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1.
Braz. arch. biol. technol ; 63: e20180444, 2020. graf
Article in English | LILACS | ID: biblio-1132252

ABSTRACT

Abstract Fungi have always attracted a lot of attention as they are able to produce a vast repertoire of enzymes that find a broad spectrum of uses in biotechnological and industrial fields. Undoubtedly, one of the most promising biocatalysts is the lipase, which has been widely used for the biotransformation of a number of commercial products due to its high stability, high catalytic efficiency, versatility and selectivity, making it one of the most attractive and best-studied enzymes. In this study we report the isolation and molecular identification of new lipase-producing fungi from different environmental samples from Morocco. The production and activity of extracellular lipases, at different parameters, was evaluated using the Rhodamine B agar, submerged fermentation and biochemical methods. Two fungal strains Arthrographis curvata and Rhodosporidium babjevae, were isolated and found to produce large amounts of lipases. The optimal activity of the extracellular lipase was detected at 40°C and pH 9.0 for A. curvata and at 40 °C and pH 8.0 for R. babjevae. This study add new information at the growing list of fungal species producing lipases with improved physicochemical proprieties which could constitute a new line of research for further studies and to be exploited for industrial or bioremediation purposes.


Subject(s)
Biotechnology , Fungi/enzymology , Lipase/biosynthesis
2.
Braz. j. microbiol ; 49(4): 723-730, Oct.-Dec. 2018. graf
Article in English | LILACS | ID: biblio-974310

ABSTRACT

ABSTRACT The soil represents the main source of novel biocatalysts and biomolecules of industrial relevance. We searched for hydrolases in silico in four shotgun metagenomes (4,079,223 sequences) obtained in a 13-year field trial carried out in southern Brazil, under the no-tillage (NT), or conventional tillage (CT) managements, with crop succession (CS, soybean/wheat), or crop rotation (CR, soybean/maize/wheat/lupine/oat). We identified 42,631 hydrolases belonging to five classes by comparing with the KEGG database, and 44,928 sequences by comparing with the NCBI-NR database. The abundance followed the order: lipases > laccases > cellulases > proteases > amylases > pectinases. Statistically significant differences were attributed to the tillage system, with the NT showing about five times more hydrolases than the CT system. The outstanding differences can be attributed to the management of crop residues, left on the soil surface in the NT, and mechanically broken and incorporated into the soil in the CT. Differences between the CS and the CR were slighter, 10% higher for the CS, but not statistically different. Most of the sequences belonged to fungi (Verticillium, and Colletotrichum for lipases and laccases, and Aspergillus for proteases), and to the archaea Sulfolobus acidocaldarius for amylases. Our results indicate that agricultural soils under conservative managements may represent a hotspot for bioprospection of hydrolases.


Subject(s)
Soil/chemistry , Fungal Proteins/genetics , Archaea/enzymology , Archaeal Proteins/genetics , Fungi/enzymology , Hydrolases/genetics , Soil Microbiology , Soybeans/growth & development , Triticum/growth & development , Brazil , Archaea/isolation & purification , Archaea/classification , Archaea/genetics , Zea mays/growth & development , Agriculture , Metagenome , Metagenomics , Fungi/isolation & purification , Fungi/classification , Fungi/genetics
3.
Braz. j. microbiol ; 49(4): 879-884, Oct.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-1039268

ABSTRACT

ABSTRACT The multi-enzyme complex (crude extract) of white rot fungi Pleurotus ostreatus, Pleurotus eryngii, Trametes versicolor, Pycnosporus sanguineus and Phanerochaete chrysosporium were characterized, evaluated in the hydrolysis of pretreated pulps of sorghum straw and compared efficiency with commercial enzyme. Most fungi complexes had better hydrolysis rates compared with purified commercial enzyme.


