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Current Genetics

, Volume 29, Issue 5, pp 474–481 | Cite as

Expression of anErwinia pectate lyase in three species ofAspergillus

  • S. Bartling
  • J. P. T. W. van den Hombergh
  • O. Olsen
  • D. von Wettstein
  • J. Visser
Original Paper
  • 44 Downloads

Abstract

Transgenic filamentous fungi of the speciesAspergillus niger, A. nidulans andA. awamori expressing and secretingErwinia carotovora subsp.atroseptica pectate lyase 3 (PL3) were generated. Correct processing of the pre-enzyme was achieved using theA. niger pectin lyase A (PEL A) signal peptide. With the prepro-peptide ofA. niger polygalacturonase II, secreted enzymes still possessed the 6- aa pro-sequence, indicating the importance of the conformation of the precursor protein for correct cleavage of the signal sequence. PL3 expression was markedly increased in media optimized for limited protease activity, and reached 0.4, 0.8 and 2.0 mg/l for expression inA. niger, A. awamori andA. nidulans, respectively. Glycans attached to the PL3 enzymes exhibited species-specific differences, and an increase of molecular mass coincided with reduced specific activities of the enzymes.

Keywords

Proteolytic degradation Expression Processing Glycosylation 

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Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • S. Bartling
    • 1
  • J. P. T. W. van den Hombergh
    • 2
  • O. Olsen
    • 1
  • D. von Wettstein
    • 1
  • J. Visser
    • 2
  1. 1.Department of PhysiologyCarlsberg LaboratoryCopenhagenDenmark
  2. 2.Section Molecular Genetics of Industrial MicroorganismsWageningen Agricultural UniversityWageningenThe Netherlands

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