Biomass of unconventional plants from Brazilian semiarid as substrate for hydrolytic enzymes production by Aspergillus niger under solid and submerged fermentation

Aspergillus niger KIJH was grown in solid and submerged fermentation using leaves and roots (with and without bark) of plants typically from Brazilian semiarid as substrate to produce a multienzymatic extract, which was characterised for its potential biotechnological applications. Solid-state fermentation (SSF) was applied to select the most promising plants biomass as induction substrates for the production of hydrolytic enzymes by fungus. The best biomasses were used as substrate in submerged fermentation (SmF) assays at two scales. Samples of up scale fermented culture were partially purified by ultrafiltration and activity and pH and temperature stability of CMCase and xylanase were evaluated. A. niger KIJH produced hydrolytic enzymes under SSF containing unconventional plants biomass from Brazilian semiarid. In SmF conditions, maximum CMCase (0.264 U mL-1) and xylanase (1.163 U mL-1) activities were induced by Jacaratia corumbensis. Scaling up the SmF to 500 mL of medium was able to maintain constant the production of CMCase (0.346 U mL-1) and xylanase (1.273 U mL-1) on the fermented culture. Ultrafiltered and concentrated extract presented CMCase activities practically constant in all temperature ranges (30-80°C) and pH (3.0-9.0), while xylanase optimum activity temperature was 50°C and pH in the range of 3.0 to 5.0. CMCase activity remained stable for 24 hours at 50°C

Efecto antifúngico y antimicotoxigénico de Lactobacillus plantarum CRL 778 a diferentes valores de actividad de agua / Antifungal and antimycotoxigenic effect of Lactobacillus plantarum CRL 778 at different water activity values

Ochratoxin A (OTA) is a mycotoxin produced by filamentous fungi with high impact Lactic acid bacteria; in food safety due to its toxicity. In the last decade, the presence of OTA was widely reported in different foods. In this study, the ability of Lactobacillus (L.) plantarum CRL 778 to control growth and OTA production by Aspergillus (A.) niger 13D strain, at different water activity (a w) values (0.955, 0.964, 0.971, 0.982, and 0.995) was determined in vitro. Both parame ters were significantly (p<0.05) reduced by the lactobacilli and the effect depended on a w. Greatest growth rate inhibition (46.9%) was obtained at a w = 0.995, which is the most suitable value for growth and production of antifungal metabolites (lactic acid, acetic acid, phenyllac-tic and hydroxyl-phenyllactic acids) by L. plantarum CRL 778. Besides, morphological changes and inhibition of melanin synthesis were observed in colonies of A. niger 13D in presence of L. plantarum CRL 778 at a w ranged between 0.971 and 0.995. In addition, maximum reduction (90%) of OTA production took place at a w = 0.971, while inhibition of fungi growth was more evident at a w =0.995. These findings suggest that L. plantarum CRL 778 could be used for control of ochratoxigenic fungal growth and OTA contamination in different fermented foods with a w values between 0.971 and 0.995.

Ocratoxina A (OTA) es una micotoxina producida por hongos filamentosos con un alto impacto en la seguridad alimentaria debido a su toxicidad. En la última década se ha reportado ampliamente a nivel mundial, la presencia de OTA en diversos alimentos. En este estudio se evaluó in vitro, la capacidad de Lactobacillus (L.) plantarum CRL 778 de controlar el crecimiento y la producción de OTA por Aspergillus (A.) niger 13D, a diferentes valores de actividad de agua (a w): 0.955, 0.964, 0.971,0.982 y 0.995). La cepa láctica redujo significativamente (p <0.05) ambos parámetros, siendo el efecto dependiente del valor de a w. La mayor inhibición del crecimiento (46.9%) se obtuvo a a w =0.995, valor más adecuado para el crecimiento y producción de metabolitos antifúngicos (ácido láctico, ácido acético, ácidos fenil-láctico e hidroxi-fenil láctico) por la cepa láctica. Además, se observaron cambios morfológicos en las colonias de A. niger 13D, crecidas en presencia de L. plantarum CRL 778 a valores de a w de 0.971 y 0.995. El porcentaje máximo de reducción en la producción de OTA (90%) por la cepa láctica se observó a un valor de a w = 0.971, mientras la inhibición del crecimiento fúngico fue mayor cuando a w = 0.995. Estos hallazgos sugieren que L. plantarum CRL 778 podría emplearse para el control de la contaminación por hongos ocratoxigénicos en alimentos con valores de aw comprendidos entre 0.971-0.995.

