Targeting mono- and dual-species biofilms of Staphylococcus aureus and Pseudomonas aeruginosa by the recombinant anticoagulant enzyme PAPC from micromycete Aspergillus ochraceus.

Microbial biofilms have recently emerged as a critical target for treating bacterial infections due to their crucial role in developing antibiotic resistance. The wide-spectrum activity of proteolytic enzymes makes them particularly suitable for disrupting biofilms formed by diverse bacterial species, including dual-species biofilms. In this study, we propose the Protease-Activator of Protein C (PAPC) of human blood plasma, an enzyme produced by the micromycete Aspergillus ochraceus, as a novel tool to degrade the protein scaffold of mono- or dual-species biofilms formed by Staphylococcus aureus and Pseudomonas aeruginosa.

Draft genome sequence of VKM-F4104D (L-1) producing peptidase activator of protein C.

VKM-F4104D (L-1) is a saprotrophic fungus isolated from buried soils of Phanagoria (Russia). This strain is known as a producer of the fibrinolytic peptidase-activating plasma protein C. We have sequenced and assembled its genome for a more detailed understanding of the fungus' physiology and encoded peptidases.

Fibrin and Fibrinolytic Enzyme Cascade in Thrombosis: Unravelling the Role.

Blood clot formation in blood vessels (thrombosis) is a major cause of life-threatening cardiovascular diseases. These clots are formed by αA-, βB-, and ϒ-peptide chains of fibrinogen joined together by isopeptide bonds with the help of blood coagulation factor XIIIa. These clot structures are altered by various factors such as thrombin, platelets, transglutaminase, DNA, histones, and red blood cells.

Hemostatically Active Proteinase Produced by : Key Specific Properties and Effect on Target Proteins.

The effect of proteinase on the proteins of the human hemostasis system, fibrin, fibrinogen, plasminogen, protein C, and factor X, was studied. These proteins are key targets for proteolytic enzymes in therapy and diagnosis of thromboembolic complications. It was shown that proteinase efficiently cleaves fibrin and fibrinogen, but does not act precisely, since it cuts all three subunits of these proteins.

Patented Keratinolytic Enzymes for Industrial Application: An Overview.

Proteases that perform keratin hydrolysis (keratinases) have great potential in biotechnology. After investigation, the next step to an industrial application is protecting intellectual property by patenting. There are many fields of discovered keratinase implementation dictated by features of the molecule and its producer.

Keratinolytic Properties of Promising for Biodegradation.

The high demand for keratinolytic enzymes and the modest presentation of fungal keratinase diversity studies in scientific sources cause a significant interest in identifying new fungal strains of keratinase producers, isolating new enzymes and studying their properties. Four out of the 32 cultures showed a promising target activity on protein-containing agar plates- A6, VKPM F-1593, 247, and 1779. The highest values of keratinolytic activity were demonstrated by extracellular proteins synthesized by VKPM F-1593 cultivated under submerged conditions on a medium containing milled chicken feathers.

Chicken Feather Waste Valorization Into Nutritive Protein Hydrolysate: Role of Novel Thermostable Keratinase From RSA27.

Microbial keratinases exhibit a momentous role in converting keratin biowastes into exceedingly valuable protein supplements. This study reports a novel, highly stable keratinase from RSA27 for the production of pure peptides rich in essential amino acids from chicken feathers. Purified keratinase showed a specific activity of 38.

Sustainable Applications of Animal Waste Proteins.

Currently, the growth of the global population leads to an increase in demand for agricultural products. Expanding the obtaining and consumption of food products results in a scale up in the amount of by-products formed, the development of processing methods for which is becoming an urgent task of modern science. Collagen and keratin make up a significant part of the animal origin protein waste, and the potential for their biotechnological application is almost inexhaustible.

Microbial Fibrinolytic Enzymes as Anti-Thrombotics: Production, Characterisation and Prodigious Biopharmaceutical Applications.

Cardiac disorders such as acute myocardial infarction, embolism and stroke are primarily attributed to excessive fibrin accumulation in the blood vessels, usually consequential in thrombosis. Numerous methodologies including the use of anti-coagulants, anti-platelet drugs, surgical operations and fibrinolytic enzymes are employed for the dissolution of fibrin clots and hence ameliorate thrombosis. Microbial fibrinolytic enzymes have attracted much more attention in the management of cardiovascular disorders than typical anti-thrombotic strategies because of the undesirable after-effects and high expense of the latter.

Acute Toxicity, Immunotoxicity and Allergenicity of Protease Complex Obtained from Micromycete .

The different effects on animals of the thrombolytic protease complex of the new producer 203 were studied. The tests of acute toxicity, immunotoxicity and allergenicity should conclude that the studied proteolytic complex is safe for medical usage. For the intravenous and the intraperitoneal routes of administration, the maximum tolerated dose and the median lethal dose were not determined.

Gene Analysis, Cloning, and Heterologous Expression of Protease from a Micromycete Capable of Activating Protein C of Blood Plasma.

Micromycetes are known to secrete numerous enzymes of biotechnological and medical potential. Fibrinolytic protease-activator of protein C (PAPC) of blood plasma from micromycete VKM-F4104D was obtained in recombinant form utilising the bacterial expression system. This enzyme, which belongs to the proteinase-K-like proteases, is similar to the proteases encoded in the genomes of ATCC MYA-4609, ATCC 42149 and 28.

Action of Extracellular Proteases of and Micromycetes on Plasma Hemostasis Proteins.

In this study, we investigated the properties of proteolytic enzymes of two species of , 1 (with a high degree of pathogenicity) and L-1 (a conditional pathogen), and their effects on various components of the hemostasis system (in vitro) in the case of their penetration into the bloodstream. We showed that micromycete proteases were highly active in cleaving both globular (albuminolysis) and fibrillar (fibrin) proteins, and, to varying degrees, they could coagulate the plasma of humans and animals (due to proteolysis of factors of the blood coagulation cascade) but were not able to coagulate fibrinogen. The proteases of both fully hydrolyzed thrombi in 120-180 min.

Vermiculite as a new carrier for extracellular protease production by spp. under solid-state fermentation.

A new method has been developed to increase the productivity of aspergilli - producers of extracellular proteinases based on their cultivation on vermiculite under solid-state fermentation conditions. The productivity of the mycelium L-1 and 1 was 3-18 times higher not only in comparison with submerged cultivation, but also in comparison with growth on other carriers studied under solid-state fermentation conditions. Vermiculite can be considered as a new promising carrier for solid-state fermentation of micromycetes.

Combined microbiological approach to screening of producers of proteases with hemostasis system proteins activity among micromycetes.

A scheme for screening of micromycetes - producers of proteases with the activity of hemostasis system proteins, based on their enzymatic indices determination and the activity towards chromogenic peptide substrates for proteins of the hemostasis system was developed. Depending on the ability of proteases producers to cleave such substrates, an enzymatic reaction in conditions containing human plasma is suggested, which makes it possible to identify the potentiality of the target plasma hemostasis proenzymes activation.