Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 3316-3321 |
| Seitenumfang | 6 |
| Fachzeitschrift | Bioresource technology |
| Jahrgang | 102 |
| Ausgabenummer | 3 |
| Publikationsstatus | Veröffentlicht - 25 Okt. 2010 |
Abstract
Flammulina velutipes enjoys high popularity as an edible mushroom in Asian cuisines. Investigating the secretion of peptidases in nutrient media enriched with gluten, an enzyme was noticed that catalyzed the deamidation of l-asparagine and l-glutamine. The enzyme was purified to electrophoretic homogeneity by foaming and SEC. PAGE analysis revealed a protein of about 85 kDa with 13 kDa subunits indicating a hexameric protein. Degenerated primers were deduced from peptide fragments and the complete coding sequence of 372 bp was determined. The gene of Flammulina velutipes asparaginase (FvNase) over expressed in E. coli achieved an l-asparagine-hydrolyzing activity of 16. U/mL in crude extract, which was five times higher than its l-glutamine-hydrolyzing ability. The enzyme showed a pH-optimum at pH 7, remarkable tolerance towards elevated temperature and sodium chloride concentration in both the native and recombinant form, and no significant homology to any conserved domains of published asparaginases or glutaminases.
ASJC Scopus Sachgebiete
- Chemische Verfahrenstechnik (insg.)
- Bioengineering
- Umweltwissenschaften (insg.)
- Environmental engineering
- Energie (insg.)
- Erneuerbare Energien, Nachhaltigkeit und Umwelt
- Umweltwissenschaften (insg.)
- Abfallwirtschaft und -entsorgung
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in: Bioresource technology, Jahrgang 102, Nr. 3, 25.10.2010, S. 3316-3321.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - The first characterized asparaginase from a basidiomycete, Flammulina velutipes
AU - Eisele, Nadine
AU - Linke, Diana
AU - Bitzer, Katrin
AU - Na'amnieh, Shukry
AU - Nimtz, Manfred
AU - Berger, Ralf G.
PY - 2010/10/25
Y1 - 2010/10/25
N2 - Flammulina velutipes enjoys high popularity as an edible mushroom in Asian cuisines. Investigating the secretion of peptidases in nutrient media enriched with gluten, an enzyme was noticed that catalyzed the deamidation of l-asparagine and l-glutamine. The enzyme was purified to electrophoretic homogeneity by foaming and SEC. PAGE analysis revealed a protein of about 85 kDa with 13 kDa subunits indicating a hexameric protein. Degenerated primers were deduced from peptide fragments and the complete coding sequence of 372 bp was determined. The gene of Flammulina velutipes asparaginase (FvNase) over expressed in E. coli achieved an l-asparagine-hydrolyzing activity of 16. U/mL in crude extract, which was five times higher than its l-glutamine-hydrolyzing ability. The enzyme showed a pH-optimum at pH 7, remarkable tolerance towards elevated temperature and sodium chloride concentration in both the native and recombinant form, and no significant homology to any conserved domains of published asparaginases or glutaminases.
AB - Flammulina velutipes enjoys high popularity as an edible mushroom in Asian cuisines. Investigating the secretion of peptidases in nutrient media enriched with gluten, an enzyme was noticed that catalyzed the deamidation of l-asparagine and l-glutamine. The enzyme was purified to electrophoretic homogeneity by foaming and SEC. PAGE analysis revealed a protein of about 85 kDa with 13 kDa subunits indicating a hexameric protein. Degenerated primers were deduced from peptide fragments and the complete coding sequence of 372 bp was determined. The gene of Flammulina velutipes asparaginase (FvNase) over expressed in E. coli achieved an l-asparagine-hydrolyzing activity of 16. U/mL in crude extract, which was five times higher than its l-glutamine-hydrolyzing ability. The enzyme showed a pH-optimum at pH 7, remarkable tolerance towards elevated temperature and sodium chloride concentration in both the native and recombinant form, and no significant homology to any conserved domains of published asparaginases or glutaminases.
KW - Asparaginase
KW - Basidiomycete
KW - Escherichia coli
KW - Flammulina velutipes
KW - Heterologous expression
UR - http://www.scopus.com/inward/record.url?scp=78650808076&partnerID=8YFLogxK
U2 - 10.1016/j.biortech.2010.10.098
DO - 10.1016/j.biortech.2010.10.098
M3 - Article
C2 - 21075625
AN - SCOPUS:78650808076
VL - 102
SP - 3316
EP - 3321
JO - Bioresource technology
JF - Bioresource technology
SN - 0960-8524
IS - 3
ER -