Faculty of Food Technology and Biotechnology, University of Zagreb
Abstract
Seven species of basidiomycetes have been investigated for anti-complementary activity in hot water extracts and ethanol soluble fractions. Since Flammulina velutipes had the most potent activity, culture conditions for its mycelial growth were optimized to increase the production efficiency of anti-complementary exopolysaccharides. The optimal medium composition was (in g/L): galactose 15, sodium nitrate 5, glutamic acid 3, KH2PO4 2.5 and MgSO4·7H2O 0.6. Optimal production of anti-complementary activity was achieved at pH=3.5–5.5 and 25 °C. With these optimal medium and culture conditions, mycelial dry mass was maximized at 3.17 mg/mL after 6 days of cultivation in a 5-liter stirred-tank bioreactor, without pH control. The anti-complementary activity of exopolysaccharides increased sharply after 4 days of cultivation, and showed a high level at 5–6 days of cultivation. A periodate-labile carbohydrate moiety played a leading role in the anti-complementary activity exhibited by exopolysaccharide produced from F. velutipes. Results of tests on the anti-complementary activity in the absence of Ca²+ and immunoelectrophoresis indicated that the mode of complement activation by exopolysaccharide from F. velutipes is via both the classical and alternative pathways and that the activation degree is almost the same in each pathway.Istražena je antikomplementna aktivnost spojeva ekstrahiranih vrućom vodom i etanolom iz sedam vrsta gljiva stapčara. Optimirani su uvjeti uzgoja micelija gljive s najvećom aktivnosti, Flammulina velutipes, radi povećanja proizvodnje antikomplementnih egzopolisaharida. Optimalni sastav podloge bio je (u g/L): galaktoza 15, natrijev nitrat 5, glutamična kiselina 3, KH2PO4 2,5 i MgSO4·7H2O 0,6. Optimalna proizvodnja postignuta je pri pH=3,5-5,5 i 25 ºC. Pri tim uvjetima proizvedena je maksimalna količina suhe tvari od 3,17 mg/L nakon 6 dana uzgoja u bioreaktoru s miješalicom zapremnine 5 L, bez kontrole pH-vrijednosti. Antikomplementna aktivnost egzopolisaharida naglo se povećala nakon 4 dana, te je bila visoka nakon 5-6 dana uzgoja. Šećerni je ostatak, podložan djelovanju perjodata, glavni razlog antikomplementne aktivnosti egzopolisaharida gljive F. velutipes. Rezultati testiranja takve aktivnosti u odsutnosti Ca²+ iona i imunoelektroforeza upućuju na to da se komplementi egzopolisaharida iz F. velutipes aktiviraju klasičnim i alternativnim putem te da je stupanj aktivacije skoro jednak za oba puta
