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Geothermal areas are the niches of a rich microbial diversity that is not only part of the intangible patrimony of a country but also the source of many microbial species with potential biotechnological applications. Particularly, microbial species in geothermal areas in Argentina have been scarcely explored regarding their possible biotechnological uses. The purpose of this work was to explore the proteolytic and keratinolytic enzymatic potential of microorganisms that inhabit in the Domuyo geothermal area in the Neuquén Province. To this end, we did enrichment cultures from two high-temperature natural samples in mineral media only supplemented with whole chicken feathers. After the isolation and the phylogenetic and morphologic characterization of different colonies, we obtained a collection of Bacillus cytotoxicus isolates, a species with no previous report of keratinolytic activity and only reported in rehydrated meals connected with food poisoning outbreaks. Its natural habitat has been unknown up to now. We characterized the proteolytic and keratinolytic capacities of the B. cytotoxicus isolates in different conditions, which proved to be remarkably high compared with those of other similar species. Thus, our work represents the first report of the isolation as well as the keratinolytic capacity characterization of strains of B. cytotixicus obtained from a natural environment., Centro de Investigación y Desarrollo en Fermentaciones Industriales

La economía colombiana viene creciendo en muchos sectores y uno de los que más se ha reactivado es la construcción, especialmente de edificios y viviendas en las grandes ciudades como Bogotá, Cali, Medellín y Barranquilla. además, las grandes obras de infraestructura vial como carreteras, viaductos, puentes, túneles y obras de geotecnia, vienen utilizando grandes cantidades de materiales de construcción. Se presenta el diseño de una planta de extracción y venta de agregado para construcción ubicada en el corregimiento de Arroyo de Piedra, Atlántico (Colombia), a 60 km de Barranquilla, cuya capacidad de producción total de agregado es de 24.000 metros cúbicos por mes. La planta tiene como objetivo suplir un porcentaje de la demanda de material en el mercado de construcción de Barranquilla y el norte del departamento del Atlántico. Para el diseño de esta planta se tuvieron en cuenta normas y códigos para la extracción y procesamiento de agregados. Se analizaron detenidamente los procesos y subprocesos de la planta para realizar una acertada elección de equipamiento. Y se diseñó detalladamente la banda transportadora principal de la planta bajo los manuales Bridgestone. The Colombian economy has been growing in many sectors and one of the most reactivated is construction, especially of buildings and houses in large cities such as Bogotá, Cali, Medellín and Barranquilla. In addition, large road infrastructure works such as roads, viaducts, bridges, tunnels and geotechnical works have been using large amounts of construction materials. The design of an aggregate extraction and sale plant for construction is presented, located in the corregimiento of Arroyo de Piedra, Atlántico (Colombia), 60 km from Barranquilla, whose total aggregate production capacity is 24,000 cubic meters per month. The plant aims to supply a percentage of the demand for material in the construction market of Barranquilla and the north of the department of Atlántico. For the design of this plant, rules and codes for the extraction and processing of aggregates were taken into account. The processes and subprocesses of the plant were carefully analyzed to make an accurate choice of equipment. And the main conveyor belt of the plant was designed in detail under the Bridgestone manuals.

Aspergillus niger biomass, an industrial by-product of citric acid fermentation is an emergent source of glycoderivatives with applications in biofuel, cosmetics, feed, energy, food, medicine, and nanotechnology. In this study, the effect of purified neutral protease for deprotenization of fungal biomass studied at various levels (0, 5, 10, 20 and 40 U/100 mg of biomass) and the saccharification of fungal biomass was evaluated with amylolytic enzymes and chitosanases. The efficiency of deproteinization of fungal biomass was based on the enzyme concentration and contact time. Protease at a concentration of 20 U/100 mg of dry biomass and with a contact time of 8 h achieved 30% final deproteinization. No effect on saccharification of A. niger biomass was observed by treatment with purified amylolytic enzymes. Meanwhile, the endo- and exo-chitosanases treatment yielded 54 g of g reducing sugars (equivalent to amino sugars)/ kg of fungal biomass, which can be employed for tailor-made carbohydrate production. Fil: Perez Cruz, Cecilia. Universidad Autónoma de Coahuila; México Fil: Cano Gonzalez, Carlos N.. Universidad Autónoma de Coahuila; México Fil: Fuentes, Jose. Universidad Autónoma de Coahuila; México Fil: Balagurusamy, Nagamani. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Vita, Carolina Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Hours, Roque Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Aguilar, Cristobal N.. Universidad Autónoma de Coahuilla; México Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Contreras Esquivel, Juan C.. Universidad Autónoma de Coahuilla; México

