| Abstract: | Extraction of free amino acids from fungal spores (Phycornyces blakesleeanus) Because extraction conditions are crucial for the reliability of biochemical analysis of fungal spore materia1, comparative investigations on methods were conducted, using the amount of free amino acids as a parameter of exhaustive extraction. It was found that optimal conditions are: Extraction in a Soxhlet apparatus with methanol as extracting medium (for 8 hours under reduced pressure and 40°C boiling temperature) or treatment with ice-cold perchloric acid (10 % for 30 minutes) or trichloroacetic acid (10 % for 30 minutes). A pretreatment of the material by desintegration was not necessary. Extraction with hot water (70 or l00T) or with ethanol, propanol, butanol, 3,3 %perchloric or trichloroacetic acid proved to be inappropriate. All of the common amino acids were detected in the extracts except proline and phenylalanine. A few amino acids make up the majority of the pool (glutamic acid, glutamine, aspartic acid, asparagine, alanine and y-butyric acid). These amino acids comprised about 80% of the total intracellular pool (ca. 12000 moles/l00mg spores) and account for nearly 2% of the spore dry weight. Sacrosine, probably being present in considerable amounts, could not be determined quantitatively. |