Subject(s)
Fungal Proteins/chemistry , Sorghum/chemistry , Cellulases/chemistry , Fungi/enzymology , Lignin/chemistry , Fungal Proteins/metabolism , Plant Stems/microbiology , Plant Stems/chemistry , Sorghum/microbiology , Cellulases/metabolism , Biocatalysis , Fungi/chemistry , Hydrolysis , Lignin/metabolism
4.
Biol. Res ; 51: 28, 2018. tab, graf
Article in English | LILACS | ID: biblio-983933

ABSTRACT

BACKGROUND: Pectinase enzymes catalyze the breakdown of pectin, a key component of the plant cell wall. At industrial level, pectinases are used in diverse applications, especially in food-processing industry. Currently, most of the industrial pectinases have optimal activity at mesophilic temperatures. On the contrary, very little is known about the pectinolytic activities from organisms from cold climates such as Antarctica. In this work, 27 filamentous fungi isolated from marine sponges collected in King George Island, Antarctica, were screened as new source of cold-active pectinases. RESULTS: In semi-quantitative plate assays, 8 out 27 of these isolates showed pectinolytic activities at 15 °C and one of them, Geomyces sp. strain F09-T3-2, showed the highest production of pectinases in liquid medium containing pectin as sole carbon source. More interesting, Geomyces sp. F09-T3-2 showed optimal pectinolytic activity at 30 °C, 10 °C under the temperature of currently available commercial mesophilic pectinases. CONCLUSION: Filamentous fungi associated with Antarctic marine sponges are a promising source of pectinolytic activity. In particular, pectinases from Geomyces sp. F09-T3-2 may be potentially suitable for biotechnological applications needing cold-active pectinases. To the best of our knowledge, this is the first report describing the production of pectinolytic activity from filamentous fungi from any environment in Antarctica.


Subject(s)
Animals , Polygalacturonase/biosynthesis , Porifera/microbiology , Fungi/enzymology , Cold Temperature , Antarctic Regions
5.
Electron. j. biotechnol ; 28: 101-112, July. 2017. ilus, graf, tab
Article in English | LILACS | ID: biblio-1015977

ABSTRACT

Background: The hydrolysis of keratin wastes by microorganisms is considered a biotechnological alternative for recycling and valorization through keratinolytic microorganisms. Despite their resistant structure, keratin wastes can be efficiently degraded by various microorganisms through the secretion of keratinases, which are promising enzymes for several applications, including detergents, fertilizers, and leather and textile industry. In an attempt to isolate keratinolytic microorganisms that can reach commercial exploitation as keratinase producers, the current work assesses the dynamics of keratin biodegradation by several keratinolytic fungal strains isolated from soil. The activity of fungal strains to degrade keratin substrates was evaluated by SEM, FTRIR-ATR spectra and TGA analysis. Results: SEM observations offered relevant information on interactions between microorganism and structural elements of hair strands. FTIR spectra of the bands at 1035­1075 cm-1 assigned to sulfoxide bond appeared because of S­S bond breaking, which demonstrated the initiation of keratin biodegradation. According to TGA, in the second zone of thermal denaturation, where keratin degradation occurs, the highest weight loss of 71.10% was obtained for sample incubated with Fusarium sp. 1A. Conclusions: Among the tested strains, Fusarium sp. 1A was the most active organism in the degradation process with the strongest denaturation of polypeptide chains. Because keratinolytic microorganisms and their enzymes keratinases represent a subject of scientific and economic interest because of their capability to hydrolyze keratin, Fusarium sp. 1A was selected for further studies.


Subject(s)
Fungi/enzymology , Fungi/metabolism , Keratins/metabolism , Peptide Hydrolases/metabolism , Thermogravimetry , Trichoderma/metabolism , Trichophyton/metabolism , Biodegradation, Environmental , Microscopy, Electron, Scanning , Cladosporium/metabolism , Spectroscopy, Fourier Transform Infrared , Fusarium/metabolism , Hydrolysis , Keratins/chemistry , Microsporum/metabolism
6.
An. acad. bras. ciênc ; 89(3,supl): 2327-2340, 2017. tab, graf
Article in English | LILACS | ID: biblio-886786

ABSTRACT

ABSTRACT To the fungal microbiota the UFPE and biotechnological potential enzymatic and antimicrobial production. Air conditioned environments were sampled using a passive sedimentation technique, the air I ratio and the presence of aflatoxigenic strains evaluated for ANVISA. Icelles were to determine the enzymatic activity of lipase, amylase and protease metabolic liquids to determine antimicrobial activity. Diversity was observed in all CAV environments, CFU/m3 ranged from 14 to 290 and I/E ratio from 0.1 to 1.5. The of the fungal genera were: Aspergillus (50%), Penicillium (21%), Talaromyces (14%), Curvularia and Paecilomyces (7% each). Aspergillus sydowii (Bainier & Sartory) Thom & Church presented enzymatic activity and the Talaromyces purpureogenus Samson, Yilmaz, Houbraken, Spierenb., Seifert, Peterson, Varga & Frisvad presented antibacterial activity against all bacteria that all environments present fungal species biodiversity no toxigenic or pathogenic fungi were found, according to ANVISA legislation for conditioned environments and airborne filamentous fungi present potential for enzymatic and antimicrobial activity.


