Total Proteolytic Activity Research Articles

Effects of Ultrasound on Microstructure and Enzyme Penetration in Beef Longissimus lumborum Muscle

The present study aims to introduce a novel process for improving enzyme diffusion in meat during artificial tenderization. For this purpose, Longissimus lumborum muscles from five young Holstein bulls were cut into 3 × 3 × 3 cm pieces. The samples thus prepared were then treated with 0.1% papain solution and the ultrasonic probe (20 kHz; at 100 and 300 W for 10, 20, and 30 min) under different experimental conditions. For each treatment, total proteolytic activity and muscle microstructure were studied using light and field emission scanning electron microscopy techniques while sensory properties were also investigated. Results indicated that sonication power and duration had significant effects on enzyme activity. The highest activity was observed when an ultrasound radiation of 100 W was applied for 20 min. Compared to the untreated meat, ultrasonically treated meat, and meat immersed in sonicated papain, the deepest part of the sample exposed simultaneously to both enzyme and ultrasound treatments recorded proteolytic activities higher by 62, 25, and 42%, respectively. Although all the treatments caused microstructural damages in beef, the most severe changes and the highest scores for tenderness and flavor were achieved through simultaneous application of papain and ultrasound, which indicates papain penetration into the internal parts of meat. Based on the results obtained, ultrasound technology may be recommended as a useful means to achieve enhanced enzyme diffusion in meat.

Secretion of Proteinases with Fibrinolytic Activity by Micromycetes of the Genus Aspergillus

Proteolytic activity of extracellular enzymes of 11 strains of different Aspergillus species was studied. Comparison of the enzymatic indices of strains grown on agar medium containing either casein or fibrin allowed the selection of the strain Aspergillus terreus 2 as a promising producer of fibrinolytic proteases. It was found that A. terreus 2 proteinases demonstrated maximum activity at pH 8.0. The highest values of fibrinolytic and total proteolytic activities expressed in UTyr (amount of micromoles of tyrosine released from fibrin or casein for 1 min) were 34.0 and 358.3, respectively. Maximum activities were detected when growing the producer on a medium containing only amine nitrogen sources (fish flour hydrolysate and peptone); however, the amount of extracellular protein and the specific fibrinolytic and total proteolytic activities were greater in the medium containing both mineral and amine nitrogen sources (fish flour hydrolysate and sodium nitrate) than in the medium containing only fish flour hydrolysate and peptone as nitrogen sources.

Total proteolytic activity and concentration of alpha-1 antitrypsin in meconium for assessment of the protease/antiprotease balance

BackgroundDuring intrauterine life, various proteolytic enzymes and their main inhibitor, alpha-1 antitrypsin, accumulate naturally in meconium. A protease/antiprotease balance is required to maintain the biological stability of the environment in which the fetus develops. MethodsThe pool of active proteases was determined using the EnzChek Protease Assay Kit. The concentration of alpha-1 antitrypsin in meconium was measured by enzyme-linked immunosorbent assay. Serial portions of meconium (n=80) were collected from healthy full-term neonates (n=19). ResultsMean concentrations of active proteases and alpha-1 antitrypsin were 1.55 [standard deviation (SD) 1.3]mgg−1 (range 0.15–6.17) and 3.72 (SD 1.78)mgg−1 (range 0.76–8.55), respectively, with significant correlation (Rs=0.32, p=0.004). A significant increase in the concentration of active proteases was found between the first and last meconium portions (p<0.05). The proteases in the last meconium portions had a higher reaction velocity and affinity for the substrate than the proteases in the first meconium portions. The active protease:alpha-1 antitrypsin ratio was <0.5 in all first meconium portions, but was higher in the last meconium portions. ConclusionsStrong correlation between the concentrations of active proteases and alpha-1 antitrypsin in meconium may indicate their mutual interaction in the intrauterine environment. Alpha-1 antitrypsin maintains the protease/antiprotease balance during fetal development.

