Simple method for determining cellulolytic activity in fungi
Wild fungal strains isolated from litter and soil were inoculated in test tubes containing a synthetic medium with 0.5% sodium carboxymethylcellulose (CMC) as sole carbon source. After the incubation time, cylindrical probes were taken from each tube using a borer and stained with Congo red 0.1% to...
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1997
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Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03257541_v29_n4_p210_Magnelli http://hdl.handle.net/20.500.12110/paper_03257541_v29_n4_p210_Magnelli |
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paper:paper_03257541_v29_n4_p210_Magnelli2023-06-08T15:32:43Z Simple method for determining cellulolytic activity in fungi Magnelli, Paula E. Martinez, Alicia Elba Mercuri, Oscar Alberto carboxymethylcellulose cellulase coloring agent congo red fungal protein article comparative study Deuteromycetes enzymology fungus metabolism methodology microbiology mycology sensitivity and specificity species difference Carboxymethylcellulose Cellulase Coloring Agents Congo Red Fungal Proteins Fungi Mitosporic Fungi Mycology Sensitivity and Specificity Soil Microbiology Species Specificity Wild fungal strains isolated from litter and soil were inoculated in test tubes containing a synthetic medium with 0.5% sodium carboxymethylcellulose (CMC) as sole carbon source. After the incubation time, cylindrical probes were taken from each tube using a borer and stained with Congo red 0.1% to reveal the remaining CMC. The relative cellulase activity was estimated by measuring the deepness of the clearing zone, and the growth by the limit of mycelial penetration in the medium. The ratio: hydrolysis zone/depth of growth, provides additional information to compare strains. A test using culture supernatants of liquid media growing strains was developed for enzyme assay using a modification of the cylinder-plate method. Petri dishes were filled with agar-CMC medium and cups were cut off with a steel punch. Such cups were filled with culture supernatants. After incubation, the plated were stained with Congo red, and the diameter of each cleared zone was recorded. Comparative studies of strains could be performed as well as quantitation of enzyme activity using a standard cellulase solution and computing the area of the cleared zones. This method may be useful when a large number of strains must be tested for cellulolytic activity or when conventional tests fail for assaying enzymatic induction in vitro. Fil:Magnelli, P.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Martínez, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Mércuri, O.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1997 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03257541_v29_n4_p210_Magnelli http://hdl.handle.net/20.500.12110/paper_03257541_v29_n4_p210_Magnelli |
institution |
Universidad de Buenos Aires |
institution_str |
I-28 |
repository_str |
R-134 |
collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
topic |
carboxymethylcellulose cellulase coloring agent congo red fungal protein article comparative study Deuteromycetes enzymology fungus metabolism methodology microbiology mycology sensitivity and specificity species difference Carboxymethylcellulose Cellulase Coloring Agents Congo Red Fungal Proteins Fungi Mitosporic Fungi Mycology Sensitivity and Specificity Soil Microbiology Species Specificity |
spellingShingle |
carboxymethylcellulose cellulase coloring agent congo red fungal protein article comparative study Deuteromycetes enzymology fungus metabolism methodology microbiology mycology sensitivity and specificity species difference Carboxymethylcellulose Cellulase Coloring Agents Congo Red Fungal Proteins Fungi Mitosporic Fungi Mycology Sensitivity and Specificity Soil Microbiology Species Specificity Magnelli, Paula E. Martinez, Alicia Elba Mercuri, Oscar Alberto Simple method for determining cellulolytic activity in fungi |
topic_facet |
carboxymethylcellulose cellulase coloring agent congo red fungal protein article comparative study Deuteromycetes enzymology fungus metabolism methodology microbiology mycology sensitivity and specificity species difference Carboxymethylcellulose Cellulase Coloring Agents Congo Red Fungal Proteins Fungi Mitosporic Fungi Mycology Sensitivity and Specificity Soil Microbiology Species Specificity |
description |
Wild fungal strains isolated from litter and soil were inoculated in test tubes containing a synthetic medium with 0.5% sodium carboxymethylcellulose (CMC) as sole carbon source. After the incubation time, cylindrical probes were taken from each tube using a borer and stained with Congo red 0.1% to reveal the remaining CMC. The relative cellulase activity was estimated by measuring the deepness of the clearing zone, and the growth by the limit of mycelial penetration in the medium. The ratio: hydrolysis zone/depth of growth, provides additional information to compare strains. A test using culture supernatants of liquid media growing strains was developed for enzyme assay using a modification of the cylinder-plate method. Petri dishes were filled with agar-CMC medium and cups were cut off with a steel punch. Such cups were filled with culture supernatants. After incubation, the plated were stained with Congo red, and the diameter of each cleared zone was recorded. Comparative studies of strains could be performed as well as quantitation of enzyme activity using a standard cellulase solution and computing the area of the cleared zones. This method may be useful when a large number of strains must be tested for cellulolytic activity or when conventional tests fail for assaying enzymatic induction in vitro. |
author |
Magnelli, Paula E. Martinez, Alicia Elba Mercuri, Oscar Alberto |
author_facet |
Magnelli, Paula E. Martinez, Alicia Elba Mercuri, Oscar Alberto |
author_sort |
Magnelli, Paula E. |
title |
Simple method for determining cellulolytic activity in fungi |
title_short |
Simple method for determining cellulolytic activity in fungi |
title_full |
Simple method for determining cellulolytic activity in fungi |
title_fullStr |
Simple method for determining cellulolytic activity in fungi |
title_full_unstemmed |
Simple method for determining cellulolytic activity in fungi |
title_sort |
simple method for determining cellulolytic activity in fungi |
publishDate |
1997 |
url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03257541_v29_n4_p210_Magnelli http://hdl.handle.net/20.500.12110/paper_03257541_v29_n4_p210_Magnelli |
work_keys_str_mv |
AT magnellipaulae simplemethodfordeterminingcellulolyticactivityinfungi AT martinezaliciaelba simplemethodfordeterminingcellulolyticactivityinfungi AT mercurioscaralberto simplemethodfordeterminingcellulolyticactivityinfungi |
_version_ |
1768543182062092288 |