Affordable Access

Publisher Website

N1,N12-Diacetylspermine oxidase fromDebaryomyces hanseniiT-42: Purification, characterization, molecular cloning and gene expression

Authors
Journal
Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics
1570-9639
Publisher
Elsevier
Publication Date
Volume
1774
Issue
11
Identifiers
DOI: 10.1016/j.bbapap.2007.08.010
Keywords
  • Diacetylspermine Oxidase
  • Debaryomyces Hansenii
  • Substrate Specificity
  • Cloning
  • Heterologous Expression
Disciplines
  • Biology

Abstract

Abstract An FAD-dependent N 1, N 12-diacetylspermine oxidase (DASpmOX), which seems suitable for enzymatic determination of the tumor marker N 1, N 12-diacetylspermine (DASpm), was isolated from Debaryomyces hansenii T-42. DASpmOX exhibited the most excellent specificity toward DASpm among all polyamine oxidases found to date, and the specificity for DASpm could be raised by adjusting the pH of the buffer and adding TritonX-100. In potassium phosphate (pH 7.0) with 0.3% TritonX-100, this enzyme did not have any detectable activity toward free polyamines, and the reaction rate of N 1, N 8-diacetylspermidine, N 1-acetylspermine, N 1-acetylspermidine, and N 8-acetylspermidine was only 19%, 7.8%, 7.8%, and 1.0% of that of DASpm, respectively. The gene encoding DASpmOX was cloned and expressed in Escherichia coli. The apparent k cat and K m values of recombinant enzyme for DASpm were found to be 158 s − 1 and 3.1 × 10 − 4 M under the conditions described above, respectively.

There are no comments yet on this publication. Be the first to share your thoughts.