HOMEOSTATIC DISORDERS IN PATIENTS WITH COMBINATION OF HYPERTONIC DISEASE AND CHRONIC PANCREATITIS

Введення. Практично немає жодного захворювання, у розвитку і перебігу якого вегетативна нервова система не відігравала б ролі Це визначає актуальність дослідницьких процесів підтримки гомеостазу при поєднанні різних захворювань.Мета. Метою даного дослідження було вивчити і порівняти характер порушень вегетативної, антиоксидантної, калікреїн-кінінової системи та параметрів ендогенної інтоксикації у хворих з поєднанням гіпертонічної хвороби (ГХ) і хронічного панкреатиту (ХП).Матеріали та методи. У дослідженні взяли участь 40 пацієнтів з ГХ і супутнім ХП в фазі нестійкої ремісії, 15 практично здорових осіб. Середній вік був (47,7 ± 1,2) років. Дослідження вегетативної регуляції проводилось за допомогою визначення стандартних статистичних і спектральних показників варіабельності серцевого ритму (ВСР). Було проведено лабораторне визначення стандартних параметрів антиоксидантної, калікреїн-кінінової системи і тяжкості ендогенної інтоксикації.Результати. Зниження загальної потужності спектра, збільшення частки гуморально-метаболічних впливів на частоту серцевих скорочень, схильність до асимптоматичної вегетативної реактивності, зниження рівня супероксиддисмутази, глутатіону, прекалікреїну, ALPHA 2-макроглобуліну, кінази II, більш високі рівні каталази, середньомолекулярних пептидів, загальної протеолітичної активності калікреїну плазми спостерігались у хворих з поєднанням гіпертонічної хвороби і хронічного панкреатитом.Висновки. У пацієнтів із гіпертонічною хворобою при приєднанні хронічного панкреатиту виявлено ознаки порушення компенсаторних механізмів і підвищення дисбалансу вегетативної системи зі зменшенням пластичності адаптаційних процесів. Поєднання патологій також супроводжувалось проявами ендогенної інтоксикації, порушеннями антиоксидантної, калікреїн-кінінової системи.

THE EFFECT OF EXOGENOUS GLUCOSAMINE HYDROCHLORIDE AND CHONDROITIN SULFATE ON BIOCHEMICAL PARAMETERS OF BLOOD SERUM AND PERIODONTAL TISSUES DURING EXPERIMENTAL INFLAMMATION

The article discusses the evaluation of the effect of exogenous glucosamine hydrochloride and chondroitin sulfate on biochemical parameters of blood serum and periodontal tissues during experimental inflammation. Experimental model of gingivitis were formed at 40 white Wistar rats, were formed: the control and main groups, further investigated the periodontium in 20 intact animals. Research total proteolytic activity in the blood serum of rats with experimental evidence of periodontal inflammation enhancing its performance throughout the experiment as compared with the data obtained in intact animals. In experimental gingivitis with marked intensification of proteolytic activation of lysosomal enzyme acid phosphatase in the serum and in the periodontal tissue homogenates, indicating that damage and destruction of periodontal membrane cells. Installed, that the application of exogenous glucosamine hydrochloride and chondroitin sulfate in experimental inflammation of the periodontal tissues leads to a normalization of the main markers of inflammation, including General proteolytic activity and acid phosphatase activity, demonstrating anti-inflammatory and parodontopathies effect.

Proteolytic activities in cortex of apical parts of Vicia faba ssp. minor seedling roots during kinetin-induced programmed cell death.

Programmed cell death (PCD) is a crucial process in plant development. In this paper, proteolytically related aspects of kinetin-induced PCD in cortex cells of Vicia faba ssp. minor seedlings were examined using morphological, fluorometric, spectrophotometric, and fluorescence microscopic analyses. Cell viability estimation after 46μM kinetin treatment of seedling roots showed that the number of dying cortex cells increased with treatment duration, reaching maximum after 72h. Weight of the apical root segments increased with time and was about 2.5-fold greater after 96h, while the protein content remained unchanged, compared to the control. The total and cysteine-dependent proteolytic activities fluctuated during 1-96-h treatment, which was not accompanied by the changes in the protein amount, indicating that the absolute protein amounts decreased during kinetin-induced PCD. N-ethylmaleimide (NEM), phenylmethylsulfonyl fluoride (PMSF), and Z-Leu-Leu-Nva-H (MG115), the respective cysteine, serine, and proteasome inhibitors, suppressed kinetin-induced PCD. PMSF significantly decreased serine-dependent proteolytic activities without changing the amount of proteins, unlike NEM and MG115. More pronounced effect of PMSF over NEM indicated that in the root apical segments, the most important proteolytic activity during kinetin-induced PCD was that of serine proteases, while that of cysteine proteases may be important for protein degradation in the last phase of the process. Both NEM and PMSF inhibited apoptotic-like structure formation during kinetin-induced PCD. The level of caspase-3-like activity of β1 proteasome subunit increased after kinetin treatment. Addition of proteasome inhibitor MG-115 reduced the number of dying cells, suggesting that proteasomes might play an important role during kinetin-induced PCD.

