the catalytic activity and stereoselectivity of ficin are significantly influenced by reaction media. Best yield of 34% with a poor enantioselectivity of 29% in THF. Best enantioselectivity of 57% with a yield of 28% in MeCN. In tested polar solvents including i-PrOH, DMSO, and DMF, ficin shows lower activityand enantioselectivity
the inhibitor increases the noncovalent reversible binding affinity of ficin towards the maleimides without affecting the nucleophilic reactivity of the catalytically essential thiol groups which are alkylated in the inactivation reaction
ficin E is not affected by Mg2+, Ca2+ and Mn2+ at a concentration up to 10 mM. It is unaffected by synthetic (Pefabloc SC, benzamidine) and by natural proteinaceous (aprotinin) serine proteases inhibitors, by aspartic proteases inhibitors (pepstatin A) and by metallo-proteases inhibitors (EDTA, EGTA)
Immunocytochemical assay for estrogen receptor with monoclonal antibody D753P gamma in routinely processed formaldehyde-fixed breast tissue. Comparison with frozen section assay and with monoclonal antibody H222.
ficin looses 37% of its activity at 70Â°C after 10 min of incubation, and in the presence of co-solvents, the activity is protected up to 91%, maximum protection is shown by trehalose and followed by sorbitol, sucrose, and xylitol
ficin is threatened by two main factors, autolysis and aggregation. Autolysis of ficin in the absence of inhibitor is very large but in the presence of iodoacetamide it is completely inhibited. However ficin aggregation is greatly induced. This is because of beta-sheets increasing in the second structure, structure opening, ficin molecules adhesion to each other and molecules sedimentation. Iodoacetamide inhibits ficin activity irreversibility and aggregates large amount of protein. Therefore, iodoacetamide is not recommended for autolysis inhibition because of large aggregation. Tetrathionate has both activator and inhibitor effect on ficin autolysis at very low concentrations and higher concentrations respectively. Tetrathionate has a few side effects on structure opening, thermal stability decreasing and aromatic residues exiting. A tetrathionate inhibitory effect is reversible on ficin by dialyzing.
the enzyme activity decreases significantly at low concentration of urea before unfolding of the enzyme molecule (about 50% residual activity at 2 M, about 20% residual activity at 4 M, no activity at 6 M)
further purification of a commercial latex preparation by cation exchange chromatography, and 2,2'-dipyridyl disulfide-glutathione or aminophenylmercuric acetate affinity chromatography, or by Gly-Phe-NH-methylcyanide affinity chromatography with elution by 2,2'-dipyridyl disulfide
prolonged stability of ficin at low pH values in comparison to papain can be of importance for industrial processes that run in low pH conditions such as chill haze prevention during winemaking which prompted us to check long term stability of ficin and papain at low pH and in the presence of ethanol
kinetically controlled formation of peptide bonds by coupling the ester substrates benzyloxycarbonyl-Ala methyl ester and benzyloxycarbonyl-Gly methyl ester with L-Ala, D-Ala, L-Gln, D-Gln and L-Cys(acetamidomethyl) respectively