ISSN:
1432-0983
Keywords:
Biocontrol
;
Secretion
;
Chitinase
;
Expression cloning
;
Saccharomyces cerevisiae
;
Trichoderma harzianum
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF02208622
