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Glycoside hydrolases of family 75 include both eukaryotic (essentially fungal) and prokaryotic proteins. They have so far been characterized only from filamentous fungi. They are beta-1,4-chitosanases with endo-splitting activity [1, 2]. The analysis of the final product of hydrolysis of partially N-deacetylated chitosan by the GH75 chitosanase from Aspergillus fumigatus suggests that this enzyme cleaves preferentially GlcN-GlcN and GlcNAc-GlcN links in the polysaccharide chain [3].
Kinetics and Mechanism
Family GH46 enzymes are classified as inverting enzymes. This has been shown by 1H NMR for the enzyme from Aspergillus fumigatus using chitosan as substrate [3].
Catalytic Residues
A site-directed mutagenesis study performed on the enzyme from Fusarium solani (expressed as a recombinant protein in Saccharomyces cerevisiae) showed that Asp175 and Glu188 are essential for catalysis [4]. This was confirmed by a study on the chitosanase from Aspergillus fumigatus (expressed as a recombinant protein in Escherichia coli) showing that the corresponding residues Asp160 and Glu169 are essential for catalysis [3]. Both residues are strictly conserved among eukaryotic and prokaryotic GH75 family members.
Three-dimensional structures
No three-dimensional structure has been solved for this family.
Family Firsts
First primary structure determination
Chitosanase from Fusarium solani f. sp. phaseoli [5].
Shimosaka M, Nogawa M, Ohno Y, and Okazaki M. Chitosanase from the pathogenic fungus, Fusarium solani f.sp. phaseoli - purification and some properties. Biosci. Biotech. Biochem. 1993 57, 231-235. [Shimosaka1993]
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Shimosaka M, Kumehara M, Zhang X-Y, Nogawa M, and Okazaki M. Cloning and characterization of a chitosanase gene from the plant pathogenic fungus Fusarium solani. J. Ferment. Bioeng. 1996 82, 426-431. [Shimosaka1996]