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.
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Communicated by A. Hinnen
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Draborg, H., Christgau, S., Halkier, T. et al. Secretion of an enzymatically activeTrichoderma harzianum endochitinase bySaccharomyces cerevisiae . Curr Genet 29, 404–409 (1996). https://doi.org/10.1007/BF02208622
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DOI: https://doi.org/10.1007/BF02208622