The Chemistry and Molecular Architecture of the Cell Walls of Penicillium chrysogenum
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Abstract Treatment of 14C-uniformly labeled cell walls of Penicillium chrysogenum with chitobiase-containing chitinase resulted in 73% solubilization. The solubilized components were isolated, identified, and determined quantitatively. N-Acetylglucosamine (GlcNAc) and N,N'-diacetylchitobiose ((GlcNAc)2) accounted for 42% of the cell wall. Since kinetic analysis showed that GlcNAc arose from (GlcNAc)2, at least 42% of the cell wall contains GlcNAc in β-d-(1 → 4)-glycosidic linkage. When the same experiment was performed with a chitobiase-free chitinase preparation nearly identical results were obtained, except, as expected, essentially all of the GlcNAc was present as (GlcNAc)2. Small amounts of glucose and laminaritriose were also released. In addition, a nondialyzable component, Fraction 5A, accounting for 14% of the wall, was isolated. This fraction, devoid of GlcNAc, contains 20 times the amount of galactose and 2 times the amount of mannose with respect to glucose found in cell wall hydrolysates. An additional 13% of the wall was solubilized when the cell wall residue, following treatment with chitinase, was treated with chitinase-free β-d-(1 → 3)-glucanase. Therefore, 86% of the wall was solubilized when treated first with chitinase, then with β-d-(1 → 3)-glucanase. Initial treatment of cell walls with β-d-(1 → 3)-glucanase resulted in 48% solubilization. From an analysis of the solubilized components the conclusion was reached that at least 40% of the wall contains glucose in β-d-(1 → 3)-glycosidic linkage. An additional 44% of the wall was solubilized when the residue, following treatment with β-d-(1 → 3)-glucanase, was treated with chitinase. Thus, 92% of the wall was solubilized when treated first with β-d-(1 → 3)-glucanase, then with chitinase. The observations that β-d-(1 → 3)-glucanase-free chitinase released in addition to GlcNAc and (GlcNAc)2 also glucose, laminaritriose, and a fraction containing mannose, galactose, and glucose (Fraction 5A) and that treatment of this fraction with β-d-(1 → 3)-glucanase released glucose and laminaribiose suggest that glucose may exist in cell walls in one or more forms other than the major glucan. From the available data it is not clear whether Fraction 5A contains one or several polymers. An analysis of the monomeric cell wall constituents released by acid hydrolysis is in excellent agreement with the results obtained by enzymatic digestion. These results showed the cell wall to contain about 48% glucosamine and about 52% neutral hexoses, which consisted of glucose, galactose, and mannose in the molar proportion of 10:1:2. Thus, if 40% of the wall is glucose, then galactose and mannose account for 4% and 8% of the wall, respectively.Keywords:
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