Optimizing culture conditions for production of intra and extracellular inulinase and invertase from Aspergillus niger ATCC 20611 by response surface methodology (RSM)

Abstract The aim of this study was obtain a model that maximizes growth and production of inulinase and invertase by Aspergillus niger ATCC 20611, employing response surface methodology (RSM). The RSM with a five-variable and three-level central composite design (CCD) was employed to optimize the medium composition. Results showed that the experimental data could be appropriately fitted into a second-order polynomial model with a coefficient of determination (R2) more than 0.90 for all responses. This model adequately explained the data variation and represented the actual relationships between the parameters and responses. The pH and temperature value of the cultivation medium were the most significant variables and the effects of inoculum size and agitation speed were slightly lower. The intra-extracellular inulinase, invertase production and biomass content increased 10-32 fold in the optimized medium condition (pH 6.5, temperature 30 °C, 6% (v/v), inoculum size and 150 rpm agitation speed) by RSM compared with medium optimized through the one-factor-at-a-time method. The process development and intensification for simultaneous production of intra-extracellular inulinase (exo and endo inulinase) and invertase from A. niger could be used for industrial applications.

Remediación de residuos sólidos contaminados con Cr(vi) por un hongo filamentoso / Remediation of waste contaminated with Cr (VI) by a filamentous fungus

La biotecnología ambiental recurre a organismos capaces de reducir los niveles de metales pesados, entre ellos el Cr(VI), contenido en residuos y efluentes agroindustriales. El objetivo del trabajo fue estudiar la biorremediación de un residuo como pulpa de limón contaminada con Cr(VI) y el efecto del metal sobre el crecimiento fúngico. Se utilizaron tres hongos filamentosos como Aspergillus niger; Penicillium expansum y P. islandicum para remediar pulpa de limón (residuo) contaminada con Cr(VI) (50 mg/L) que se realizó en las siguientes condiciones de cultivo: la pulpa se suplementó con urea, 0,006; (NH4) 2SO4, 0,012; KH2PO4,0,003 y KCl, 0,001 g/ g; 105 conidios/g, a pH 2,5, 30°C y 96 h de incubación. Se estudió el efecto tóxico de diferentes concentraciones (5; 10; 20 y 50 mg/L) del metal sobre el desarrollo del hongo de mayor eficiencia de remediación (Ef. por ciento). Aspergillus niger; obtuvo mayor EF. de remediación (97 por ciento) respecto a Penicillium expansum (95 por ciento) y P. islandicum (94 por ciento), del residuo contaminado con 50 mg/L de Cr(VI). Se determinó que la presencia de Cr(VI) y no su concentración estimuló la maduración temprana (48 h) de los conidios (blancos a negros) de A. niger, sin que se observe alteraciones en el micelio con respecto al control (72h), desarrollado en la pulpa sin el metal. En conclusión, A. niger fue más resistente y presentó altas Ef. de remediación de Cr(VI) de residuos sólidos, este proceso es una alternativa a las tecnologías físico-químicas, debido que los microorganismos pueden remover selectivamente diferentes iones de zonas contaminadas.

