Fumaric acid, anti-thrombin substance from Rhizopus javanicus

Area of Science:

• Biochemistry
• Microbiology
• Enzymology

Background:

• Thrombin plays a critical role in blood coagulation.
• The search for novel anticoagulants is ongoing for therapeutic applications.
• Rhizopus javanicus is a known source of bioactive compounds.

Purpose of the Study:

• To isolate and characterize an anti-thrombin substance from Rhizopus javanicus culture broth.
• To determine the inhibitory activity and specificity of the isolated substance against thrombin.
• To elucidate the chemical structure of the anti-thrombin substance.

Main Methods:

• Isolation of the anti-thrombin substance (M2) from Rhizopus javanicus culture broth.
• Measurement of M2 accumulation over 14-15 days of incubation.
• Determination of M2 yield (500 mg/L).
• Enzyme inhibition assays to assess M2's effect on thrombin, plasmin, and other proteases.
• Structural elucidation using elementary analysis, FAB/MS, 1H-NMR, 13C-NMR, and IR spectroscopy.

Main Results:

• An anti-thrombin substance, M2, was isolated and reached maximum accumulation after 14-15 days.
• The yield of M2 was 500 mg/L of culture broth.
• M2 demonstrated specific inhibitory activity against thrombin (50% inhibition at 63 microM) with limited effect on other proteases.
• The chemical structure of M2 was identified as fumaric acid.

Conclusions:

• Rhizopus javanicus produces an anti-thrombin substance identified as fumaric acid.
• Fumaric acid exhibits specific inhibitory activity towards thrombin.
• This finding suggests fumaric acid as a potential lead compound for anticoagulant development.