HPAEC-PAD and Q-TOF-MS/MS analysis reveal a novel mode of action of endo-β-1,3(4)-d-glucanase Eng16A from coprinopsis cinerea on barley β-glucan.

We previously reported that an endo-β-1,3(4)-d-glucanase, Eng16A, from C. cinerea shows a higher degradation activity toward barley β-glucan than laminarin. HPAEC-PAD and Q-TOF-MS/MS analyses show that Eng16A-digestion products of barley β-glucan not only contain some oligosaccharides with (1 → 3)-β-linkage adjacent to the reducing end, which is consistent with β-1,3(4)-glucanase-digestion products, but also include some oligosaccharides containing (1 → 4)-β-linkage adjacent to the reducing end which is consistent with cellulase-digestion products.

Endo-β-1,3-glucanase digestion combined with the HPAEC-PAD-MS/MS analysis reveals the structural differences between two laminarins with different bioactivities.

To resolve the structure of laminarin, the recombinant endo-β-1,3-glucanase from Coprinopsis cinerea, which has specific activity on β-1,3 glycosidic bond and could hydrolyze the laminarin with complex structure, was used to hydrolyze laminarin. Then, the structures of enzyme-resistant oligosaccharides were quantitatively and qualitatively analysed by high-performance anion exchange chromatography coupled with mass spectrometry. The laminarin from Laminaria digitata contains 9.

The Modes of Action of ChiIII, a Chitinase from Mushroom Coprinopsis cinerea, Shift with Changes in the Length of GlcNAc Oligomers.

A putative class III endochitinase (ChiIII) was reported previously to be expressed dominantly in fruiting bodies of Coprinopsis cinerea, and its expression levels increased with the maturation of the fruiting bodies. This paper further reports that ChiIII is a novel chitinase with exo- and endoactivities. When the substrate was (GlcNAc)3-5, ChiIII exhibited exoactivity, releasing GlcNAc processively from the reducing end of (GlcNAc)3-5; when the substrate was (GlcNAc)6-7, the activity of ChiIII shifted to an endoacting enzyme, randomly splitting chitin oligosaccharides to various shorter oligosaccharides.

Comparative Study of Nonautolytic Mutant and Wild-Type Strains of Coprinopsis cinerea Supports an Important Role of Glucanases in Fruiting Body Autolysis.

Autolysis of Coprinopsis cinerea fruiting bodies affects its commercial value. In this study, a mutant of C. cinerea that exhibits pileus expansion without pileus autolysis was obtained using ultraviolet mutagenesis.