Leaf cutter ants are one of the most important pests to agriculture and forestry in the Neotropics. One environmentally friendly solution to control them is biological control with the use of conidia of some fungi, such as Beauveria bassiana. We tested several isolates of this fungus (obtained from leaf cutter ants) by inoculating ants from seven colonies of Acromyrmex lundii with 5 × 106 conidia ml−1 suspensions. All the isolates proved to be good controllers, but isolate 5 (B5) was found to be the best performing option, because it caused the death of 92.3% of the ants in 4–5 days. Isolate 7 (B7) was the second-best option and used for comparative purposes. Further assays were performed with the isolate B5 to optimise yield production. Three types of rice (brown, parboiled and white rice), soybean hull and yerba mate (Ilex paraguariensis leaves) were tested as substrates, and following that, different conditions of solid-state fermentation (SSF) were changed so as to select the most productive combination. Among treatments, we selected brown rice as substrate and a temperature of 28°C and 75–85% relative humidity in the SSF chamber because this allowed a production of 6.4 × 108 and 1.2 × 109 conidia of B5 and B7 per gram of final substrate with a 98% and 97.5% of germination, in 10 days, respectively. The differences in yield obtained under the same conditions for both isolates show that yield is isolate-dependent. A description of the customised design of the SSF chamber used is included. Fil: Goffré, Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Hormigas; Argentina Fil: Cavallo, Ema Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Hormigas; Argentina Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Folgarait, Patricia Julia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Hormigas; Argentina

Exoinulinases—enzymes extensively studied in recent decades because of their industrial applications—need to be produced in suitable quantities in order to meet production demands. We describe here the production of an acid-stable recombinant inulinase from Aspergillus kawachii in the Pichia pastoris system and the recombinant enzyme's biochemical characteristics and potential application to industrial processes. After an appropriate cloning strategy, this genetically engineered inulinase was successfully overproduced in fed-batch fermentations, reaching up to 840 U/ml after a 72-h cultivation. The protein, purified to homogeneity by chromatographic techniques, was obtained at a 42% yield. The following biochemical characteristics were determined: the enzyme had an optimal pH of 3, was stable for at least 3 h at 55 °C, and was inhibited in catalytic activity almost completely by Hg+2. The respective Km and Vmax for the recombinant inulinase with inulin as substrate were 1.35 mM and 2673 μmol/min/mg. The recombinant enzyme is an exoinulinase but also possesses synthetic activity (i. e., fructosyl transferase). The high level of production of this recombinant plus its relevant biochemical properties would argue that the process presented here is a possible recourse for industrial applications in carbohydrate processing. Fil: Chesini, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Plaza Cazón, Josefina del Carmen. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Baruque, Diego Jorge. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Vita, Carolina Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Ghiringhelli, Pablo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina Fil: Rojas, Natalia Lorena. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Thermophilic aerobic bacteria were isolated from two geothermal areas in Neuquén province using two different enrichment methods and a total of 30 isolates were obtained. From chicken feather enrichment cultures, strains affiliated to Bacillus cytotoxicus and Bacillus licheniformis were isolated and all of them demonstrated the capability to degrade completely chicken feather. A preliminary research on biotechnological enzymes’ potential demonstrated that all the isolates displayed at least one of the extracellular hydrolytic enzymes tested. Most of the isolates showed protease, inulinase and/or pectinase activities, while cellulase and xylanase activities were less common. In light of these findings, geothermal areas of Argentina may be considered as a potential source of thermophilic bacteria able to produce many industrially relevant enzymes., Facultad de Ciencias Exactas, Centro de Investigación y Desarrollo en Fermentaciones Industriales

