Purification and characterization of two beta-glucosidases from the thermophilic fungus Thermoascus aurantiacus
Jazyk angličtina Země Spojené státy americké Médium print
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
12630320
DOI
10.1007/bf02818672
Knihovny.cz E-zdroje
- MeSH
- Ascomycota enzymologie MeSH
- beta-glukosidasa antagonisté a inhibitory chemie izolace a purifikace metabolismus MeSH
- chemická precipitace MeSH
- chromatografie iontoměničová MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- gelová chromatografie MeSH
- kinetika MeSH
- koncentrace vodíkových iontů MeSH
- molekulová hmotnost MeSH
- stříbro farmakologie MeSH
- ultrafiltrace MeSH
- vápník farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Brazílie MeSH
- Názvy látek
- beta-glukosidasa MeSH
- stříbro MeSH
- vápník MeSH
beta-Glucosidase from the fungus Thermoascus aurantiacus grown on semi-solid fermentation medium (using ground corncob as substrate) was partially purified in 5 steps--ultrafiltration, ethanol precipitation, gel filtration and 2 anion exchange chromatography runs, and characterized. After the first anion exchange chromatography, beta-glucosidase activity was eluted in 3 peaks (Gl-1, Gl-2, Gl-3). Only the Gl-2 and Gl-3 fractions were adsorbed on the gel matrix. Gl-2 and Gl-3 exhibited optimum pH at 4.5 and 4.0, respectively. The temperature optimum of both glucosidases was at 75-80 degrees C. The pH stability of Gl-2 (4.0-9.0) was higher than Gl-3 (5.5-8.5); both enzyme activities showed similar patterns of thermostability. Under conditions of denaturing gel chromatography the molar mass of Gl-2 and Gl-3 was 175 and 157 kDa, respectively. Using 4-nitrophenyl beta-D-glucopyranoside as substrate, Km values of 1.17 +/- 0.35 and 1.38 +/- 0.86 mmol/L were determined for Gl-2 and Gl-3, respectively. Both enzymes were inhibited by Ag+ and stimulated by Ca2+.
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