Current Genetics

, Volume 29, Issue 4, pp 404–409

Secretion of an enzymatically activeTrichoderma harzianum endochitinase bySaccharomyces cerevisiae

  • Henriette Draborg
  • Stephan Christgau
  • Torben Halkier
  • Grethe Rasmussen
  • Henrik Dalbøge
  • Sakari Kauminen
Original Paper

DOI: 10.1007/BF02208622

Cite this article as:
Draborg, H., Christgau, S., Halkier, T. et al. Curr Genet (1996) 29: 404. doi:10.1007/BF02208622

Abstract

A novel endochitinase agar-plate assay has been developed and used to identify 11 full-length cDNAs encoding endochitinase I (ENC I) from aTrichoderma harzianum cDNA library by expression in yeast. The 1473-bpchil cDNA encodes a 424-residue precursor protein including both a signal sequence and a propeptide. The deduced ENC I amino-acid sequence is homologous to other fungal and bacterial chitinases, and the enzyme cross-reacts with a polyclonal antiserum raised against chitinase A1 fromBacillus circulans. TheT. harzianum endochitinase I was secreted into the culture medium by the yeastSaccharomyces cerevisiae in a functionally active form. The purified recombinant enzyme had a molecular mass of 44 kDa, an isoelectric point of 6.3, a pH optimum of 7.0 and a temperature optimum of 20 °C.

Key words

Biocontrol Secretion Chitinase Expression cloning Saccharomyces cerevisiae Trichoderma harzianum 

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Henriette Draborg
    • 1
  • Stephan Christgau
    • 1
  • Torben Halkier
    • 1
  • Grethe Rasmussen
    • 1
  • Henrik Dalbøge
    • 1
  • Sakari Kauminen
    • 1
  1. 1.GeneSearchNovo Nordisk A/SBagsværdDenmark

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