New enzymes isolated from the biodiversity of native wine ecosystems could contribute to increase the varietal character of regional wines. This study reports on the production and characterization of Issatchenkia terricola, beta-glucosidase and its potential to release red-wine aromatic compounds. The enzyme, a monomer of 48 kDa with an isoelectric point of 3.5 is tolerant to glucose and ethanol, properties compatible with enological use. Although fed-batch is usually the most suitable system for enzyme production in submerged culture, in this case the yield was practically the same as in batch culture. Enzyme productivity was increased 2-fold in synthetic medium with glucose with respect to the YPG and 3 to 8-fold with respect to other media assayed. After enzymatic treatment, GC-MS analysis of the released aglycones demonstrated significant increases in the concentration of phenols (83%) and norisoprenoids (65%). According to the judges of the sensory panel, the treatment resulted in a wine with dried fruits and raisins notes, as compared to the control, which was found more sweet and fruity. This, together with the lack of activity on anthocyanin glycosides, highlights the potential of this enzyme in enology, since its high selectivity allowed the development of aroma without compromising wine color., Centro de Investigación y Desarrollo en Fermentaciones Industriales

An endo-polygalacturonase secreted by Aspergillus sojae was characterized after being purified to homogeneity from submerged cultures with orange peel as the sole carbon source by gel filtration and ion-exchange chromatographies. According to SDS-PAGE and analytical isoelectric focusing analyses, the enzyme presents a molecular weight of 47 kDa and pI value of 4.2. This enzyme exhibits considerable stability under highly acidic to neutral conditions (pH 1.5-6.5) and presents a half-life of 2 h at 50°C. Besides its activity towards pectin and polygalacturonic acid, the enzyme displays pectin-releasing activity, acting best in a pH range of 3.3-5.0. Thin-layer chromatographic analysis revealed that tri-galacturonate is the main enzymatic end product of polygalacturonic acid hydrolysis, indicating that it is an endo-polygalacturonase. The enzyme exhibits Michaelis-Menten kinetics, with KM and VMAX values of 0.134 mg/mL and 9.6 μmol/mg/min, respectively, and remained stable and active in the presence of SO2, ethanol, and various cations assayed except Hg2+. Fil: Fratebianchi de la Parra, Dante. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Cavello, Ivana Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina

The glycoprotein (G-protein) of rabies virus is responsible for viral attachment to the host cell surface and induces virus neutralization antibodies. In the present study, the G-protein gene of rabies virus CVS strain was cloned, sequenced and expressed in the yeast, Pichia pastoris, as a secreted protein, using a simplified DO-stat control feeding strategy. This strategy involves the addition of methanol when the dissolved oxygen (DO) level rises above the setpoint avoiding methanol accumulation and oxygen limitation. The G-protein expression was evaluated by SDS-PAGE, ELISA, and western blot assays. Like native G-protein, the recombinant G-protein was found reactive when it was challenged against specific antibodies. The data indicate that the recombinant G-protein can be easily expressed and isolated, and may be useful as a safe source in the production of diagnostic kits and subunit vaccines to prevent rabies., Facultad de Ciencias Veterinarias, Centro de Investigación y Desarrollo en Fermentaciones Industriales

One hundred and three yeasts isolated from soil samples from King George Island and Tierra del Fuego province were screened in relation with their capability to produce pectinolytic enzymes. Although all the yeasts showed well-developed colonies at 20 °C, only eight showed a clear halo around the colony, indicative of pectin degradation. A secondary screening demonstrated that only four yeasts were capable to produce pectinases at low temperatures (8 °C). It could be seen that the selected yeasts were able to grow and produce high levels of polygalacturonase activity when submerged fermentations were performed using pectin-containing fruit wastes as substrates. None of the strains produced neither lyase nor rhamnogalacturonan hydrolase activities. Regarding pectin esterase activity, it was only produced in lower amounts by G. pullulans 8E (0.022 U ml−1). A TLC analysis of the substrate cleavage pattern of the pectinolytic systems was consistent with an endo-type activity. The clarification of apple juice was only accomplished by G. pullulans pectinolytic system, with a clarification of 80% (%T650) using 4 U/ml of enzyme at 20 °C. As far as we concern this work describes for the first time the production of pectinases by the cold-adapted yeasts species Cystofilobasidium infirmominiatum, Cryptococcus adeliensis and G. pullulans. Fil: Cavello, Ivana Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Albanesi, Agustin Pedro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Fratebianchi de la Parra, Dante. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Garmedia, Gabriela. Universidad de la República; Uruguay Fil: Vero, Silvana. Universidad de la República; Uruguay Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina

BACKGROUND: The koji mold Aspergillus sojae, an industrially important microorganism, can produce high levels of pectinases utilizing agro-industrial wastes. This study introduces apricot and peach pomace, two agro-industrial wastes barely considered as raw material for the generation of value-added products, and focuses on its utilization together with orange peel for polygalacturonase production in submerged cultures using A. sojae. RESULTS: A Doehlert response surface methodology design conducted in shake flasks and applied individually with these three by-products led to 60–80 U mL−1 polygalacturonase activity. In bioreactor studies performed with a mixture of apricot pomace and orange peel, by fixing stirrer speed to 600 rpm and cascading airflow to the dissolved oxygen tension up to 1.7 vvm, oxygen limitation problems were overcome and polygalacturonase activity values of 380 U mL−1 were achieved. CONCLUSION: A simple and efficient strategy to minimize oxygen limitation with the lowest possible shear stress is provided for stirred-tank bioreactors working with highly viscous broths, so as to ultimately enhance microbial enzyme production. The polygalacturonase activity yields obtained in our study are among the highest reported in the literature., Centro de Investigación y Desarrollo en Fermentaciones Industriales

Various microbial groups are well known to produce a range of extracellular enzymes and other secondary metabolites. However, the occurrence and importance of investment in such activities have received relatively limited attention in studies of Antarctic soil microbiota. Sixty-one yeasts strains were isolated from King George Island, Antarctica which were characterized physiologically and identified at the molecular level using the D1/D2 region of rDNA. Fifty-eight yeasts (belonging to the genera Cryptococcus, Leucosporidiella, Rhodotorula, Guehomyces, Candida, Metschnikowia and Debaryomyces) were screened for extracellular amylolytic, proteolytic, esterasic, pectinolytic, inulolytic xylanolytic and cellulolytic activities at low and moderate temperatures. Esterase activity was the most common enzymatic activity expressed by the yeast isolates regardless the assay temperature and inulinase was the second most common enzymatic activity. No cellulolytic activity was detected. One yeast identified as Guehomyces pullulans (8E) showed significant activity across six of seven enzymes types tested. Twenty-eight yeast isolates were classified as oleaginous, being the isolate 8E the strain that accumulated the highest levels of saponifiable lipids (42 %). Fil: Martinez, A.. Universidad de la República. Facultad de Química; Uruguay Fil: Cavello, Ivana Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Garmendia, G.. Universidad de la República. Facultad de Química; Uruguay Fil: Rufo, C.. Universidad de la República. Facultad de Química; Uruguay Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Vero, S.. Universidad de la República. Facultad de Química; Uruguay

The potential of important agro-industrial wastes, apple pomace (AP) and orange peel (OP) as C sources, was investigated in the maximization of polygalacturonase (PG), an industrially significant enzyme, using an industrially important microorganism Aspergillus sojae. Factors such as various hydrolysis forms of the C sources (hydrolysed-AP, non-hydrolysed-AP, hydrolysed-AP + OP, non-hydrolysed-AP + OP) and N sources (ammonium sulphate and urea), and incubation time (4, 6, and 8 days) were screened. It was observed that maximum PG activity was achieved at a combination of non-hydrolysed-AP + OP and ammonium sulphate with eight days of incubation. For the pre-optimization study, ammonium sulphate concentration and the mixing ratios of AP + OP at different total C concentrations (9, 15, 21 g l−1) were evaluated. The optimum conditions for the maximum PG production (144.96 U ml−1) was found as 21 g l−1 total carbohydrate concentration totally coming from OP at 15 g l−1 ammonium sulphate concentration. On the other hand, 3:1 mixing ratio of OP + AP at 11.50 g l−1 ammonium sulphate concentration also resulted in a considerable PG activity (115.73 U ml−1). These results demonstrated that AP can be evaluated as an additional C source to OP for PG production, which in turn both can be alternative solutions for the elimination of the waste accumulation in the food industry with economical returns., Centro de Investigación y Desarrollo en Fermentaciones Industriales