Environmental biotechnology uses organisms capable of reducing levels of heavy metals, including the Cr (VI), contained in waste and agro-industrial effluents. The objective of this work was to study bioremediation of waste contaminated with Cr(VI) lemon pulp and the effect of the metal on the fungal growth. We used three filamentous fungi such as Aspergillus niger; Penicillium expansum and P. islandicum to remedy pulp from lemon (residue) contaminated with Cr(VI) (50 mg/L) that was conducted in the following conditions of cultivation: the pulp is supplemented with urea, 0.006; (NH4) 2SO4, 0.012; KH2PO4, 0.003 and KCl, 0.001 g / g; 105 conidia/g, at pH 2.5, 30 ° C and 96 h of incubation. We studied the toxic effect of different concentrations (5, 10, 20 and 50 mg/L) of the metal on the development of the fungus increased efficiency of remediation (Ef. percent). Aspergillus niger; obtained greater EF. remediation (97 percent) with respect to Penicillium expansum (95 percent) and P. islandicum (94 percent), 50 mg/L of Cr (VI)-contaminated waste. It was determined that the presence of Cr (VI) and not its concentration stimulated early maturation (48 h) of conidia (white on black) from A. niger, unless you observe alterations in the mycelium as compared to the control (72 h), developed in the pulp without the metal. In conclusion, A. niger was stronger and presented high Ef. remediation of Cr (VI) waste, this process is an alternative to physico-chemical technologies, due to the micro-organisms be removed selectively different ions from contaminated areas.

Screening of thermotolerant and thermophilic fungi aiming β-xylosidase and arabinanase production

Plant cell wall is mainly composed by cellulose, hemicellulose and lignin. The heterogeneous structure and composition of the hemicellulose are key impediments to its depolymerization and subsequent use in fermentation processes. Thus, this study aimed to perform a screening of thermophilic and thermotolerant filamentous fungi collected from different regions of the São Paulo state, and analyze the production of β-xylosidase and arabinanase at different temperatures. These enzymes are important to cell wall degradation and synthesis of end products as xylose and arabinose, respectively, which are significant sugars to fermentation and ethanol production. A total of 12 fungal species were analyzed and 9 of them grew at 45 ºC, suggesting a thermophilic or thermotolerant character. Additionally Aspergillus thermomutatus anamorph of Neosartorya and A. parasiticus grew at 50 ºC. Aspergillus niger and Aspergillus thermomutatus were the filamentous fungi with the most expressive production of β-xylosidase and arabinanase, respectively. In general for most of the tested microorganisms, β-xylosidase and arabinanase activities from mycelial extract (intracellular form) were higher in cultures grown at high temperatures (35-40 ºC), while the correspondent extracellular activities were favorably secreted from cultures at 30 ºC. This study contributes to catalogue isolated fungi of the state of São Paulo, and these findings could be promising sources for thermophilic and thermotolerant microorganisms, which are industrially important due to their enzymes.

Bioremediation of dyes by fungi isolated from contaminated dye effluent sites for bio-usability

Biodegradation and detoxification of dyes, Malachite green, Nigrosin and Basic fuchsin have been carried out using two fungal isolates Aspergillus niger, and Phanerochaete chrysosporium, isolated from dye effluent soil. Three methods were selected for biodegradation, viz. agar overlay and liquid media methods; stationary and shaking conditions at 25 °C. Aspergillus niger recorded maximum decolorization of the dye Basic fuchsin (81.85%) followed by Nigrosin (77.47%), Malachite green (72.77%) and dye mixture (33.08%) under shaking condition. Whereas, P. chrysosporium recorded decolorization to the maximum with the Nigrosin (90.15%) followed by Basic fuchsin (89.8%), Malachite green (83.25%) and mixture (78.4%). The selected fungal strains performed better under shaking conditions compared to stationary method; moreover the inoculation of fungus also brought the pH of the dye solutions to neutral from acidic. Seed germination bioassay study exhibited that when inoculated dye solutions were used, seed showed germination while uninoculated dyes inhibited germination even after four days of observation. Similarly, microbial growth was also inhibited by uninoculated dyes. The excellent performance of A. niger and P. chrysporium in the biodegradation of textile dyes of different chemical structures suggests and reinforces the potential of these fungi for environmental decontamination.

