorial Design Optimization for Fibrinolytic Enzymes Production by Bionectria sp.

Thrombotic diseases can be clinically treated with fibrinolytic enzymes and many attempts have been made at laboratory level to increase fibrinolytic enzymes production from microbial sources and to reduce the process cost, including culture medium design, optimization of environmental conditions, and over expression with genetically modified strains. In this contribution we present the optimization of culture medium composition and incubation temperature for fibrinolytic enzyme production by Bionectria sp., a selected fungal strain from Las Yungas (Tucumán). Optimization was carried out at Erlenmeyer scale (100-mL working volume) via factorial design methodology. All trials included a common mineral base (%, w/v: NaCl 0.2, KH2PO4 0.05, MgSO4·7H2O 0.05). According to four factorial designs it could be demonstrated the convenience of using soy peptone as N-source, glucose as C-source, and the possibility to eliminate starch, meat peptone and meat extract from original medium composition, whilst 25°C was selected as the optimal incubation temperature. Results showed that culture medium could be successfully optimized by factorial design, achieving a reduction in the production process costs by means of a decrease in culture medium components, the improvement in culture broth rheology, mycelial morphology and mass/energy transfer, and the subsequent two-fold enhancement in productivity.Fil: Arnau, Victor Gonzalo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Rovati, Jose Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Figueroa, L. I. C.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Fariña, Julia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaXLVI Reunión Anual Sociedad Argentina de Investigaciones Bioquímicas y MolecularesPuerto Madryn, Chubut, ArgentinaArgentinaSociedad Argentina de Investigaciones Bioquímicas y Moleculare

Successful fibrinolytic enzymes production by solid state fermentation with a selected Bionectria sp. wild type strain

Fibrin accumulation in blood vessels increases thrombosis risks promoting myocardial infarction and cardiovascular diseases. Bionectria sp. LY 4.1, a wild fungi isolated from Las Yungas Pedemontana rainforest, was recently reported by our group as a novel and promising fibrinolytic enzyme source. In the present study we attempted the production of fibrinolytic enzymes by solid state fermentation (SSF) with Bionectria sp. LY 4.1 particularly looking for low-cost substrates and simplified operational conditions. The different solid substrates evaluated (polyurethane foam - PUF, bagasse, textured soy flour, soy flour pellets and wheat bran), in adequate amounts, were placed in 500 mL-Erlenmeyer flasks and after sterilization, were inoculated with 10 mL of fresh production medium containing 1 mL of a 72 h-old Bionectria sp. culture. The effect of different conditions of moisture (9, 11, 13, 15, 17%), bed height (1, 2, 4, 6, 8 cm) and particle size (0·25; 0·5; 1 and 2 cm2) was studied. Fibrinolytic activity was determined by the fibrin plate technique and protein concentration by the BCA method. The obtained results confirmed the possibility to produce fibrinolytic enzymes by SSF with Bionectria sp. LY 4.1. Furthermore, assays demonstrated the convenience of using PUF as SS under specific operating conditions over the other substrates tested. The findings herein presented open new frontiers for fungal secondary metabolites production by SSF, allowing to offer an interesting alternative for the production of a high added-value pharmacological product.Fil: Rovati, Jose Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Cabral, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Castellanos, Lucia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Fariña, Julia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaXLVII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología MolecularPotrero de los FunesArgentinaSociedad Argentina de Investigación en Bioquímica y Biología Molecula

Successful fibrinolytic enzymes production by solid state fermentation

Fibrin accumulation in blood vessels increases thrombosis risks promoting myocardial infarction and cardiovascular diseases. Bionectria sp. LY 4.1, a wild fungi isolated from Las Yungas Pedemontana rainforest, was recently reported by our group as a novel fibrinolytic enzyme source. In the present study we attempted the production of fibrinolytic enzymes by solid state fermentation (SSF) with Bionectria sp. LY 4.1 looking for low-cost substrates and simplified operational conditions. The different solid substrates (SS) evaluated (polyurethane foam - PUF, bagasse, textured soy flour, soy flour pellets and wheat bran), in adequate amounts were inoculated with fresh production medium containing 72 h-old Bionectria sp. culture. The effect of different conditions of moisture (9, 11, 13, 15, 17%), bed height (1, 2, 4, 6, 8 cm) and particle size(0.25; 0.5; 1 and 2 cm ) was studied. Fibrinolytic activity was determined by the fibrin plate technique and protein concentration by the BCA method. The obtained results confirmed the possibility to produce fibrinolytic enzymes by SSF with Bionectria sp. LY 4.1. Furthermore, assays demonstrated the convenience of using PUF as SS under specific operating conditions over the other substrates tested. The findings herein presented offer an interesting alternative for the production of a high added-value pharmacological product.Fil: Rovati, Jose Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Cabral, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Castellanos, Lucia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaFil: Fariña, Julia Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; ArgentinaXLVII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología MolecularSan LuisArgentinaSociedad Argentina de Investigación Bioquímica y Biología Molecula

Polyphenolic substrates and dyes degradation by yeasts from 25 de Mayo/King George Island (Antarctica)

Antarctica offers a range of extreme climatic conditions, such as low temperatures, high solar radiation and low nutrient availability, and constitutes one of the harshest environments on Earth. Despite that, it has been successfully colonized by 'cold-loving' fungi, which play a key role in decomposition cycles in cold ecosystems. However, knowledge about the ecological role of yeasts in nutrient or organic matter recycling/mineralization remains highly fragmentary. The aim of this work was to study the yeast microbiota in samples collected on 25 de Mayo/King George Island regarding the scope of their ability to degrade polyphenolic substrates such as lignin and azo dyes. Sixty-one yeast isolates were obtained from 37 samples, including soil, rocks, wood and bones. Molecular analyses based on rDNA sequences revealed that 35 yeasts could be identified at the species level and could be classified in the genera Leucosporidiella, Rhodotorula, Cryptococcus, Bullera and Candida. Cryptococcus victoriae was by far the most ubiquitous species. In total, 33% of the yeast isolates examined showed significant activity for dye decolorization, 25% for laccase activity and 38% for ligninolytic activity. Eleven yeasts did not show positive activity in any of the assays performed and no isolates showed positive activity across all tested substrates. A high diversity of yeasts were isolated in this work, possibly including undescribed species and conspicuous Antarctic yeasts, most of them belonging to oligotrophic, slow-growing and metabolically diverse basidiomycetous genera.Fil: Rovati, Jose Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos (i); Argentina;Fil: Pajot, Hipólito Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos (i); Argentina;Fil: Ruberto, Lucas Adolfo Mauro. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología; Argentina;Fil: Mac Cormack, Walter Patricio. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; Argentina;Fil: Figueroa, Lucía I. C.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos (i); Argentina