ABSTRACT
Three enzymes with milk-clotting activity have been isolated from the fruiting bodies of Pleurotus ostreatus (Fr.) Kumm.) by (NH4)2SO4 precipitation, gel chromatography on Sephadex G75, and ion exchange chromatography on carboxymethylcellulose (CMC). Isoelectric points of the enzymes, as determined by isoelectrofocusing, equaled 4.2, 6.7, and 8.8. Inhibition analysis showed that the enzymes with isoelectric points of 4.2 and 6.7 belong to the class of metal-dependent proteinases, while the enzyme with the isoelectric point of 8.8 belongs to the serine protease class.
Subject(s)
Fruiting Bodies, Fungal/enzymology , Fungal Proteins/isolation & purification , Metalloproteases/isolation & purification , Milk , Pleurotus/enzymology , Serine Proteases/isolation & purification , Animals , Cattle , Fungal Proteins/chemistry , Isoelectric Point , Metalloproteases/chemistry , Serine Proteases/chemistryABSTRACT
The activity of trypsin-like proteinases and trypsin inhibitors was measured in fruiting bodies of various species of basidial fungi (Basidiomycetes). Fruiting bodies of all fungi contained these enzymes, with the exceptions of polypore (Coriolus versicolor (Fr.) Karst) and hedgehog fungus (Hericium erinaceus (Fr.) Quel), belonging to the families Polyporaceae and Hericiaceae, respectively, in which the enzyme activities were barely detectable. The activity of trypsin-like proteinases was the highest in fruiting bodies of Boletaceae and Agaricaceae. Fruiting bodies of all fungi contained trypsin inhibitors. The highest activity of trypsin inhibitors was detected in basidiomycetes of the families Boletaceae, Agaricaceae, and Pleurotaceae, including Boletus castanus (Fr.) Karst, orange-cap boletus (Leccinum aurantiacum (Fr.) Sing), and brown-cap boletus (Leccinum melanum (Fr.) Karst).
Subject(s)
Basidiomycota/enzymology , Fruiting Bodies, Fungal/enzymology , Peptide Hydrolases/metabolism , Trypsin Inhibitors/metabolism , Trypsin/metabolismABSTRACT
Three trypsin isoforms (designated as T1, T2, and T3), three chymotrypsin isoforms (Kh1, Kh2, and Kh3), and two elastase isoforms (E1 and E2) were isolated from the pancreas of European catfish Silurus glanis L. by salting out with (NH4)2SO4, gel chromatography on Sephadex G-75, and ion exchange chromatography on DEAE cellulose. Isoelectric points of the enzymes, determined by isoelectric focusing, amounted to 4.42 for T1, 5.64 for T2, 6.90 for T3, 4.93 for Khl, 5.23 for Kh2, 6.18 for Kh3, 6.17 for E1, and 8.48 for E2. Molecular weights of proteinases within each group were close and amounted to 30100 Da for trypsins, 39800 Da for chymotrypsins, and 24000 Da for elastases. The enzymes isolated displayed maximal activities at alkaline pH values. Inhibitor analysis demonstrated that all the proteinases isolated from European catfish pancreas belonged to the serine type.
Subject(s)
Pancreas/enzymology , Serine Endopeptidases/isolation & purification , Animals , Catfishes , Chromatography, Gel , Isoelectric Focusing , Serine Endopeptidases/chemistry , Serine Endopeptidases/metabolismABSTRACT
Three proteinases named as P1, P2 and P3, were isolated from European sheatfish (Silurus glanis L.) gastric mucosa by salting-out of (NH4)2SO4, gel-chromatography on Sephadex G-75 and ion-exchange chromatography on DEAE-cellulose. Isoelectric points of isolated proteinases were determined by isoelectric focusing and were equal to 1.9, 3.2 and 4.75 respectively for P1, P2 and P3. The molecular weight of P1 was 39,800 Da and proteinases P2 and P3 had a molecular weight of 30,200 Da. The optimum pH for three peptidases isolated from sheatfish gastric mucosa and maximum stability of these enzymes were found at acidic pH. It allowed identifying these proteinases as pepsin-type enzymes of fish.
Subject(s)
Endopeptidases/metabolism , Gastric Mucosa/enzymology , Animals , Chromatography, Gel , Endopeptidases/chemistry , Endopeptidases/isolation & purification , Enzyme Stability , Fishes , Hydrogen-Ion Concentration , Molecular WeightABSTRACT
Surgical and narcotic aggression lead to certain injuries of hepatocytes. The authors followed up the time course of liver injury marker enzymes in two groups of patients subjected to open cholecystectomy (n = 62): succinate dehydrogenase (SDH), glutamate dehydrogenase (GIDH), SGPT, and SGOT. In group 1 the operative trauma was minimal, in group 2 traumatism was higher because of technological difficulties. A high diagnostic significance of SDH and GIDH is worthy of note: their activities increase in proportion with the severity of surgical trauma. Comparison of laboratory data helps objectively assess the severity of operative injury in open cholecystectomy.