Subject(s)
Aflatoxins/isolation & purification , Biodiversity , Air Microbiology , Fungi/isolation & purification , Fungi/enzymology , Anti-Bacterial Agents/isolation & purification , Universities , Brazil , Public Sector , Aflatoxins/pharmacology , Air Conditioning , Fungi/classification , Anti-Bacterial Agents/pharmacology
7.
Rev. argent. microbiol ; 48(4): 274-278, dic. 2016. ilus, graf, tab
Article in English | LILACS | ID: biblio-1041762

ABSTRACT

Knowledge regarding the enzymatic machinery of fungi is decisive to understand their ecological role. The species of the genus Geastrum are known to grow extremely slowly in pure culture, which makes it difficult to evaluate physiological parameters such as enzyme activity. Qualitative assays were performed on isolates of four species of this genus, showing evidence of laccase, cellulase, pectinase, amylase and lipase activity and suggesting that a wide range of carbon sources can be exploited by these species. For the first time in this genus, quantitative assays verified manganese peroxidase activity (up to 0.6 mU/g) in 30-day old cultures, as well as laccase, β-glycosidase and β-xylosidase activities.


El conocimiento de la maquinaria enzimática de un hongo es decisivo para entender su rol ecológico. Las especies del género Geastrum son conocidas por su crecimiento extremadamente lento en cultivos puros, lo que hace difícil la evaluación de parámetros fisiológicos como las actividades enzimáticas. Se realizaron ensayos cualitativos sobre aislamientos de 4 especies de este género, mostrando evidencias de actividades lacasa, celulasa, pectinasa, amilasa y lipasa, mostrando el amplio rango de fuentes de carbono que pueden ser explotadas por estas especies. Ensayos cuantitativos verificaron por primera vez en este género la actividad manganeso peroxidasa (hasta 0,6 mU/g) en cultivos de 30 días, así como también β-glucosidasa y β-xilosidasa.


Subject(s)
Fungi/enzymology , Xylosidases/isolation & purification , Biotransformation/physiology , Cellulase/isolation & purification , Laccase/isolation & purification , Fungi/physiology , Lipase/isolation & purification
8.
Bol. micol. (Valparaiso En linea) ; 31(1): 19-27, jun. 2016. ilus, tab, graf
Article in Spanish | LILACS | ID: biblio-868808

ABSTRACT

Las amilasas (alfa-amilasa, EC 3.2.1.1 y glucoamilasa, EC 3.2.1.3) son enzimas extracelulares que hidrolizan el almidón en dextrinas hasta glucosa y tienen gran aplicación industrial, especialmente alimentaria; detergentes y en la producción de alcohol a partir de granos. El objetivo del trabajo es seleccionar un hongo filamentoso que presente alta producción de amilasas con características particulares para ser empleadas en biodetergentes. Se estudiaron los siguientes hongos: Penicillium expansum; P. digitatum; P. islandicum; Aspergillus clavatus; A. niger; A. ochraceus; A. fumigatus; A. flavus; A. oryzae; A. nidulans y Geotrichum candidum; Los ensayos se realizaron en un medio de hidrolizado de papa de descarte (variedad Spunta) suplementado con las siguientes sales: KH2 PO4 1,0; NaNO3 3,0; MgSO4 .7H2 O 0,5, a pH 4,0; se inoculó con 2 x106 conidios/mL y se incubaron a 25ºC en un agitador rotatorio a una velocidad de agitación de 250 rpm. Con los extractos enzimáticos parcialmente purificados con (NH4 )2 SO4 al 60 por ciento de saturación, se estudió el efecto del pH (2,5; 3,5; 4,0; 4,5; 5,0; 5,5; 6,0, 7,0 y 8,0) y la temperatura (20; 25; 30; 35 y 40ºC). Los resultados mostraron que la máxima producción de enzima (128 U/L) se obtuvo con Aspergillus niger, en las condiciones ensayadas, a las 48 h de incubación, con alto rendimiento de producto respecto a la biomasa (Yp/x= 18,3 U/g) y productividad volumétrica (Pdv=2,7 U/L). El análisis cualitativo de las enzimas del complejo amilolítico de A. niger mostró que las amilasas implicadas son alfa-amilasa y glucoamilasa y se caracterizaron por hidrolizar en un tiempo de 3 min. manchas mixtas de almidón y grasas de muestras textiles en un rango de pH 4,0 a 8,0 y de 20 a 40 ºC.


