T. Nikai et al., ISOLATION AND CHARACTERIZATION OF FIBRINOGENASE FROM CANDIDA-ALBICANSNH-1, International Journal of Biochemistry, 25(12), 1993, pp. 1815-1822
1. Fibrinogenase was isolated from Candida albicans NH-I by DEAE-Cellu
lose, Sephadex G-75 and Sephadex G-100 column chromatographies. 2. The
purified fibrinogenase gave a single band on disc polyacrylamide gel
electrophoresis, isoelectric focusing and sodium dodecyl sulfate polya
crylamide gel electrophoresis. 3. The enzyme preparation had a molecul
ar weight of 13,000, isoelectric point of pH 4.2 and possessed 117 ami
no acid residues. 4. The purified fibrinogenase possessed capillary pe
rmeability-increasing activity. 5. The enzyme hydrolyzed fibrinogen, c
asein, hide powder azure, azocoll hydrolytic activities and also hydro
lyzed the oxidized B chain of insulin. The cleavage sites in the oxidi
zed B chain of insulin were identified as Asp(3)-Glu(4), Glu(13)-Ala(1
4), Ala(14)-Leu(17), Tyr(16)-Leu(17), Arg(22)-Gly(23), Phe(25)Tyr(26)
and Tyr(26)-Thr(27). 6. Fibrinogenase activity of this preparation was
inhibited by alpha(2)-macroglobulin antithrombin-III, omicron-phenant
hroline, disodium ethylenediaminetetra acetic acid and dithiothreitol.