Penicillium verrucosum as promising candidate for bioremediation of environment contaminated with synthetic detergent at high concentration

The aim of this study was to investigate the effect of synthetic detergent Merix (Henkel, Krusevac, Serbia), and its particular components—ethoxylated oleyl-cetyl alcohol and sodium tripolyphosphate on the growth and metabolic activity of Penicillium verrucosum. During 19 days of fungal cultivation in Czapek-Dox liquid medium supplemented with or without 0.5% pollutants, the following parameters were observed: pH, the total biomass dry weight, the quantity of free and total organic acids and proteolytic activity. The detergent caused a slight stimulatory (2.41%) effect whereas sodium tripolyphosphate and ethoxylated oleyl-cetyl alcohol provided a slight inhibitory action (0.59 and 2.75%, respectively) on the fungal biomass. The pollutants decreased pH values of the media and the quantity of free organic acids. In contrast, they enhanced the quantity of total organic acids. Proteolytic activity remained nearly unchanged (95.8%) in the presence of detergent and reduced to 80.1% in sodium tripolyphosphate-supplemented medium. In contrast, the enzyme activity sharply increased (260.8%) with ethoxylated oleyl-cetyl alcohol. The obtained results indicate the potential of P. verrucosum in bioremediation of environment contaminated with synthetic detergent taken in high concentration.

Litopenaeus vannamei digestive metallo peptidases compensate for anti-nutritional SBTI in feed

The aim of this work was to study mechanisms of homeostasis of the digestive system of the whiteleg shrimp Litopenaeus vannamei when fed anti-nutritional soybean trypsin inhibitor (SBTI). In a previous work, we tested if the shrimp could respond against SBTI in feed with the increase in the digestive proteolytic activity. This time we tested the time of response against the inhibitor and also how the two major digestive trypsin phenotypes change the proteolytic activity. In groups fed SBTI, the total proteolytic activity, trypsin and chymotrypsin decreased during food transit time (2–4h), but increased at 23h postprandial. Three metallo peptidases absent in the control group were identified in groups fed SBTI; individuals with trypsin phenotype CBA increased the activity of two metallo peptidases at 4h of SBTI ingestion, while, individuals with trypsin phenotype CB increased the activity of three metallo peptidases at 23h postprandial. The first evidence of feedback by the digestive system in the whiteleg shrimp fed SBTI was 1h postprandial, with a concomitant increased in metallo peptidases activity at 4h or 23h depending on the trypsin phenotype in each specimen.

Efficacy of nanoceria for periodontal tissues alteration in glutamate-induced obese rats\u2014multidisciplinary considerations for personalized dentistry and prevention

BackgroundNowadays, we face the global epidemic of obesity, that is known to contribute to the development of many diseases, such as the oral cavity pathologies. Dental and oral pathologies are frequently caused by and overlapped with systemic multifactorial diseases such as obesity being its early indicators and risk factors.The aim was to study the influence of nanoceria on periodontal tissues alteration in glutamate (MSG)-induced obese rats.MethodsWe included 52 Wistar rats of both genders and divided into four groups: newborn rats in group 1 (control) received subcutaneously 8 μl/g saline. Group 2 received 3 to 4 mg/g MSG subcutaneously on the second, fourth, sixth, eighth, and tenth day of life; group 3—intragastric administration of nanocrystalline cerium dioxide at a dose of 1 mg/kg volume of 2.9 ml/kg against the background of glutamate-induced obesity; the fourth group of animals was treated with a solution of sodium citrate intragastric volume of 2.9 ml/kg (solvent of nanocrystalline cerium). We determined the total proteolytic activity, the total antitrypsin activity, the content-free fucose and glycosaminoglycanes (GAG), content of TBA-active of products, the content of oxidation-modified proteins (OMB), and catalase activity in the homogenate of soft periodontal tissues of rats.ResultsIntragastric injection of nanoceria prevents activation of proteolytic processes, reducing the catabolism of glycoproteins and proteoglycans of periodontal tissue in MSG-induced obese rats. Injection of nanoceria prevents activation of proteolytic processes, significantly decreases the total proteolytic activity, and inhibits the activation of free radical oxidation in periodontal tissues of rats compared with MSG-induced obesity model without corrections. Further, it significantly increases the total antitrypsin activity in periodontal tissues by 1.7 times, TBA-reagents by 1.7 times, and content of OMB by 1.4 times compared with glutamate-induced obese animals.ConclusionsMSG-induced obesity triggers periodontal tissue alterations in the rat model. Nanoceria contributes to the corrections of pathological changes in periodontal tissues in glutamate-induced obese rats via balancing protein-inhibitory capacity and reducing the depolymerization of fucosylated proteins and proteoglycans and antioxidative activity.