The amylases (alpha-amylase, EC 3.2.1.1 and glucoamylase, EC 3.2.1.3) are extracellular enzymes that hydrolyze starch into dextrins to glucose and have great application industrial, especially food, detergents and in the production of alcohol from grains. The objective of the study is to select a filamentous fungus that present high production of amylases showing attractive features to be used in biodetergentes. Were studied following fungus: Penicillium expansum; P. digitatum; P. islandicum; Aspergillus clavatus; A niger; A. ochraceus; A. fumigatus; A. flavus; A. oryzae; A. nidulans and Geotrichum candidum. The tests were conducted in the medium of hydrolyzed potato discard (variety Spunta) supplemented with the following sales: KH2 PO4 , 1.0; NaNO3 , 3.0 and MgSO4 .7H2 O, 0.5, to pH 4.0. Were inoculated with 2 x 106 conidia/ mL and incubated at 25 °C on a rotary Shaker at a speed of 250 rpm. With partially purified enzyme extracts with (NH4 )2 SO4 at 60 percent of saturation, we studied the effect of pH (2.5; 3.5; 4.0; 4.5; 5.0; 5.5; 6.0, 7.0 and 8.0) and temperature (20; 25; 30; 35, and 40 ° C). The results showed that the maximum production of enzyme (128 U/L) was obtained with Aspergillus niger, under the conditions tested, at 48 h of incubation, with high product formation rate with respect to biomass (Yp/x = 18.3 U/g) and volumetric productivity (Pdv = 2,7 U/ hL). The qualitative analysis of the enzymes of the complex amylolític of A. niger showed that involved amylases are α-amylase and glucoamylase and characterized by hydrolyze in 3 min spots mixed starch and fats of textile samples over a range of pH 4.0 to 8.0 and 20 to 40 ° C.


Subject(s)
Amylases/analysis , Aspergillus niger/isolation & purification , Aspergillus niger/enzymology , Fungi/enzymology , Biodegradation, Environmental , Detergents , Hydrogen-Ion Concentration , Hydrolysis , Starch , Temperature
9.
Braz. j. microbiol ; 46(2): 337-346, Apr-Jun/2015. tab
Article in English | LILACS | ID: lil-749736

ABSTRACT

Proteases hydrolyze the peptide bonds of proteins into peptides and amino acids, being found in all living organisms, and are essential for cell growth and differentiation. Proteolytic enzymes have potential application in a wide number of industrial processes such as food, laundry detergent and pharmaceutical. Proteases from microbial sources have dominated applications in industrial sectors. Fungal proteases are used for hydrolyzing protein and other components of soy beans and wheat in soy sauce production. Proteases can be produced in large quantities in a short time by established methods of fermentation. The parameters such as variation in C/N ratio, presence of some sugars, besides several other physical factors are important in the development of fermentation process. Proteases of fungal origin can be produced cost effectively, have an advantage faster production, the ease with which the enzymes can be modified and mycelium can be easily removed by filtration. The production of proteases has been carried out using submerged fermentation, but conditions in solid state fermentation lead to several potential advantages for the production of fungal enzymes. This review focuses on the production of fungal proteases, their distribution, structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications.


Subject(s)
Biotechnology/methods , Fungi/enzymology , Peptide Hydrolases/isolation & purification , Peptide Hydrolases/metabolism
10.
Indian J Exp Biol ; 2014 Nov; 52(11): 1025-1035
Article in English | IMSEAR | ID: sea-153782

ABSTRACT

After cellulose, chitin is the second most abundant organic and renewable polysaccharide in nature. This polymer is degraded by enzymes called chitinases which are a part of the glycoside hydrolase family. Chitinases have many important biophysiological functions and immense potential applications especially in control of phytopathogens, production of chito-oligosaccharides with numerous uses and in treatment and degradation of chitinous biowaste. At present many microbial sources are being explored and tapped for chitinase production which includes potential fungal cultures. With advancement in molecular biology and gene cloning techniques, research on fungal chitinases have made fast progress. The present review focuses on recent advances in fungal chitinases, containing a short introduction to types of chitinases, their fermentative production, purification and characterization and molecular cloning and expression.