The application of cellulases in saccharification processes is restricted by its production cost. Consequently, new fungal strains able to elaborate higher cellulases titers and with special activity profiles are required to make the process economical. The aim of this investigation was to find a promising wild-type Trichoderma strain for cellulases production. The Trichoderma reesei strain 938 (CBS 836.91) was selected among twenty strains on the basis of cellulase-agar-plate screening. Evaluation of the selected strain on six solid substrates indicated the highest activities to be obtained from wheat bran. Statistical analyses of the experimental design indicated a significant effect of pH and moisture on the generation of endoglucanase (EGA) and filter-paper (FPA) activity. Furthermore, a central-composite design-based optimization revealed that pH values between 6.4 and 6.6 and moisture from 74 to 94 % were optimal for cellulases production. Under these conditions, 8?10 IU gds-1 of FPA and 15.6?17.8 IU gds-1 of EGA were obtained. In addition, cultivation in a rotating-drum reactor under optimal conditions gave 8.2 IU gds-1 FPA and 13.5 IU gds-1 EGA. Biochemical characterization of T. reesei 938 cellulases indicated a substantially higher resistance to 4 mM Fe?2 and a slightly greater tolerance to alkaline pH in comparison to Celluclast. These results suggest that T. reesei 938 could be a promising candidate for improved cellulases production through direct-evolution strategies. Fil: Ortiz, Gastón Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Universitat Bremen. School Of Enigineerring And Science Jacobs; Alemania Fil: Guitart, María Eugenia. Universitat Bremen. School Of Enigineerring And Science Jacobs; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - la Plata. Centro de Investigación y Desarrollo En Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Cs.exactas. Centro de Investigación y Desarrollo En Fermentaciones Industriales; Argentina Fil: Alberto, Edgardo Omar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Fernandez Lahore, Marcelo. Universitat Bremen. School Of Enigineerring And Science Jacobs; Alemania Fil: Blasco, Martín. Instituto Nacional de Tecnología Industrial. Centro de Investigación y Desarrollo en Biotecnología Industrial; Argentina

Purpureocillium lilacinum (Thom) Samsom is one of the most studied fungi in the control of plant parasitic nematodes. However, there is not specific information on its ability to inhibit some pathogenic bacteria, fungi, or yeast. This work reports the production of several antifungal hydrolytic enzymes by a strain of P. lilacinum when it is grown in a medium containing hair waste. The growth of several plant-pathogenic fungi, Alternaria alternata, Aspergillus niger, and Fusarium culmorum, was considerably affected by the presence of P. lilacinum’s supernatant. Besides antifungal activity, P. lilacinum demonstrates the capability to produce indoleacetic acid and ammonia during time cultivation on hair waste medium. Plant growth-promoting activity by cell-free supernatant was evidenced through the increase of the percentage of tomato seed germination from 71 to 85% after 48 hours. A 21-day plant growth assay using tomato plants indicates that crude supernatant promotes the growth of the plants similar to a reference fertilizer ( > 0.05). These results suggest that both strain and the supernatant may have potential to be considered as a potent biocontrol agent with multiple plant growth-promoting properties. To our knowledge, this is the first report on the antifungal, IAA production and tomato growth enhancing compounds produced by P. lilacinum LPSC #876., Centro de Investigación y Desarrollo en Fermentaciones Industriales

Keratinase from Purpureocillium lilacinum LPSC # 876 was immobilized on chitosan beads using two different cross-linking agents: glutaraldehyde and genipin. For its immobilization certain parameters were optimized such as cross-linker concentration, activation time and activation temperature. Under optimum conditions, enzyme immobilization resulted to be 96 and 92.8% for glutaraldehyde and genipin, respectively, with an activity recovery reaching up to 81% when genipin was used. The immobilized keratinase showed better thermal and pH stabilities compared to the soluble form, retaining more than 85% of its activity at pH 11 and 74% at 50 °C after 1 h of incubation. The residual activity of immobilized keratinase remained more than 60% of its initial value after five hydrolytic cycles. The results in this study support that glutaraldehyde could be replaced by genipin as an alternative cross-linking eco-friendly agent for enzyme immobilization. Fil: Cavello, Ivana Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - la Plata. Centro de Investigación y Desarrollo En Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Cs.exactas. Centro de Investigación y Desarrollo En Fermentaciones Industriales; Argentina Fil: Contreras Esquivel, Juan Carlos. Universidad Autónoma de Coahuila; México Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - la Plata. Centro de Investigación y Desarrollo En Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Cs.exactas. Centro de Investigación y Desarrollo En Fermentaciones Industriales; Argentina

