The rumen anaerobic fungi Orpinomyces joyonii A4 and Caecomyces communis JB1 were grown on microcrystalline cellulose (MC) and alfalfa hay. The cellular distribution of cellulases produced by these organisms was monitored. Fungal cultures were separated into extracellular, intracellular and cell wall fractions and assayed for endoglucanase (EG) and beta-glucosidase activity. In both fungal isolates, EG activity was the highest in the extracellular fraction regardless of the substrate used. The beta-glucosidase activity produced by O. joyonii was mainly found in the cell wall fraction. On the contrary, the same enzyme activity in C. communis predominated in the extracellular fraction. The polycentric isolate A4 more efficiently utilized both substrates, produced more short chain fatty acids (up to 31 mmol/l) and showed higher total levels of EG (2744 nmol glucose/h/ml) than the monocentric strain JB1. On the other hand, beta-glucosidase (9033 nmol glucose/h/ml) activity was the highest in cultures of C. communis grown on cellulose. In cultures of O. joyonii grown on MC, the production of yellow affinity substance (YAS) with similar properties compared with yellow substance from Clostridium thermocellum was observed. This compound increased the adsorption of fungal cellulases to MC the temperature and pH range tested.

Institute of Animal Physiology and Genetics Czech Academy of Sciences Prague Czech Republic

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Classical and molecular approaches as a powerful tool for the characterization of rumen polycentric fungi