1887

Abstract

The gene encoding a major, inducible 45 kDa chitinase of was cloned and analysis of the deduced amino acid sequence identified a chitinase of the fungal/bacterial class which was designated ChiB1. Recombinant ChiB1, expressed in , was shown to function by a retaining mechanism of action. That is, the -conformation of the chitin substrate linkage was preserved in the product in a manner typical of family 18 chitinases. Cleavage patterns with the -acetylglucosamine (GlcNAc) oligosaccharide substrates GlcNAc, GlcNAc and GlcNAc indicated that the predominant reaction involved hydrolysis of GlcNAc from the non-reducing end of each substrate. Products of transglycosylation were also identified in each incubation. Following disruption of by gene replacement, growth and morphology of disruptants and of the wild-type strain were essentially identical. However, during the autolytic phase of batch cultures the level of chitinase activity in culture filtrate from a disruptant was much lower than the activity from the wild-type. The search for chitinases with morphogenetic roles in filamentous fungi should perhaps focus on chitinases of the fungal/plant class although such an investigation will be complicated by the identification of at least 11 putative active site domains for family 18 chitinases in the TIGR database (http://www.tigr.org/).

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2003-10-01
2019-10-15
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