Subject(s)
Chitin/metabolism , Chitinases/classification , Chitinases/genetics , Chitinases/isolation & purification , Chitinases/metabolism , Cloning, Molecular , Fermentation , Fungal Proteins/genetics , Fungal Proteins/isolation & purification , Fungal Proteins/metabolism , Fungi/enzymology , Fungi/growth & development , Industrial Microbiology/methods , Mycology/methods
11.
Braz. j. microbiol ; 45(1): 279-286, 2014. graf, tab
Article in English | LILACS | ID: lil-709463

ABSTRACT

Cellulase production was evaluated in two reference strains (T. reesei Rut-C30 and T. reesei QM9414), two strains isolated from a sugarcane cultivation area (Trichoderma sp. IPT778 and T. harzianum rifai IPT821) and one strain isolated in a program for biodiversity preservation in São Paulo state (Myceliophthora thermophila M77). Solid state cultures were performed using sugarcane bagasse (C), wheat bran (W) and/or soybean bran (S). The highest FPA was 10.6 U/gdm for M77 in SC (10:90) at 80% moisture, which was 4.4 times higher than production in pure W. C was a strong inducer of cellulase production, given that the production level of 6.1 U/gdm in WC (40:60) was 2.5 times higher than in pure W for strain M77; T. reesei Rut-C30 did not respond as strongly with about 1.6-fold surplus production. S advantageously replaced W, as the surplus production on SC (20:80) was 2.3 times relative to WC (20:80) for M77.


Subject(s)
Biotechnology/methods , Cellulase/metabolism , Culture Media/chemistry , Fungi/enzymology , Fungi/growth & development , Dietary Fiber/metabolism , Saccharum/metabolism , Sordariales/enzymology , Sordariales/growth & development , Soybeans/metabolism , Trichoderma/enzymology , Trichoderma/growth & development
12.
Article in English | IMSEAR | ID: sea-162952

ABSTRACT

Aim: The study evaluated potential performance of different fungal isolates from agricultural by-products for mannanase production. Study Design: The first experiment, fungal isolates were screened for mannanase production on agar medium containing Locust Bean Gum (LBG) and total fungal count was conducted. In the second experiment, the fungal isolates were further screened for mannanase production in submerged state fermentation. Place and Duration of Study: Microbiology Research Laboratory Federal University of Technology, Akure and Postgraduate Research Laboratory, Obafemi Awolowo University Ile-Ife, Nigeria between September 2011 and March 2012. Methodology: The fungal isolates associated with some agricultural wastes were isolated on LBG containing agar medium by plate assay techniques and counted by standard microbiological methods. Mannanase production was conducted in submerged state fermentation (shaken & static) into which copra meal had been supplemented as the sole carbon source and enzyme activity was determined by dinitrosalicylic acid method. Results: In this study, 11 fungal isolates showed positive results with clear zone around their cultures. Fungal isolate 5A showed the highest activity ratio of 1.8, while the least was observed in isolate 9A12 with activity ratio of 0.64. The highest fungal counts were recorded in fermented coconut with 7.4×102 sfu/g, while cocoa pod and groundnut shell had no fungal growth. In terms of percentage occurrence of fungal isolates from selected agrowastes, it was revealed that Rhizopus japonicus had the highest occurrence of 66.67%, while the same value of 8.33% was observed for Aspergillus fumigatus, A. glaucus, R. stolonifer and Trichosporonoides oedocephalis. In fermentation broth, all the 11 isolates displayed mannanase activity ranging from 0.370 to 21.667 U/ml for static and 0.278 to 3.982 U/ml for shaken condition, with the highest mannanase activity observed with isolate 5A for both culture conditions. According to the cultural characters and microscopic morphology, the isolate 5A being the highest mannanase producer was identified as the Aspergillus fumigatus. Conclusion: In this study, fungal isolates screened and evaluated for mannanase production from agricultural by-products elaborated considerable mannanase activity and this could be exploited for prebiotic preparation.