Peptidase compensation in the digestive system of whiteleg shrimp Penaeus vannamei against dietary Kunitz-type soybean trypsin inhibitor

The aim of this study was to assess the regulatory process of digestive peptidases of crustaceans in the presence of soybean trypsin inhibitor (SBTI). This naturally occurring inhibitor in soybean meal was used to inhibit the activity of digestive serine peptidases of the whiteleg shrimp Litopenaeus vannamei. In vitro, SBTI inhibited the total proteolytic activity and chymotrypsin activity by 65%. Trypsin activity was reduced by SBTI from 40% to 15% from 2 to 4 hr of incubation, which is the average time of residence of feed in the shrimp digestive system. During the bioassays, experimental groups were fed with increasing concentration of supplemental SBTI (1 g kg−1 and 2 g kg−1) and digestive gland and faeces of individual specimens were collected daily. At the end of the bioassay, peptidase activity of digestive gland and faeces was shown, revealing differential inhibition after feeding for 5 days. Several serine peptidases were observed in zymograms, showing a compensation effect on the digestive gland through the activation of peptidases from different catalytic type. These results provide evidence that the shrimp digestive gland can overcome the effect of SBTI by two adaptive mechanisms: synthesis of additional peptidases of the serine class and other unidentified peptidases.

Urokinase-type plasminogen activator modulates mammalian circadian clock phase regulation in tissue-type plasminogen activator knockout mice.

Glutamate phase shifts the circadian clock in the mammalian suprachiasmatic nucleus (SCN) by activating NMDA receptors. Tissue-type plasminogen activator (tPA) gates phase shifts by activating plasmin to generate m(ature) BDNF, which binds TrkB receptors allowing clock phase shifts. Here, we investigate phase shifting in tPA knockout (tPA-/- ; B6.129S2-Plattm1Mlg /J) mice, and identify urokinase-type plasminogen activator (uPA) as an additional circadian clock regulator. Behavioral activity rhythms in tPA-/- mice entrain to a light-dark (LD) cycle and phase shift in response to nocturnal light pulses with no apparent loss in sensitivity. When the LD cycle is inverted, tPA-/- mice take significantly longer to entrain than C57BL/6J wild-type (WT) mice. SCN brain slices from tPA-/- mice exhibit entrained neuronal activity rhythms and phase shift in response to nocturnal glutamate with no change in dose-dependency. Pre-treating slices with the tPA/uPA inhibitor, plasminogen activator inhibitor-1 (PAI-1), inhibits glutamate-induced phase delays in tPA-/- slices. Selective inhibition of uPA with UK122 prevents glutamate-induced phase resetting in tPA-/- but not WT SCN slices. tPA expression is higher at night than the day in WT SCN, while uPA expression remains constant in WT and tPA-/- slices. Casein-plasminogen zymography reveals that neither tPA nor uPA total proteolytic activity is under circadian control in WT or tPA-/- SCN. Finally, tPA-/- SCN tissue has lower mBDNF levels than WT tissue, while UK122 does not affect mBDNF levels in either strain. Together, these results suggest that either tPA or uPA can support photic/glutamatergic phase shifts of the SCN circadian clock, possibly acting through distinct mechanisms.

Correction of melanin proteolytic activity upon modeling of alkali burns of the esophagus

During the esophageal burns first and second degree, were observed the increase of total proteolytic activity and of the main proteinase (metalloproteinases and serine). Experimental melanin correction at a dose of 0.1 mg / kg contributed to the normalization of the pathological increase of total proteolytic activity and the activity of the main proteinase (metalloproteinases and serine), which indicates the normalization of the proteolysis system.

Role of cathepsins D in the midgut of Dysdercus peruvianus.

