The first characterized asparaginase from a basidiomycete, Flammulina velutipes

Research output: Contribution to journalArticleResearchpeer review

Authors

  • Nadine Eisele
  • Diana Linke
  • Katrin Bitzer
  • Shukry Na'amnieh
  • Manfred Nimtz
  • Ralf G. Berger

Research Organisations

External Research Organisations

  • Helmholtz Centre for Infection Research (HZI)
  • X-Zyme Biotechnology GmbH
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Details

Original languageEnglish
Pages (from-to)3316-3321
Number of pages6
JournalBioresource technology
Volume102
Issue number3
Publication statusPublished - 25 Oct 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.

Keywords

    Asparaginase, Basidiomycete, Escherichia coli, Flammulina velutipes, Heterologous expression

ASJC Scopus subject areas

Sustainable Development Goals

Cite this

The first characterized asparaginase from a basidiomycete, Flammulina velutipes. / Eisele, Nadine; Linke, Diana; Bitzer, Katrin et al.
In: Bioresource technology, Vol. 102, No. 3, 25.10.2010, p. 3316-3321.

Research output: Contribution to journalArticleResearchpeer review

Eisele, N, Linke, D, Bitzer, K, Na'amnieh, S, Nimtz, M & Berger, RG 2010, 'The first characterized asparaginase from a basidiomycete, Flammulina velutipes', Bioresource technology, vol. 102, no. 3, pp. 3316-3321. https://doi.org/10.1016/j.biortech.2010.10.098
Eisele, N., Linke, D., Bitzer, K., Na'amnieh, S., Nimtz, M., & Berger, R. G. (2010). The first characterized asparaginase from a basidiomycete, Flammulina velutipes. Bioresource technology, 102(3), 3316-3321. https://doi.org/10.1016/j.biortech.2010.10.098
Eisele N, Linke D, Bitzer K, Na'amnieh S, Nimtz M, Berger RG. The first characterized asparaginase from a basidiomycete, Flammulina velutipes. Bioresource technology. 2010 Oct 25;102(3):3316-3321. doi: 10.1016/j.biortech.2010.10.098
Eisele, Nadine ; Linke, Diana ; Bitzer, Katrin et al. / The first characterized asparaginase from a basidiomycete, Flammulina velutipes. In: Bioresource technology. 2010 ; Vol. 102, No. 3. pp. 3316-3321.
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AU - Eisele, Nadine

AU - Linke, Diana

AU - Bitzer, Katrin

AU - Na'amnieh, Shukry

AU - Nimtz, Manfred

AU - Berger, Ralf G.

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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.

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