http://www.cnr.it/ontology/cnr/individuo/prodotto/ID168871
Comparison of extraction methods for recovery of extracellular B-glucosidase in two different forest soils. (Articolo in rivista)
- Type
- Label
- Comparison of extraction methods for recovery of extracellular B-glucosidase in two different forest soils. (Articolo in rivista) (literal)
- Anno
- 2008-01-01T00:00:00+01:00 (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi
- 10.1016/j.soilbio.2008.05.001 (literal)
- Alternative label
Masciandaro G., Macci C., Doni S., Maserti B.E.**, Calvo-Bado L.***, Ceccanti B., Wellington E.*** (2008)
Comparison of extraction methods for recovery of extracellular B-glucosidase in two different forest soils.
in Soil biology & biochemistry
(literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
- Masciandaro G., Macci C., Doni S., Maserti B.E.**, Calvo-Bado L.***, Ceccanti B., Wellington E.*** (literal)
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- Pagina fine
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume
- Rivista
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- ISI Web of Science (WOS) (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
- **Institute of Biophysics - CNR - Pisa (Italy)
***Dep. of Biological Sciences, Univ. of Warwick, Coventry (UK) (literal)
- Titolo
- Comparison of extraction methods for recovery of extracellular B-glucosidase in two different forest soils. (literal)
- Abstract
- A study was made of the efficiency of three different extractants, 0.1 M sodium pyrophosphate (pH 7),
67 mM phosphate buffer (pH 6) and 0.5 M potassium sulphate (pH 6.6), in recovering the protein
quantity and the b-glucosidase enzyme activity from two natural forest soils: (1) an Inceptisoil located in
Tuscany (Italy) in a mild Mediterranean climate, and (2) a Lithic Calcixeroll soil located in Murcia (southeast
of Spain) in a dry-semiarid climate. The pyrophosphate was used to determine the activity of
extracellular-humic-bound proteins, while the phosphate buffer and potassium sulphate were used to
extract dissolved extracellular proteins. The latter extractant, after chloroform fumigation, was also used
to measure total proteins in soil. A preliminary screening, using SDS-PAGE in one dimension, was also
carried out in order to optimize the separation condition of soil proteins extracted with different buffers.
To remove the interfering co-extracted substances (humic acid) a purification step using a column
packed with insoluble polyvinylpyrrolidone was performed. The highest b-glucosidase activity was
recovered in the pyrophosphate extract, thus confirming its capability of extracting humic-bound
b-glucosidase enzyme in a stable and active form. The extractants performed differently with the two soil
types and band patterns obtained with SDS-PAGE were extractant-specific, demonstrating that each was
selective for a particular class of proteins. Surprisingly, protein bands were also obtained using pyrophosphate,
in spite of the very dark extract colour due to the presence of humic substances. (literal)
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