Hemipteran ancestors probably lost their digestive serine peptidases on adapting to a plant sap diet. On returning to protein ingestion, these insects start using cathepsin (lysosomal) peptidases as digestive enzymes, from which the less known is cathepsin D. Nine of the ten cathepsin D transcribing genes found in Dysdercus peruvianus midgut are expressed exclusively in this tissue and only DpCatD10 is also expressed in other tissues. The main action of cathepsins D is in the first (V1) (from three, V1-3) midgut regions, where 40% of the total proteolytic activity was assigned to aspartic peptidases with an optimum pH of 3.5. The most expressed cathepsins D were identified in the midgut luminal contents by proteomics. The data indicate that D. peruvianus have kept a lysosomal gene expressed in all tissues and evolved another set of genes with a digestive function restricted to midgut. Digestive cathepsins D apparently complement the action of digestive cathepsin L and they are arguably responsible for the hydrolysis of cysteine peptidase inhibitors known to be present in the cotton seeds eaten by the insect, before they meet cathepsin L.

Label-Free Differential Proteomics and Quantification of Exoenzymes from Isolates of the Entomopathogenic Fungus Beauveria bassiana.

Beauveria bassiana is an entomopathogenic fungus that grows both in vivo and in vitro. In vivo it can colonize live insect hosts, and tissue digestion occurs by secreted hydrolytic exoenzymes. It can also colonize dead insect tissue provided this is free from competing microorganisms. Depending on whether the host is alive or dead the expression (quality/quantity) of the exoenzymes may vary. We have grown several isolates of B. bassiana in shaking flasks for 120 h at 25 °C in order to evaluate the maximal exoenzyme production using two diet regimes. As sole carbon, nitrogen, and phosphate sources we used 1% shrimp chitin and either 0.5% w/v of dead intact American cockroach (Periplaneta americana) or their isolated cuticles. This is the first report of a differential proteomics of B. bassiana exoenzymes performed by label-free nano-LC MS/MS. Total proteolytic enzyme activity was mainly due to Pr1A or Pr1B depending on the isolate and the diet regime. The most differentially secreted enzymes were: the cuticle-degrading subtilisin Pr1A, GH13 alpha-glycosidase, glucan endo-1,3-beta-glucosidase, subtilisin-like proteinase Spm1, lipase 1, beta-1,3 exoglucanase, and endo-1,3-beta-glucosidase. Among the B. bassiana isolates analyzed, Bb 678 and Bb BG were the most active in Pr1A secretion.

Isolation and Characterization of Gut Bacterial Proteases Involved in Inducing Pathogenicity of Bacillus thuringiensis Toxin in Cotton Bollworm, Helicoverpa armigera.

Bacillus thuringiensis toxin proteins are deployed in transgenic plants for pest management. The present studies were aimed at characterization of gut bacterial proteases involved in activation of inactive Cry1Ac protoxin (pro-Cry1Ac) to active toxin in Helicoverpa armigera. Bacterial strains were isolated from H. armigera midgut and screened for their proteolytic activation toward pro-Cry1Ac. Among 12 gut bacterial isolates seven isolates showed proteolytic activity, and proteases from three isolates (IVS1, IVS2, and IVS3) were found to be involved in the proteolytic conversion of pro-Cry1Ac into active toxin. The proteases from IVS1, IVS2, and IVS3 isolates were purified to 11.90-, 15.50-, and 17.20-fold, respectively. The optimum pH and temperature for gut bacterial protease activity was 8.0 and 40°C. Maximum inhibition of total proteolytic activity was exerted by phenylmethane sulfonyl fluoride followed by EDTA. Fluorescence zymography revealed that proteases from IVS1, IVS2, and IVS3 were chymotrypsin-like and showing protease band at ~15, 65, and 15 kDa, respectively. Active Cry1Ac formed from processing pro-Cry1Ac by gut bacterial proteases exhibited toxicity toward H. armigera. The gut bacterial isolates IVS1, IVS2, and IVS3 showed homology with B. thuringiensis (CP003763.1), Vibrio fischeri (CP000020.2), and Escherichia coli (CP011342.1), respectively. Proteases produced by midgut bacteria are involved in proteolytic processing of B. thuringiensis protoxin and play a major role in inducing pathogenicity of B. thuringiensis toxins in H. armigera.

