Characterization of chitin and chitin synthase from the cellulosic cell wall fungusSaprolegnia monoi¨ca

1992; Academic Press; Volume: 16; Issue: 1 Linguagem: Inglês

10.1016/0147-5975(92)90037-r

ISSN

1878-4399

Autores

Vincent Bulone, H. Chanzy, Lucien Gay, Vincent Girard, M. FEVRE,

Tópico(s)

Microbial Natural Products and Biosynthesis

Resumo

The presence of chitin in hyphal cell walls and regenerating protoplast walls ofSaprolegnia monoi¨ca was demonstrated by biochemical and biophysical analyses. α-Chitin was characterized by X-ray diffraction, electron diffraction, and infrared spectroscopy. In hyphal cell walls, chitin appeared as small globular particles while cellulose, the other crystalline cell wall component, had a microfibrillar structure. Chitin synthesis was demonstrated in regenerating protoplasts by the incorporation of radioactiveN-acetylglucosamine into a KOH-insoluble product. Chitin synthase activity of cell-free extracts was particulate. This activity was stimulated by trypsin and inhibited by the competitive inhibitor polyoxin D (Ki 20 μM). The reaction product was insoluble in 1M KOH or 1M acetic acid and was hydrolyzed by chitinase into diacetylchitobiose. Fungal growth and cell wall chitin content were reduced when mycelia were grown in the presence of polyoxin D. However, hyphal morphology was not altered by the presence of the antibiotic indicating that chitin does not seem to play an important role in the morphogenesis ofSaprolegnia.

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