Molecular and General Genetics MGG

, Volume 263, Issue 2, pp 281–286

Cloning and characterization of oah, the gene encoding oxaloacetate hydrolase in Aspergillus niger

  • H. Pedersen
  • C. Hjort
  • J. Nielsen
ORIGINAL PAPER

DOI: 10.1007/s004380051169

Cite this article as:
Pedersen, H., Hjort, C. & Nielsen, J. Mol Gen Genet (2000) 263: 281. doi:10.1007/s004380051169
  • 169 Downloads

Abstract

The enzyme oxaloacetate hydrolase (EC 3.7.1.1), which is involved in oxalate formation, was purified from Aspergillus niger. The native enzyme has a molecular mass of 360–440 kDa, and the denatured enzyme has a molecular mass of 39 kDa, as determined by gel electrophoresis. Enzyme activity is maximal at pH 7.0 and 45 °C. The fraction containing the enzyme activity contained at least five proteins. The N-terminal amino acid sequences of four of these proteins were determined. The amino acid sequences were aligned with EST sequences from A. niger, and an EST sequence that showed 100% identity to all four sequences was identified. Using this EST sequence the gene encoding oxaloacetate hydrolase (oah) was cloned by inverse PCR. It consists of an ORF of 1227 bp with two introns of 92 and 112 bp, respectively. The gene encodes a protein of 341 amino acids with a molecular mass of 37 kDa. Under the growth conditions tested, the highest oah expression was found for growth on acetate as carbon source. The gene was expressed only at pH values higher than 4.0.

Key words Oxalate Aspergillus Oxaloacetate hydrolase Oxalacetase 

Copyright information

© Springer-Verlag Berlin Heidelberg 2000

Authors and Affiliations

  • H. Pedersen
    • 1
  • C. Hjort
    • 2
  • J. Nielsen
    • 1
  1. 1.Center for Process Biotechnology, Department of Biotechnology, Building 227, Technical University of Denmark, 2800 Lyngby, Denmark E-mail: jn@ibt.dtu.dk Tel.: +45-45-252696; Fax: +45-45-884148DK
  2. 2.Novo Nordisk A/S, DK-2880 Bagsvaerd 2, DenmarkDK

Personalised recommendations