Utilization of molasses and sugar cane bagasse for production of fungal invertase in solid state fermentation using Aspergillus niger GH1

Agro-industrial wastes have been used as substrate-support in solid state fermentation for enzyme production. Molasses and sugarcane bagasse are by-products of sugar industry and can be employed as substrates for invertase production. Invertase is an important enzyme for sweeteners development. In this study, a xerophilic fungus Aspergillus niger GH1 isolated of the Mexican semi-desert, previously reported as an invertase over-producer strain was used. Molasses from Mexico and Cuba were chemically analyzed (total and reducer sugars, nitrogen and phosphorous contents); the last one was selected based on chemical composition. Fermentations were performed using virgin and hydrolyzate bagasse (treatment with concentrated sulfuric acid). Results indicated that, the enzymatic yield (5231 U/L) is higher than those reported by other A. niger strains under solid state fermentation, using hydrolyzate bagasse. The acid hydrolysis promotes availability of fermentable sugars. In addition, maximum invertase activity was detected at 24 h using low substrate concentration, which may reduce production costs. This study presents an alternative method for invertase production using a xerophilic fungus isolated from Mexican semi-desert and inexpensive substrates (molasses and sugarcane bagasse).

Screening the antifungal activity of essential oils against decay fungi from palmyrah leaf handicrafts

BACKGROUND: The whitish tender leaves of Palmyrah are used for making handicrafts. The problem with these articles is discolouration with time and become more brittle due to fungal attack. This could be prevented by some protective coating. Instead of expensive and harmful chemicals we decided to test natural plant essential oils to control fungal attack. Palmyrah leaf article decay fungi were isolated from two different sites of Jaffna peninsula. In this investigation Antifungal Activity of different plant essential oils from neem (Azadirachta indica), castor (Ricinus communis), citronella (Cymbopogon sp) and camphor (Cinnamomum camphora) obtained from local market have been evaluated against isolated fungi. For screening of Antifungal activity, tests and controls were set to determine minimum inhibitory concentration (MIC) and Percentage of Growth Inhibition. RESULTS: Morphologically three different types of Palmyrah leaf decay fungi were isolated and characterized asAspergillus niger, Aspergillus flavus and Penicillium sp. Neem and castor oils have recorded no significant (0.05 > P) antifungal activity while citronella and camphor oils showed significantly different antifungal activity compared with control. Camphor oil and Citronella oil showed 100, 58.13% of average growth inhibition for A. niger. 96.38, 51.32% for A.flavus and 84.99, 72.76% forPenicillium sp respectively. Camphor oil showed highest percentage of growth inhibition at lowest minimum inhibitory concentration compared with citronella oil. Camphor oil was found to be highly antifungal and most effective against A niger, and A. flavus, compared with Penicillium sp and gave 100 percentage of growth inhibitions at 5, 1 and 15 ml/dl minimum inhibitory concentration respectively. CONCLUSION: Significantly higher broad-spectrum of antifungal activity was observed in camphor oil than other tested oils because it showed highest percentage of growth inhibition at lowest inhibitory concentration. Therefore it could be used for the development of new environmental friendly antifungal agent for the preservation of leafy handicrafts. Further formulation, field experiments are necessary to achieve this target.

Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds

Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 x 10(5) cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH)2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained.

Producción de fructooligosacáridos por invertasa de Aspergillus niger IB56: un prebiótico de importancia para la salud humana y animal / Production of fructooligosaccharides by invertase of Aspergillus niger: a prebiotic of importance for the human health and animal

A partir de jarabe de fructosa se aislaron e identificaron microorganismos productores de invertasa. Aspergillus niger IB56 fue el que produjo mayor concentración de la enzima con actividad transferasa (5,6U/ml). Se estudió la producción de fructooligosacáridos (FOS) a diferentes pH (3,0; 4,0; 4,5; 5,0 y 5,5); temperaturas (20, 25, 30 y 40 ºC), concentración de sacarosa (150; 300 y 450 g/l) y tiempos de incubación (60; 90 y 120 min.). La máxima producción de FOS (105 g/l) se obtuvo con una concentración de sacarosa de 300 g/l; a pH 5,0; temperatura 20ºC y a los 60 min de incubación. La enzima invertasa posee especificidad para producir FOS como 1-cestosa y nistosa, prebióticos de importancia en la industria farmacéutica porque tienen efectos benéficos sobre la salud y estimulan la flora microbiana del intestino humano y animal como Lactobacillus y Bifidobacterium.

