Applied Microbiology and Biotechnology

, Volume 44, Issue 1–2, pp 147–156

Efficient expression and secretion of Aspergillus niger RH5344 polygalacturonase in Saccharomyces cerevisiae

  • C. Lang
  • A. C. Looman
Applied Genetics and Regulation

DOI: 10.1007/BF00164494

Cite this article as:
Lang, C. & Looman, A.C. Appl Microbiol Biotechnol (1995) 44: 147. doi:10.1007/BF00164494

Abstract

An Aspergillus niger endopolygalacturonase (EC 3.2.1.15) cDNA was expressed in the yeast Saccharomyces cerevisiae. Secretion of the protein into the growth medium was efficiently directed by the fungal leader sequence, and processing occurred at the same site as in Aspergillus. The expression level was significantly enhanced by using a “short” version of the yeast ADHI promoter. An additional increase in the yield of heterologous protein was due to a higher plasmid stability and a rise in plasmid copy number. This was achieved by deleting most of the bacterial sequences from the expression vector. The yeast-derived enzyme showed the same enzymatic and biochemical properties as the fungal polygalacturonase, such as substrate specificity, pH and temperature optima and pI value. The yeast-derived enzyme, however, showed a higher degree of glycosylation and exhibited a more pronounced temperature stability than the fungal enzyme.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • C. Lang
    • 1
  • A. C. Looman
    • 1
  1. 1.Institut für Gärungsgewerbe und BiotechnologieAbt. Biotechnologie HülsBerlinGermany
  2. 2.Department of Food Sciences and Biotechnology, Institute of Microbiology and GeneticsTechnische Universität BerlinBerlinGermany
  3. 3.Department of NeonatologyHumboldt-Universität Berlin, University Hospital CharitéBerlinGermany

Personalised recommendations