Subject(s)
Agriculture , Fungi/analysis , Fungi/enzymology , Fungi/isolation & purification , Fungi/metabolism , Fungi/physiology , Industrial Microbiology , Industrial Waste , beta-Mannosidase/biosynthesis
13.
Braz. j. microbiol ; 44(3): 923-926, July-Sept. 2013. ilus, tab
Article in English | LILACS | ID: lil-699782

ABSTRACT

A sensitive and efficient colorimetric method was optimized for detection of esterase enzymes produced by endophytic fungi for development of High-Throughput Screening (HTS). The fungi were isolated and obtained previously from plant species of Cerrado and Atlantic Forest located in areas of environmental preservation in the State of Sao Paulo / Brazil, as part of the project "Chemical and biological prospecting endophytic fungi associated to plant species of Cerrado and Atlantic Forest". The compounds ethyl butyrate, ethyl acetate and methyl propionate were used as standards esters which were hydrolyzed by extracellular enzyme from endophytic fungi (EC. 3.1.1.1 -carboxylesterases) for production of carboxylic acids. Thus, the reduction of the pH increases the protonated indicator concentration (bromothymol blue), changing the color of the reaction medium (from blue to yellow), that can be observed and measured by spectrophotometry at 616 nm. The methodology with acid-base indicator was performed on 13 microorganisms, aiming Periconia atropurpurea asapotential source of esterase for biotransformation of short chain esters. The results also evidenced that this methodology showed to be efficient, fast, cheap, having low consumption of reagents and easy development, and can be applied to screen carboxylic-ester hydrolases in a large number of microorganisms.


Subject(s)
Colorimetry/methods , Endophytes/enzymology , Esterases/analysis , Fungi/enzymology , Acetates/metabolism , Brazil , Butyrates/metabolism , Fungi/isolation & purification , Hydrogen-Ion Concentration , Indicators and Reagents , Plants/microbiology , Propionates/metabolism
14.
Braz. j. microbiol ; 43(4): 1536-1544, Oct.-Dec. 2012. graf, tab
Article in English | LILACS | ID: lil-665841

ABSTRACT

This work is aimed to produce endoglucanase through solid state fermentation in a packed bed bioreactor with the use of the fungus Myceliophtora sp. I-1D3busing a mixture of wheat bran (WB) and sugar cane bagasse (SCB) as culture medium. Preliminary tests were performed in polypropylene plastic bags, controlling the variables temperature (40, 45, and 50ºC), initial moisture content (75, 80, and 85%, w.b.), and weight proportion SCB/WB (1:1, 7:3, and 9:1). The highest enzyme activities in plastic bags were obtained using the substrate proportion of 7:3, 50ºC temperature, and 80% initial moisture content (878 U/grams of dry solid). High activities of filter-paper cellulase and xylanase were also obtained in plastic bags and some results are reported. For the packed bed experiments, the temperature (45 and 50ºC) and the air flow rate (80, 100 and 120L/h) were the controlled variables. Activity of endoglucanase was similar to plastic bag tests. A longitudinal gradient of moisture content, was observed increasing from the bottom to the top of the reactor, even though the longitudinal enzyme activity profile was flat for almost the whole bed. Air flow rate did not affect enzyme activity, while experiments carried out at 50ºC showed higher enzyme activities. The maximum temperature peak observed was at about 6ºC above the process temperature.


Subject(s)
Cellulases/analysis , Fermentation , Fungi/enzymology , Fungi/isolation & purification , Polypropylenes/analysis , Polypropylenes/isolation & purification , Saccharum , Triticum , Xylans/analysis , Food Samples , Industrial Microbiology , Methods , Plastics Industry
15.
Braz. j. microbiol ; 40(4): 790-794, Oct.-Dec. 2009. graf
Article in English | LILACS | ID: lil-528161

ABSTRACT

Enzyme application in biotechnological and environmental processes has had increasing interest due to its efficiency, selectivity and mainly for being environmentally healthful, but these applications require a great volume of enzymes. In this work the effect of different concentrations of ethanol and 2,5 - xylidine on growth and production of laccase by Pycnoporus sanguineus was investigated. In a medium containing 200 mg.L-1 of 2,5 - xylidine or 50 g.L-1 of ethanol, the maximum activity of laccase was 2019 U.L-1 and 1035 U.L-1, respectively. No direct correlation between biomass and activity of laccase was observed for any of the inducers used during the tests. Ethanol concentrations, larger than or equal to 20 g.L-1, inhibited the radial growth of P. sanguineus. This study showed that ethanol, which has less toxicity and cost than the majority of the studied inducers, presents promising perspectives for laccase production by P. sanguineus.