Metabolic activity of Aspergillus niger and Fusarium lateritium induced by ethoxylated oleyl cetyl alcohol and their bioremediation and biotechnological potential

The effect of ethoxylated oleyl cetyl alcohol (Henkel, Serbia) on the growth and metabolic activity of Aspergillus niger and Fusarium lateritium was in the focus of this paper. The fungi were isolated from wastewater of Lepenica River (Kragujevac, Serbia) at a place where municipal wastewater discharged into the river. The fungi were grown in Czapek-Dox liquid nutrient medium without and with addition of 0.5% pollutant. The physico-chemical and biochemical changes of pH, total biomass dry weight, quantity of free and total organic acids, proteolytic activity and quality of carbohydrates were evaluated from 4-th to 19-th day of fungal growth. The capacity of fungi to decrease concentration of pollutant in medium was determined by cobalt thiocyanate method. The pollutant caused an inhibitory effect on biomass dry weight of A. niger and F. lateritium for 8.50 and 30.61%, respectively. Among tested fungi, A. niger had the better biodegradation capacity (83%) than F. lateritium (65%). Alkaline protease activity of A. niger enhanced in the presence of pollutant for 7.6% whereas the enzyme of F. lateritium retained about 62.2% activity. Overall, the obtained results indicate the potential application of tested fungi in wastewater treatment, detergent industry and biotechnology.

Digestive enzyme activity in the intestine of Nile tilapia (Oreochromis niloticus L.) under pond and cage farming systems.

The effect of different farming systems (cage, pond) upon digestive enzyme activities of Nile tilapia was evaluated. Juvenile Nile tilapia (87.61±1.52g) were simultaneously cultured in pond and cage systems during 90days. Cages used nutritional biphasic plan (35 and 32% crude protein-CP feeds) and ponds used nutritional triphasic plan (35, 32 and 28% CP feeds). Biometric measurements were monthly performed for adjustments in feeding regimes and removal of intestine tissues to evaluate the performance of enzyme activities. Total proteolytic, amylase and lipase activities were not statistically different between the treatments throughout the periods analyzed (31, 63 and 94days of culture). However, trypsin and chymotrypsin activities were higher with 31 and 63days of culture in fish from pond system, suggesting that natural food may have influenced these activities. A positive correlation was observed between the recommended concentration of essential amino acids for Nile tilapia and specific aminopeptidases activity in fish cage system. Substrate-SDS-PAGE revealed 12 active proteolytic bands in both systems. However, integrated density (ID) values were higher in the bands of ponds. Specimens of either cage or pond exhibited five bands of amylolytic activity. Fish from cage and pond systems showed the highest values of ID within 31days of cultivation. In this study, the complexity of digestive functions could be verified for animals maintained under commercial conditions. Some of the assessed enzymes may show adaptations of their activities and/or expression that allow the fish to achieve a more efficient nutrient assimilation.

Characteristics of the effect of cestodes parasitizing the fish intestine on the activity of the host proteinases

The activity and spectrum of proteinases in the intestines of host fishes change upon infestation with cestodes. Serine proteinases are found to make a greater contribution to the total proteolytic activity. The reduction of proteolytic activity is associated with adsorption of the enzymes of the host on the surface of cestodes, and the increase in the activity is caused by the injury of the intestinal mucosa by the attachment apparatuses of cestodes. The inhibition of proteainase activity indicates the possible participation of microbiota enzymes in protein hydrolyses.

Fibrinolytic and collagenolytic activity of extracellular proteinases of the strains of micromycetes Aspergillus ochraceus L-1 and Aspergillus ustus 1

It was shown that extracellular proteinases produced by the strains of micromycetes A. ochraceus L-1 and A. ustus 1 differ by the activity at various pH as well as by the intensity of the effect on fibrillar proteins. It was revealed that the proteinases of A. ochraceus L-1 demonstrated maximum activity during the growth of the producer in the nitrate-free growth medium (the pH of enzyme reaction was 8.0), whereas those of A. ustus 1 showed maximal activity during the growth of the micromycete in the medium containing sodium nitrate (the pH of enzyme reaction was 6.0). Values of specific fibrinolytic and collagenolytic activities of A. ochraceus L-1 were 2.2 and 1.6 times higher than those of A. ustus 1. At the same time, A. ustus 1 showed very low values of total proteolytic (caseinolytic) activity and had a high ratio of fibrinolytic activity to total proteolytic (caseinolytic) activity (6.92). It makes the strain a promising producer of proteinases, which hydrolyze fibrin and collagen.

Detergent-stable proteases from the antarctic scallop Adamussium colbecki

Detergent-stable proteases were found in crude extract of the Antarctic scallop Adamussium colbecki. The results of gelatin zymography indicated the availability of the proteases which able to act in the presence of SDS, whereas plasmin and miniplasmin maintained their activity toward gelatin only after complete removing of detergent. Total proteolytic activity, represented by casein hydrolysis was within the control level after incubation of crude extract with detergents such as Tween 80 and SDS whereas trypsin activity was significantly reduced.