FUMARIC-ACID, ANTI-THROMBIN SUBSTANCE FROM RHIZOPUS-JAVANICUS

An anti-thrombin substance (M2) was isolated from a culture broth of R hizopus javanicus. Accumulation of M2 reached a maximum peak after 14 to 15 days of incubation and then decreased. The yield of M2 was 500 m g from 1 L of culture broth. M2 inhibited thrombin activity, and its 5 0% inhibition concentration in a reaction mixture containing 50 mu L o f 12.5 NIH unit/mL thrombin and 200 mu L of 0.33% bovine fibrinogen wa s 63 mu M. M2 had a specific activity for thrombin, but it was less re sponsive to plasmin, tissue-type plasminogen activator, urokinase, pla sma kallikrein and glandular kallikrein. The structure of M2 was ident ified as fumaric acid by elementary analysis, FAB/MS, H-1-NMR, C-13-NM R and IR spectra.