Subject(s)
Biomass , DNA Repair Enzymes/analysis , Ethanol/analysis , Fungi/enzymology , Fungi/genetics , In Vitro Techniques , Laccase/analysis , Enzyme Activation , Methods , Biological Phenomena , Methods
16.
Electron. j. biotechnol ; 12(4): 5-6, Oct. 2009. ilus, tab
Article in English | LILACS | ID: lil-558548

ABSTRACT

Cellulolytic properties of two white rot fungi, Bjerkandera adusta and Pycnoporus sanguineus, cultivated on wheat straw agar medium, were characterized and compared. Optimal growing parameters for maximum enzyme production for both fungi were wheat straw medium pH 5 and 28ºC. B. adusta showed, on the 6th day of culture, carboxymethylcellulose (CMC)ase activity levels 1.6 times higher than maximal P. sanguineus activity, achieved on the 8th day. B. adusta supernatants also displayed higher activity levels towards xylan (3.6-fold) compared to those of P. sanguineus. However, enzymes from P. sanguineus were more robust resisting one hour incubation at high temperatures (up to 80ºC), and exhibiting activity and stability in pH range from 2 to 8. Cellulolytic activities, with molecular masses ranging from 25 to 90 kDa, from the two species were detected in zymograms.


Subject(s)
Enzyme Activation , Cellulose , Fungi/enzymology , Fungi/metabolism , Triticum , Triticum/enzymology , Triticum/metabolism , Electrophoresis, Agar Gel/methods , Culture Media/metabolism , Temperature
17.
Indian J Biochem Biophys ; 2009 Aug; 46(4): 294-298
Article in English | IMSEAR | ID: sea-135208

ABSTRACT

ALP2 gene encoding alkaline protease cloned from Aureobasidium pullulans HN2-3 was ligated into the surface display plasmid and expressed in the cells of the yeast Yarrowia lipolytica. The expressed alkaline protease was immobilized on the yeast cells. The activity of the immobilized enzyme with 6 His tag was found to be significantly higher than that of without 6 His tag. The immobilized enzyme showed lower optimal temperature and a lower affinity for azocasein than the free enzyme purified from A. pullulans HN2-3. The thermal stability of the immobilized enzyme enhanced and the pH stability decreased, compared to that of the free enzyme.


Subject(s)
Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Caseins/chemistry , Cations , Cell Membrane/metabolism , Cloning, Molecular , Endopeptidases/chemistry , Endopeptidases/genetics , Enzymes, Immobilized/chemistry , Fungi/enzymology , Gene Expression Regulation, Fungal , Genetic Engineering/methods , Hydrogen-Ion Concentration , Ions , Kinetics , Models, Biological , Temperature , Yarrowia/enzymology , Yarrowia/genetics
18.
Braz. j. microbiol ; 40(2): 367-372, Apr.-June 2009. ilus, tab
Article in English | LILACS | ID: lil-520225

ABSTRACT

A comparative study on the potential of some biological agents to perform the hydrolysis of stevioside was carried out, aiming at establishing an alternative methodology to achieve the aglycon steviol or its rearranged derivative isosteviol, in high yields to be used in the preparation of novel bioactive compounds. Hydrolysis reactions were performed by using filamentous fungi (Aspergillus niger, Rhizopus stolonifer and Rhizopus arrhizus), a yeast (Saccharomyces cerevisiae) and enzymes (pancreatin and lipases PL250 and VFL 8000). Pancreatin showed the best hydrolytic activity, furnishing isosteviol at 93.9% of yield, at pH 4.0, using toluene as a co-solvent. Steviol was produced using both pancreatin at pH 7.0 (20.2% yield) and A. niger atpH 7 (20.8% yield).


Um estudo comparativo do potencial de alguns agentes biológicos capazes de hidrolisar o esteviosídeo foi realizado,objetivando-se estabelecer uma metodologia alternativa para a obtenção da aglicona esteviol ou seu produto de rearranjo, isoesteviol, em rendimentos elevados que permitam o uso destas agliconas para o preparo de novos compostos bioativos. As reações de hidrólise foram realizadas usando fungosfilamentosos (Aspergillus niger, Rhizopus stolonifer e Rhizopus arrhizus), uma levedura (Saccharomyces cerevisiae) e enzimas(pancreatina, lipase PL250 e lipase VFL 8000). A pancreatina mostrou a melhor atividade hidrolítica dentre os sistemastestados, fornecendo isoesteviol com rendimento de 93,9% em pH 4,0, usando tolueno como co-solvente. Esteviol foi produzido tanto usando pancreatina em pH 7,0 (20,2% derendimento) quanto usando A. niger em pH 7,0 (20,8% de rendimento).


