A systematic study on autolysis of the cell walls of fungi has been made on Neurospora crassa, Botrytis cinerea, Polystictus versicolor, Aspergillus nidulans, Schizophyllum commune, Aspergillus niger, and Mucor mucedo. During autolysis each fungus produces the necessary lytic enzymes for its autodegradation. From autolyzed cultures of each fungus enzymatic precipitates were obtained. The degree of lysis of the cell walls, obtained from non-autolyzed mycelia, was studied by incubating these cell walls with and without a supply of their own lytic enzymes. The degree of lysis increased with the incubation time and generally was higher with a supply of lytic enzymes. Cell walls from mycelia of different ages were obtained. A higher degree of lysis was always found, in young cell walls than in older cell walls, when exogenous lytic enzymes were present. In all the fungi studied, there is lysis of the cell walls during autolysis. This is confirmed by the change of the cell wall structure as well as by the degree of lysis reached by the cell wall and the release of substances, principally glucose and N-acetylglucosamine in the medium.

The degrees of autolysis attained by five different genera of filamentous fungi during an incubation period of 60 days, under the same culture conditions were: 87.3% for Penicillium oxalicum; 65.9% for Neurospora crassa; 62.7% for Polystictus versicolor; 51.7% for Aspergillus niger and 23.5% for Nectria galligena. N. crassa, A. niger and P. versicolor reached the end of the autolysis during this incubation period (60 days), whereas P. oxalicum and N. galligena did not. The excretion of the lytic enzymes beta-N-acetylglucosaminidase, beta -1-3 glucanase, chitinase, invertase and acid phosphatase into the culture medium during growth and autolysis was investigated. The excretion of these enzymes was consistent with the degree of autolysis reached, the maximum excretion belonging to P. oxalicum and the minimum to N. galligena. The N. crassa invertase was excreted into the culture liquid at levels very much higher than the other enzymes studied, and at levels very much higher than the invertases excreted by the other fungi.

Cultures of Penicillium oxalicum growing on a denned medium supplemented with yeast extract reached the onset of autolysis after 3 days at 25 °C. Thenceforth, autolysis was progressive and eventual reductions in dry weight of 96% were recorded by day 47. The pH of the medium fluctuated between 4.0 during the exponential phase of growth and 9.0 during autolysis. Electron microscopy of autolyzing cultures revealed a progressive loss of cytoplasmic ultrastructure. Digestion of the cell walls, with a rapid hydrolysis of the three external layers and a low hydrolysis of the two inner layers, was accompanied by deep pitting and by loss of the distinct five-layered structure. A lytic enzyme complex was obtained from the filtrates of extensively autolyzed cultures. It was rich in (1 → 3)-β-glucanase and other enzymes active against a range of fungal cell wall and storage polysaccharides. This enzyme complex degraded extensively isolated cell walls of P. oxalicum and three other Ascomycetes but had less effect on walls isolated from Mucor mucedo or Schizophyllum commune. In the case of P. oxalicum, cell walls harvested from young cultures were more readily digested than were the walls from older cultures.

~~~~~~The synthesis of 1,3-P-glucanase, P-N-acetylglucosaminidase and total protein was studied during autolysis of Neurospora crassa. Evidence for de novo protein synthesis during autolysis was obtained from the incorporation of [3H]tryptophan and [3H]phenylalanine into the protein pool and from the inhibitory effects of cycloheximide and puromycin. Glucose accumulated in the growth medium during autoly sis and N-acetylglucosamine and two unidentified oligosaccharides were detected towards the end of the 46 d autolysis period.During autolysis, 1,3-P-glucanase accumulated in the medium to tenfold greater activities than P-N-acetylglucosaminidase. The former enzyme had a pH optimum of 5 . 5 and a K , of 0.45 mM. It was inhibited by excess substrate but not by excess glucose.

No abstract