[Morphological and Physiological Properties of the Micromycete Arthrobotrys longa, a Producer of Longolytin, a Proteolytic Complex with a Thrombolytic Effect].

Production of proteinases with plasmin-like and plasminogen-activating activities by a micromycete Arthrobotrys longa 1 was studied. Polycyclic growth of the producer in submerged cultures was observed, with an endogenous rhythm of the periods of intense microconidia formation alternating with the phases of drastic increase in the content of producing mycelium. The highest plasminogen-like and plasminogen-activating activities (up to 1000 and 500 cond. U/mL, respectively) were found to correlate with the polycyclic growth of the producer, coinciding with the stage of microconidia germination. Enhanced secretion of proteinases with plasminogen-like and plasminogen-activating activity was found to be associated with increased specific growth rate of A. longa l.

[Screening of Producers of Proteinases with Fibrinolytic and Collagenolytic Activities among Micromycetes].

Screening for producers of proteinases with fibrinolytic (plasmin-like and plasminogen-activating) and collagenolytic activities-was carried out among 83 strains of microscopic fungi belonging to various ecological groups. Entomopathogenic micromycetes secreted proteinases with higher fibrinolytic and collagenolytic activity than saprotrophic, potentially phytopathogenic, and epiphytic strains. Micromycete strains possessing proteolytic enzymes with collagenase activity were revealed, as well as the strains producing proteinases with plasmin-like activity. None of the strains studied secreted proteinases possessing only plasminogen-activating activity. Tolypocladium inflatum k1 was found to be a producer of extracellular proteinases with high plasminogen-activating, plasmin-like, and collagenolytic activities. The specific plasminogen-activating activity of T. inflatum k1 was shown to be 20% higher than its plasmin-like activity.

Experimental thrombosis in rabbit marginal ear vein and evaluation of the thrombolytic action of longolytin.

A model of venous thrombosis on the rabbit marginal ear vein was developed. It is based on occlusion of a segment of the left or right ear vein followed by injection of thrombin into this segment. Thrombolytic activity of longolytin applied externally onto the thrombotic venous segment was evaluated. The drug 2-fold shortened the time of clot lysis and 4-4.5-fold increased the rate of thrombolysis. The effect of longolytin was potentiated by heparin. The rate of lysis was higher in large thrombi and in the left vein (effect of asymmetry).

[L-Glutamate oxidase from Streptomyces cremeus 510 MGU: effect of nitrogen sources on enzyme secretion]. / L-Glutamatoksidaza iz Streptomyces cremeus 510 MGU: vliianie istochnikov azota na obrazovanie fermenta.

Effect of nitrogen sources (organic complexes and mineral salts) on L-glutamate oxidase synthesis by Streptomyces cremeus 510 MGU was studied. Optimal enzyme production was not provided by any single nitrogen source. The most effective combination of nitrogen sources (soy flour, peptone, ammonium sulfate) was elaborated by the mathematical planning method. The results of experiment allowed to enhance biosynthesis of extracellular L-glutamate oxidase to 2.4-2.6 U/mL. It was shown that L-glutamate oxidase of Streptomyces cremeus 510 MGU is highly specific to substance and is stable during storage of filtrated culture with pH 6.8-9.0.

[L-Glutamate oxidase from Streptomyces cremeus 510 MGU: growth media optimization for the enhancement of the producer fermentative activity]. / L-glutamatoksidaza iz Streptomyces cremeus 510 MGU: povyshenie fermentativnoi aktivnosti produtsenta putem optimizatsii fermentatsionnoi sredy.

The investigation was devoted to culture conditions optimization aimed to maximum secretion of extracellular L-glutamate oxidase by Streptomyces cremeus 510 MGU. It was shown that Ca ions at the concentration 5-20 mM and 0.1% ammonium sulphate enhanced activity of extracellular enzyme to 4 folds. L-glutamate acid supplement had no effect on enzyme activity. Influence of some bivalent cations and aeration regimes on L-glutamate oxidase activity was investigated. Growth media optimization along with screening of active variants resulted with isolation of the strain with L-glutamate oxidase activity about 2 U/mL Rate of peroxide degradation in the presence of filtrated culture of S. cremeus was determined by chemiluminescence method.

[Physico-chemical properties of the thrombolytic compound longolytin]. / Fiziko-khimicheskie svoistva tromboliticheskogo preparata longolitina.

A preparation exhibiting high fibrinolytic activity and ability to activate plasminogen was isolated from cultivation medium of Arthrobotrys longa. Homogeneous protein, obtained after gel filtration on Sephadex G-100, had molecular mass 28,600, pI-3.68-3.74, optimum activity at pH 6.0-9.0 and temperature optimum at 37 degrees. The enzyme proved to be serine proteinase as shown by analysis using inhibitors; it required thiol groups.

[Effect of the enzyme preparation longolytin on fibrinolysis in animals]. / Vliianie fermentnogo preparata longolitina na fibrinoliz u zhivotnykh.

It has been established that fibrinolytically active enzyme longolytine isolated from the culture fluid of the saprophyte fungus Arthrobotrys longa at intravenous injection favours the prolonged increase of the plasma fibrinolytic properties as well as activation of endogenic plasminogen. Maximum values of fibrinolytic activity have been marked in 5 and 30 min after enzyme intravenous injection. The plasminogen activity is high in 120 min. The fibrinolysis indexes--fibrinolytic activity of the euglobulin fraction and amount of plasminogen activator--in 3,5 and 5 times higher than in vitro at intravenous injection. The activation of coagulation system does not occur.