PMID: 8598062Mar 1, 1996Paper

Secretion of an enzymatically active Trichoderma harzianum endochitinase by Saccharomyces cerevisiae

Current Genetics
H DraborgS Kauppinen

Abstract

A novel endochitinase agar-plate assay has been developed and used to identify 11 full-length cDNAs encoding endochitinase I (ENCI) from a Trichoderma harzianum cDNA library by expression in yeast. The 1473-bp chi1 cDNA encodes a 424-residue precursor protein including both a signal sequence and a propeptide. The deduced ENCI amino-acid sequence is homologous to other fungal and bacterial chitinases, and the enzyme cross-reacts with a polyclonal antiserum raised against chitinase A1 from Bacillus circulans. The T. harzianum endochitinase I was secreted into the culture medium by the yeast Saccharomyces 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 degrees C.

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Citations

Nov 7, 2006·Journal of Microbiological Methods·Sébastien Massart, Haissam M Jijakli
Jun 24, 1999·Acta Microbiologica Et Immunologica Hungarica·L Manczinger
Jun 9, 2006·Mycopathologia·Li Duo-Chuan
Aug 19, 2007·Protein Expression and Purification·Yanhua FanYan Pei

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