Several microorganisms that produce invertase were isolated from fructose syrup and identified. Aspergillus nigerIB56 was the one that produced the greatest concentration of the enzyme with transferase activity (5.6 U/ml). We studied the production of fructooligosaccharides (FOS) at different pH (3.0, 4.0, 4.5, 5.0 and 5.5), temperatures (20, 25, 30 and 40ºC), sucrose concentrations (150, 300 and 450 g/l) and incubation times (60, 90 and 120 min.). Maximum FOS production (105 g/l) was obtained with a sucrose concentration of 300 g/l, pH 5.0, at 20 ºC after 60 min of incubation. The enzyme invertase specifically produces FOSsuch as 1-kestose and nistose, which are important prebioticsin the pharmaceutical industry because they have beneficial health effects and stimulate the intestinal microbial flora such as Lactobacillus and Bifidobacterium in humans and animals.

Activity of Tri-N-Butyl Tin maleate in carpets against Staphylococcus aureus and Aspergillus niger, verified through two methodologies: Inhibition Halo (HZ) and Inhibition Surface (Print) / Atividade de tri-n-butyl tin maleate em carpetes contra Staphylococcus aureus e Aspergillus niger, verificada através de duas metodologias: Zona de Inibição (ZI) e Superfície de Inibição (Impressão)

The aim of the present study was to verify the activity of the Tri-N-Butyl Tin maleate compound against Staphylococcus aureus and Aspergillus niger, after its industrial application in 40 samples of carpets of different materials (polypropylene, polyester, polyamide and wool). The qualitative assays were performed through two methodologies: Inhibition Halo (HZ) and Inhibition of Surface (Print). The carpet with the product inhibited 100 percent of bacterial (Staphylococcus aureus) and fungi (Aspergillus niger) growth, under the conditions of this study. The microbial inhibition was higher in upper portion of carpets. The methodologies employed appear to be adequate to test the bactericide and fungicide activities of the Tri-N-Butyl Tin maleate. The print methodology confirmed the results obtained by the inhibition zone assay. Further studies using the same methodologies are needed to confirm our results.

O objetivo do presente estudo foi verificar a atividade do composto maleato de estanho tri-n-butílico contra Staphylococcus aureus e Aspergillus niger, após sua aplicação industrial em 40 amostras de carpetes de diferentes materiais (polipropileno, poliéster, poliamida e lã). Os ensaios qualitativos foram realizados através de duas metodologias: Zona de Inibição (ZI) e Superfície de Inibição (Impressão). Os carpetes tratados com o produto apresentaram 100 por cento de inibição de crescimento bacteriano (Staphylococcus aureus) e fúngico (Aspergillus niger), sob as condições desse estudo. A inibição de crescimento microbiano foi mais elevada na porção superior dos carpetes. As metodologias empregadas parecem ser adequadas para testar a atividade bactericida e fungicida do maleato de estanho tri-n-butílico. A metodologia de impressão confirmou os resultados obtidos no ensaio de zona de inibição. Estudos futuros utilizando as mesmas metodologias são necessários para confirmação destes dados.

Isolation, characterization and antifungal activity of Streptomyces sampsonii GS 1322.

For new antifungal antibiotics from actinomycetes, a strain of Streptomyces GS 1322 was isolated from a sample of garden soil. The strain was found to possess antagonistic activity against four fungi i.e., Candida albicans, Aspergillus niger, Microsporum gypseum and Trichophyton sp. The strain was identified as Streptomyces sampsonii and the antifungal compound produced by it was found to be the heptaene group of polyene antibiotics.