El residuo pelo es conocido como un residuo sólido que es generado luego de un proceso de pelambre con recupero de pelo en la tecnología del cuero, y es producido en grandes cantidades en la provincia de Buenos Aires, Argentina. El método actual de tratamiento de este residuo es la disposición del mismo, lo que es problemático para el medio ambiente y costoso para las curtiembres. Su biodegradación por microorganismos queratinolíticos representa una alternativa para el tratamiento del mismo. En el presente trabajo se presenta la optimización de la producción de queratinasas por Purpureocillium lilacinum LPS # 876. En primer lugar, se utilizó el método de un factor a la vez, para investigar el efecto de las fuentes de nitrógeno, la concentración de iones metálicos en el medio mineral basal, así como, el pH inicial y la temperatura en la producción de la enzima. La composición del medio de cultivo para la producción de queratinasas por P. lilacinum se optimizó por el uso de dos métodos estadísticos: el diseño Plackett-Burman y el diseño central compuesto -combinado con metodologia de superficies de respuestautilizada para optimizar las tres variables estadísticamente representativas: pH inicial, glucosa y CaCl2 . Se determinaron las condiciones óptimas como 7.10 g l-1 de glucosa; 0.0065 mg l -1 de CaCl2 y pH inicial de 5,60; en esta condición, el rendimiento máximo predecible en actividad queratinolítica fue 26.7 U ml-1. La validación del modelo mostró que el modelo propuesto describe adecuadamente la influencia de la concentración de glucosa y el calcio y el pH inicial en la producción de enzimas queratinoliticas. La producción de queratinasas por P. lilacinum es un campo poco explotado que podría representar una nueva y prometedora aplicación biotecnológica para este microorganismo Hair waste is recognized as a solid waste generated after a hair-saving unhairing process in leather technology, and it is produced in large amounts in Buenos Aires province, Argentine. The present method for this solid waste is disposal, which is environmentally troublesome and costly for the tanneries. Biodegradation of hair waste by keratinolytic microorganisms represents an alternative for bioconversion of keratin waste. This paper reports the optimization of keratinases production by Purpureocillium lilacinum LPS # 876. Firstly, one factor-at-a-time method was used to investigate the effect of nitrogen-sources, metal ions concentration in the basal mineral medium, as well as, initial pH and temperature on enzyme production. Medium composition for the keratinases production by P. lilacinum was optimized using two statistical methods: Plackett-Burman design and central composite design -combined with response surface methodology- used to optimize three significant variables: initial pH, glucose and CaCl2 . The optimal conditions were determined as 7.10 g l-1 of glucose; 0.0065 mg l -1 of CaCl2 and initial pH of 5.60; at this condition the maximum keratinase yield predicted was 26.7 U ml-1. The model validation showed that it described adequately the influence of glucose and calcium concentration and initial pH on enzyme production. Production of keratinases by P. lilacinum is a less-exploited field that might represent a novel and promising biotechnological application for this microorganism. Fil: Cavello, Ivana Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Centro de Investigación y Desarrollo En Fermentaciones Industriales (i); Argentina Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Centro de Investigación y Desarrollo En Fermentaciones Industriales (i); Argentina