Subject(s)
Biological Reactions , Fungi/enzymology , Fungi/isolation & purification , Lipase/analysis , Pancreatin/analysis , Stevia/enzymology , Chromatography, High Pressure Liquid , Hydrolysis , Methods , Methods
19.
Electron. j. biotechnol ; 11(4): 13-14, Oct. 2008. ilus, tab
Article in English | LILACS | ID: lil-531921

ABSTRACT

Four white rot fungi (WRF) strains, Phanerochaete chrysosporium, Trametes versicolor, Coriolopsis polyzona and Pycnoporus coccineus, were tested for efficiency of treatment of Olive Oil mill wastewaters (OOMW) in relation with their cultivation mode, i.e. under the form of free mycelium, mycelium immobilized in alginate beads and solid state cultivation on Petri dishes. Study of biodegradation of phenolic compounds, chemical oxygen demand (COD) decrease and decolourisation of OOMW have shown that Coriolopsis polyzona and Pycnoporus coccineus degradation performances were apparently only slightly affected by the cell cultivation procedures experienced here. In contrast, Phanerochaete chrysosporium and Trametes versicolor showed respectively marked preferences for solid state and alginate immobilisation procedures. Both mono and polyphenolics were reduced to different extent during incubation depending on the strain, as shown by gel filtration analysis. Final pH obtained after fungal treatment of the OOMW based medium (initial pH of 5.0) was measured in order to evaluate the possibility of releasing friendly the treated wastewater in the environment. Laboratory studies as reported here may be useful for orienting the choice of a strain for treating pollution by OOMW in a particular real situation.


Subject(s)
Basidiomycota/enzymology , Fungi/enzymology , Phanerochaete/enzymology , Water Purification/methods , Alginates , Biodegradation, Environmental/methods , Peroxidases , Vegetable Fats
20.
Braz. j. microbiol ; 39(1): 122-127, Jan.-Mar. 2008. graf, tab
Article in English | LILACS | ID: lil-480687

ABSTRACT

Cellulase is a complex enzyme system, commercially produced by filamentous fungi under solid-state and submerged cultivation. It has wide applicability in textile, food and beverage industry for effective saccharification process. In this study, cellulolytic enzyme activity, particularly endoglucanase of 26 Streptomyces strains isolated from garden soil was examined, including two isolates selected on the basis of potential cellulolytic activity on Bennett's agar medium. To enhance the endoglucanase formation in broth culture, different conditions including carbon and nitrogen sources, and growth conditions were tested. The maximum endoglucanase activity (11.25-11.90 U/mL) was achieved within 72-88 h in fermentation medium containing Tween-80, followed by phosphate sources. Both cellulolytic Streptomyces isolates gave almost equal quantity of enzyme in all trials. However the effect of medium ingredients on endoglucanase induction diverged with strains in some extent.


A celulase é um sistema enzimático complexo, produzido comercialmente a partir de fungos filamentosos através de cultivo em estádio sólido e submerso. Tem uma grande aplicação na indústria têxtil e de alimentos e bebidas no processo de sacarificação. Nesse estudo, examinou-se a atividade celulolítica, especialmente de englucanase, de 26 cepas de Streptomyces isoladas de solo, incluindo duas cepas selecionadas por sua atividade celulolítica no ágar Bennett. Para estimular a produção de englucanase em meio de cultura, diferentes condições de cultivo, incluindo fonte de carbono e nitrogênio e condições de crescimento, foram avaliadas. A atividade máxima de glucanase (11,25 a 11,90 U/mL) foi obtida em 72-88h em meio de cultura contendo Tween-80, seguido por fontes de fosfato. Ambas as cepas celulolíticas de Streptomyces produziram quase a mesma quantidade de enzima em todos os experimentos. Entretanto, o efeito dos ingredientes do meio na indução da glucanase divergiu de acordo com a cepa.


Subject(s)
Clinical Enzyme Tests , Cellulases/analysis , Fungi/enzymology , Fungi/isolation & purification , In Vitro Techniques , Culture Media/isolation & purification , Streptomyces/enzymology , Streptomyces/isolation & purification , Fermentation , Methods , Soil , Textile Industry
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