Physiological, morphological, and mannanase production studies on Aspergillus niger uam-gs1 mutants

Mutant strains from Aspergillus niger UAM-GS1 were produced by UV radiation to increase their hemicellulolytic and cellulolytic activity production. The mutant strains showing more enzymatic activity were those labelled GS1-S059 and GS1-S067. These strains also showed the largest relationship between diameter of hydrolysis zone and colony diameter. The mutant GS1-S067 showed a colony radial extension rate and a biomass growth rate g biomass/(cm² h), 1.17 times higher than that achieved by strain UAM-GS1. The high invasive capacity makes this mutant strain a promising alternative for its use in solid substrate fermentation (SSF). The morphological properties of the two mutant strains were evaluated by using scanning electron microscopy. The diameter of the sporangium of the mutant strains GS1-S059 and GS1-S067 was significantly larger (P < 0.05) than that found for the parental strain. The hypha length and diameter of the mutant strains significantly changed (P < 0.05) compared to the parental strain. A Pearson correlation analysis on hypha length, sporangium diameter, and cellulase and xylanase activities indicated that there was a strong relationship among these variables in relation to mannanase activity. Mutant strains GS1-S059 and GS1-S067 significantly increased their level of mannanase, xylanase and cellulase production, compared to the parental strain, improving their potential industrial applications.

Biosorption of Baftkar textile effluent.

Decolourization of wastewater from a textile plant by a marine Aspergillus niger was studied. The fungus was previously isolated from Gorgan Bay in the Caspian Sea. The kinetics of decolourization was studied by varying energy sources. The best decolourization was achieved when sucrose was used as source of carbon and energy. NH4+ ion was demonstrated to be the best nitrogen source. Color reduction was found to increase from 80-97% as inoculum concentration increased from 0.04-1.0 g/L. A minimum inoculum of 0.2 g/L is necessary to achieve decolourization. The optimal temperature for the growth of A. niger on Baftkar wastewater is found to be 30 degrees C. 90-96% colour reduction is achieved in 19-20 hr of contact of mycelium cell with the wastewater. Colour reduction in a continuous column reactor of 70% was obtained using treated mycelium (NaOH, 90 degrees C) after 1 hr.

Economic utilization of agro-industrial wastes through solid state fermentation by aspergillus niger f-21 for alpha-amylase production

Solid state fermentation technique was carried out in utilization of various agro-industrial wastes from different industries for the production of fungal alpha-amylase [E.C. 3.2.1.1] by locally isolated strain of Aspergillus niger. Culture conditions affect enzyme formation such as incubation time, initial pH, moisture level, moistening agent and nitrogen source were investigated. High level of enzyme activity could be obtained after 48 hr using potato peels as a substrate with initial pH 6.0 using whey permeate level of 50% as a moistening agent and urea as a cheap nitrogen source at level 400 mg N/1 moistening agent. The obtained enzyme has an activity of 620 U/g fermented substrate. The obtained enzyme was stable in a wide range of pH between 3-9 as well as the enzyme proceeds through wide range of temperature ranged between 50-85C. The enzyme looses only about 10% from its optimal activity between 90-100C and at 105C the relative activity was about 27%. The crude enzyme was heat stable when incubated at 70C for 39 hr retaining its maximum activity. Crude enzyme was applied for potato starch digestion using different slurries concentrations up to 30% giving about 96% digestion

Metabolism of polycyclic aromatic hydrocarbons by Aspergillus niger.

Aspergillus niger metabolised polycyclic aromatic hydrocarbons such as naphthalene, anthracene and phenanthrene. The maximum weight of mycelium was obtained at 8-10 days of incubation. TLC and HPLC analysis revealed the accumulation of metabolites in resting cell culture. The metabolism of naphthalene and anthracene follows the gentisate pathway, where as phenenthrene was metabolised via protocatechuate in this organism.

