TY - JOUR

T1 - Purification, characterisation and cDNA sequencing of pyruvate decarboxylase from Zygosaccharomyces bisporus

AU - Neuser, Frauke

AU - Zorn, Holger

AU - Richter, Ulla

AU - Berger, Ralf G.

N1 - Funding information: Frauke Neuser and Ulla Richter were financially supported by the Deutsche Forschungsgemeinschaft.

PY - 2000/4

Y1 - 2000/4

N2 - Cells of the wild-type yeast strain Zygosaccharomyces bisporus CBS 702 form α-hydroxy ketones from aromatic amino acid precursors during fermentation. Pyruvate decarboxylase (PDC, E.C. 4.1.1.1), the key enzyme of this biotransformation catalysing the non-oxidative decarboxylation of pyruvate and other 2oxo-acids, was purified and characterised. The active enzyme is homotetrameric (α4) with a molecular mass of about 244 kDa. Activation of PDC by its substrate pyruvate results in a sigmoidal dependence of the reaction rate from substrate concentration (apparent K(m) value 1.73 mM; Hill coefficient 2.10). A cDNA library was screened using a PCR-based procedure, and a 1856 bp cDNA of PDC was identified and sequenced. The cDNA encodes a polypeptide of 563 amino acid residues (monomeric unit). Sequence alignments demonstrate high homologies (> 80%) to PDC genes from Saccharomyces cerevisiae, Kluyveromyces lactis and Kluyveromyces marxianus.

AB - Cells of the wild-type yeast strain Zygosaccharomyces bisporus CBS 702 form α-hydroxy ketones from aromatic amino acid precursors during fermentation. Pyruvate decarboxylase (PDC, E.C. 4.1.1.1), the key enzyme of this biotransformation catalysing the non-oxidative decarboxylation of pyruvate and other 2oxo-acids, was purified and characterised. The active enzyme is homotetrameric (α4) with a molecular mass of about 244 kDa. Activation of PDC by its substrate pyruvate results in a sigmoidal dependence of the reaction rate from substrate concentration (apparent K(m) value 1.73 mM; Hill coefficient 2.10). A cDNA library was screened using a PCR-based procedure, and a 1856 bp cDNA of PDC was identified and sequenced. The cDNA encodes a polypeptide of 563 amino acid residues (monomeric unit). Sequence alignments demonstrate high homologies (> 80%) to PDC genes from Saccharomyces cerevisiae, Kluyveromyces lactis and Kluyveromyces marxianus.

KW - Acyloin formation

KW - Enzyme purification

KW - Non-conventional yeast

KW - Sequence alignment

KW - α-Hydroxy ketones

UR - http://www.scopus.com/inward/record.url?scp=0034056364&partnerID=8YFLogxK

U2 - 10.1515/BC.2000.046

DO - 10.1515/BC.2000.046

M3 - Article

C2 - 10839465

AN - SCOPUS:0034056364

VL - 381

SP - 349

EP - 353

JO - Biological chemistry

JF - Biological chemistry

SN - 1431-6730

IS - 4

ER -