Background: Inulinases have been extracted and characterized from inulin-storing tissues; however, production of microbial inulinases have recently draw much attention as they offer several industrial advantages. Many microorganisms, including filamentous fungi, yeast and bacteria have been claimed as inulinase producers. These hydrolases are usually inducible and their exo-acting forms may hydrolyze fructose polymers (inulin) and oligosaccharides such as sucrose and raffinose. Fungal inulinase extracts are often produced as stable mixture of highly active fructanhydrolases. From a practical prospective, the best known inulinases to date are those produced by species of Penicillium, Aspergillus and Kluyveromyces. Results: The production of extracellular inulinase by A. kawachii in liquid cultures, using either inulin or yacon derived materials as CES as well as inulinase inducers, is reported. In addition, a partial characterization of the enzyme activity is included. Conclusions: Yacon derived products, particularly yacon juice, added to the culture medium proved to be a good CES for fungal growth as well as an inducer of enzyme synthesis. Partial characterization of the enzyme revealed that it is quite stable in a wide range of pH and temperature. In addition, characterization of the reaction products revealed that this enzyme corresponds to an exo-type. These facts are promising considering its potential application in inulin hydrolysis for the production of high fructose syrups., Centro de Investigación y Desarrollo en Fermentaciones Industriales

A keratinolytic serine protease secreted by Purpureocillium lilacinum (formerly Paecilomyces lilacinus) upon culture in a basal medium containing 1% (w/v) hair waste as carbon and nitrogen source was purified and characterized. After purification the keratinase was resolved by SDS-PAGE as a homogeneus protein band of molecular mass 37.0 kDa. The extracellular keratinase of P. lilacinum was characterized by its appreciable stability over a broad pH range (from 4.0 to 9.0), and up to 65 °C, along with its strong inhibition by phenylmethylsulphonyl fluoride among the protease inhibitors tested (98.2% of inhibition), thus suggesting its nature as a serine protease. The enzyme was active and stable in the presence of organic solvents such as dimethylsulfoxide, methanol, and isopropanol; certain surfactants such as Triton X-100, sodium dodecylsulfate, and Tween 85; and bleaching agents such as hydrogen peroxide. These biochemical characteristics suggest the potential use of this enzyme in numerous industrial applications. Fil: Cavello, Ivana Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Centro de Investigación y Desarrollo En Fermentaciones Industriales (i); Argentina Fil: Hours, Roque Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Centro de Investigación y Desarrollo En Fermentaciones Industriales (i); Argentina Fil: Rojas, Natalia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Centro de Investigación y Desarrollo En Fermentaciones Industriales (i); Argentina Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico la Plata. Centro de Investigación y Desarrollo En Fermentaciones Industriales (i); Argentina

A keratinase isolated from Paecilomyces lilacinus (LPS #876) was tested against proteins present in the skin but the high enzyme activity was detected on collagen. Keratinase was physically immobilized onto PVA–pectin cryogels and enzyme release was 20.8 ± 2.1%, 63.8 ± 0.2%, 41.5 ± 3.5% and 26.0 ± 3.5% in cryogels containing pectins with esterification degrees (DE) 33.0%, 55.0%, 62.7% and 71.7% respectively at 37 ºC after 3 h incubation. In presence of 0.75 M NaCl, the percentage of enzyme release changed to: 57.5 ± 1.5, 65.8 ± 3.8, 57.3 ± 0.2 and 34.0 ± 4.0 for the four pectins respectively. In-vitro studies of enrofloxacin release from PVA–pectin cryogels at pH close to the human skin (pH = 5.5) showed 15.0% free antibiotic following first order kinetic at 37 ºC after 5 h incubation. However, in the presence of keratinase only 6.9% of enrofloxacin was released under the same experimental conditions., Centro de Investigación y Desarrollo en Fermentaciones Industriales

Paecilomyces lilacinus (Thom) Samson LPS 876, a locally isolated fungal strain, was grown on minimal mineral medium containing “hair waste,” a residue from the hair-saving unhairing process, and produced a protease with keratinolytic activity. This enzyme was biochemically characterized. The optimum reaction conditions, determined with a response surface methodology, were 60°C and pH 6.0. It was remarkably stable in a wide range of pHs and temperatures. Addition of Ca2+, Mg2+, or sorbitol was found to be effective in increasing thermal stability of the protease. PMSF and Hg2+ inhibited the proteolytic activity indicating the presence of a thiol-dependent serine protease. It showed high stability toward surfactants, bleaching agents, and solvents. It was also compatible with commercial detergents (7 mg/mL) such as Ariel, Skip, Drive, and Ace, retaining more than 70% of its proteolytic activity in all detergents after 1 h of incubation at 40°C. Wash performance analysis revealed that this protease could effectively remove blood stains. From these properties, this enzyme may be considered as a potential candidate for future use in biotechnological processes, as well as in the formulation of laundry detergents., Facultad de Ciencias Exactas, Centro de Investigación y Desarrollo en Fermentaciones Industriales