Effect of environmental conditions on the productivity and activity of proteases from aspergillus niger and aspergillus terreus grown on whey

Nine-day old culture of A. niger and A. terreus achieved the maximum yield of proteases in response to initial pH 5.25 and 6.15, respectively. Maximum activity was recorded at pH 8 for alkaline proteases from both fungi whereas acid protease from A. niger expressed maximum activity at pH 5. Alkaline proteases from A. niger and A. terreus expressed 2 pKa values at pH 6.1 or 8.9 and pH 7.0 or 8.7, respectively; while, acid protease from A. niger exhibited these values at pH 4.2 and 6.6. Phosphate buffer at 0.02-0.1 M favored the accumulation of the enzyme in the culture of A. niger, a phenomenon that was observed with A. terreus at 0.01 M or in the absence of phosphate buffer. The temperature optima for the formation of proteases were 25 and 28C for A. niger and A. terreus, respectively. However, the biosynthesis of proteases was always associated with the growth of both fungi. Temperature optima were 50, 40 and 40C for alkaline proteases from A. niger and A. terreus and acid protease from A. niger, respectively. Activation energies were 17.0, 6.9 and 12.6 kJ mol-1 for the above-mentioned enzymes in the same sequence indicating that protease from different Aspergilli or even from the same individual have different properties

Optimal conditions for scp production by mixed cultures of A. niger and Cr. laurentti grown in sugar cane vinasse medium

Foi estudado o crescimento da cultura mista de Aspergillus niger e Cryptococcus laurentii em vinhaça, em frascos agitados, para a otimizaçäo das proporçöes ideais de carbono, nitrogênio e fósforo (20 a 30:3.0:0.1); concentraçäo de carboidrato (3 a 32 g/l); pH inicial (3,5 a 5,5); temperatura (25 a 35oC) e tempo de incubaçäo, objetivando produçäo de biomassa e depuraçäo biológica da vinhaça, um resíduo de destilarias. A adiçäo de nitrogênio e fósforo ao meio de vinhaça alterou significativamente a produçäo de biomassa, o consumo de carboidrato e a reduçäo de DBO. No entanto, com concentraçöes maiores que 3 g/l de carboidrato no meio de cultura, o conteúdo proteíco da biomassa aumentou. O pH inicial do meio de cultura e a temperatura de incubaçäo näo alteraram a reduçäo de DBO que permaneceu por volta dos 50 por cento. O pH da vinhaça esteve sempre jperto de 7.0 depois de 48 horas de cultivo. O crescimento do cultivo misto alcançou a máxima produçäo de biomassa depois de 24 horas, com os melhores resultados para reduçäo de DBO (cerca de 80 por cento) e teor de proteína (40 por cento) após 72 horas em meio otimizado (20:3.0:0.1 pa C:N:P, 8g/l de carboidrato, pH 4,6 e 30oC para incubaçäo). Os resultados obtidos, quando comparados com os dados dos cultivos puros, näo sugerem ainda a utilizaçäo do cultivo misto de A. niger + Cr. laurentii para a produçäo de SCP a partir de vinhaça, pois a concentraçäo em biomassa foi baixa

Production of cellulases and beta-glucosidase by new isolate of Aspergillus niger F-92

The production of cellulase was investigated in shakes culture of a new isolate of Aspergillus niger F-92. Amongst different concentration of cellulose powder tested, concentration 1.5% [w/v] showed maximum production of enzyme after nine days incubation at 30C. The optimum initial pH for enzyme production was found to be 5.0. The production of cellulases was sensitive to nitrogen source in the medium, the optimum production was seen when soybean flour [600 mg N/L] was used as the only nitrogen source. KH2PO4 [0.2% w/v] was found to be optimum at concentration 0.3 M caused marked increase in cellulases and beta-glucosidase production. Under the optimum culture conditions the activities of FPase, CMCase and beta-glucosidase were 1.89, 4.2 and 7.82 IU/ml, respectively