Paecilomyces lilacinus (LPS 876) efficiently degraded keratin in chicken feather during submerged cultivation producing extracellular proteases. Characterization of crude protease activity was done including its compatibility in commercial detergents. Optimum pH and temperature were 10.0 and 60 °C, respectively. Protease activity was enhanced by Ca2+ but was strongly inhibited by PMSF and by Hg2+ suggesting the presence of thiol-dependent serine proteases. The crude protease showed extreme stability toward non-ionic (Tween 20, Tween 85, and Triton X-100) and anionic (SDS) surfactants, and relative stability toward oxidizing agent (H2O2 and sodium perborate). In addition, it showed excellent stability and compatibility with various solid and liquid commercial detergents from 30 to 50 °C. The enzyme preparation retained more than 95% of its initial activity with solid detergents (Ariel™ and Drive™) and 97% of its original activity with a liquid detergent (Ace™) after pre-incubation at 40 °C. The protective effect of polyols (propylene glycol, PEG 4000, and glycerol) on the heat inactivation was also examined and the best results were obtained with glycerol from 50 to 60 °C. Considering its promising properties, P. lilacinus enzymatic preparation may be considered as a candidate for use in biotechnological processes (i.e., as detergent additive) and in the processing of keratinous wastes. Fil: Cavello, Ivana Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Cavalitto, Sebastian Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina Fil: Hours, Roque Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina

ATCC 42397 produces a protopectinase (PPase-SE) with polygalacturonase (PGase) activity. The microorganism was aerobically cultivated in synthetic media. Glucose, fructose and xylose yielded the highest enzyme levels (10–11 PGase units ml−1). Galacturonic acid repressed enzyme production and no growth was obtained with disaccharides and pectin. Specific enzyme activity obtained in an O2-limited culture was similar to that found in nonlimited ones. A growth yield (Y; x/s) of 0.49 g of cell dry weight per gram of glucose consumed was obtained in a typical batch bioreactor culture. Enzyme production was growth associated, and no major products other than biomass and CO2 were detected. The volumetric enzyme activity reached a maximum around D=0.3–0.4 h−1 in glucose-limited continuous cultures. However, it varied strongly (together with microorganism morphology) even after retention times ≥8 at any D tested (0.035–0.44 h−1) though the rest of the culture variables remained fairly constant. No correlation between morphology and enzyme activity could be obtained. Enzyme production was poor in urea- and vitamin-limited continuous cultures. In all cases, biomass and CO2 accounted for ≅100% of carbon recovery though Y; x/s values were different., Centro de Investigación y Desarrollo en Fermentaciones Industriales

Solid-state cultures of a pectinase-producing fungus (Aspergillus foetidus NRRL 341) were performed under different acidic conditions. Glass bottles containing 5 g of wheat bran and 7.5 mL of 0.2, 0.3, 0.4, or 0.5 N HCl were autoclaved (15 min, 121°C), inoculated with a spore suspension appropriately diluted to achieve an initial concentration of 4 × 10⁴ spores per gram of wet substrate (with a 60 % humidity, on wet basis) and incubated at 30°C. Time course of pH and of different pectinase activities were determined in culture extracts. Total pectinase activity (TPA), expressed in terms of viscosimetric units per gram of wet substrate (VU.g⁻¹), was affected by the initial culture acidity. The higher the HCl concentration used, the higher the TPA achieved, but after longer cultivation times. On the other hand, when 0.5 HCl was used, no fungal growth was observed. Nevertheless, enzyme productivity increased with culture acidity. When 0.4 HCl was used, TPA reached its maximum after 36 h of cultivation (2,535 VU.g⁻¹). With 0.2 and 0.3 N HCl, TPA was the highest at 24 h (733 VU.g⁻¹) and at 30 h (1,860 VU.g⁻¹) respectively. The composition of the pectinase pool was also affected by culture acidity. The higher the acidity, the lower the pectinesterase activity and the higher both the polymethylgalacturonate lyase and polygalacturonase activities., Centro de Investigación y Desarrollo en Fermentaciones Industriales

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