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

Osmolovskiy, A. A.; Lukianova, Anna A.; Звонарева, Е.С.; Kreyer, V. G.; Баранова, Н. А.; Егоров, Н.С.

doi:10.1016/j.btre.2018.e00265

Cited by 4 publications

(6 citation statements)

References 12 publications

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“…Extracellular metabolites play an important role in the vital activity of micromycetes, participating in attachment to the surface and nutrition. Proteolytic activity has already been defined by our investigation group for some of the selected Aspergillus strains [1,2,6,12], but this study provides more systematic and complete information about proteolytic potential of tested Aspergillus isolates. All selected strains except A. phoenicis showed some proteolytic activity.…”

Section: Discussionmentioning

confidence: 99%

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A Novel Approach for Assessing the Proteolytic Potential of Filamentous Fungi on the Example of Aspergillus spp.

Shestakova,

Osmolovskiy,

Lavrenova

et al. 2023

Microbiol. Biotechnol. Lett.

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Proteolytic enzymes produced by filamentous fungi can degrade various fibrous and globular proteins along with other metabolites that may also find application in biotechnology. In this study, the effect of proteolytic enzymes of 22 Aspergillus strains on various proteins was investigated using protein-containing diagnostic media. Subsequently, a new parameter estimating secreted proteinases specificity towards fibrous or globular proteins without its advanced biochemical research -index of severity of proteolytic action (ISPA) -was suggested. This index determines mycozymes specificity in following manner: its value increases with greater affinity to fibrous proteins, decreases if there is higher affinity to globular proteins. ISPA value was the lowest (0.52) for Aspergillus domesticus, indicating the highest specificity to globular proteins, the highest one (1.26) for A. glaucus, whose proteinases best hydrolyzed fibrous proteins. However, the highest overall proteolytic potential was observed for Aspergillus melleus. The ability to produce acid, alkali and extracellular pigments was evaluated for all isolated strains as well.

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Section: Discussionmentioning

confidence: 99%

“…For protease production assessment, a previously developed technique with slight modifications was used [2]. Briefly, a strain was plated on the diagnostic media, containing different protein components indicated in Table 2.…”

Section: Screening For Protease Productionmentioning

confidence: 99%

A Novel Approach for Assessing the Proteolytic Potential of Filamentous Fungi on the Example of Aspergillus spp.

Shestakova,

Osmolovskiy,

Lavrenova

et al. 2023

Microbiol. Biotechnol. Lett.

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“…Then, part of the biomass was transferred to the fermentation medium and cultured for another 2 days. The cultivation was carried out in 750 mL shake flasks containing 100 mL of culture medium [ 19 ]. The preparation of extracellular micromycete proteases were obtained by precipitation of proteins of the filtrate of the culture liquid with ammonium sulfate at the rate of 608 g (NH 4 ) 2 SO 4 per 1 L of sample.…”

Section: Methodsmentioning

confidence: 99%

Action of Extracellular Proteases of Aspergillus flavus and Aspergillus ochraceus Micromycetes on Plasma Hemostasis Proteins

Osmolovskiy

Schmidt

Orekhova

et al. 2021

Life

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In this study, we investigated the properties of proteolytic enzymes of two species of Aspergillus, Aspergillus flavus 1 (with a high degree of pathogenicity) and Aspergillus ochraceus 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 Aspergillus fully hydrolyzed thrombi in 120–180 min. Micromycetes did not show hemolytic activity but were able to break down hemoglobin.

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“…This property of extracellular proteases of micromycetes to activate that factor in human blood plasma has been shown for the first time [17]. This ability distinguishes A. terreus 2 protease from A. ochraceus L-1 protease, which can activate protein C and factor X and finds the application in the creation of diagnostic kits for determining the content of these proteins in patients plasma [7,11,12].…”

Section: Hemostatic Inhibitors Effectmentioning

confidence: 99%

“…Despite the variety of their properties, such as substrate specificity, optimal pH and temperature parameters, some proteases of Aspergillus are able to substantially hydrolyze fibrin or coagulate blood plasma, exhibiting a hemostatic-active effect [5−10]. Proteolytic enzymes of Aspergillus are known for their definite fibrinolytic activity, as well as for the activating effect of such important enzymes of the human hemostasis system as protein C and factor X [5,11,12]. Alkaline exoproteases of Aspergillus terreus have found their practical application in many areas of industry and economy [13−15].…”

Section: Introductionmentioning

confidence: 99%

Action of Extracellular Protease of Aspergillus terreus on Human Plasma Hemostasis Proteins

Osmolovskiy

Звонарева

Баранова

et al. 2022

Microbiol. Biotechnol. Lett.

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Proteolytic enzymes secreted by Aspergillus, as pathogenicity factors, affect blood coagulation and fibrinolysis, and therefore the target proteins of their action in the bloodstream are of significant interest. In the present study, the action of the isolated protease of A. terreus 2 on different human plasma proteins was shown. The protease of A. terreus 2 exhibited the highest proteolytic activity against hemoglobin, which was 2.5 times higher than the albuminolytic activity shown in both of the protein substrates used. In addition, the protease has significant ability to hydrolyze both fibrin and fibrinogen. However, the inability of the A. terreus 2 protease to coagulate rabbit blood plasma and coagulate human and bovine fibrinogen indicates the severity of the enzyme's action on human blood coagulation factors. It should be considered as a potential indicator of this isolated protease's participation in fungal pathogenesis. The protease shows no hemolytic activity. Furthermore, its activity is insignificantly inhibited by thrombin inhibitors, and is not inhibited by plasmin inhibitors.

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

Abstract: HighlightsMost of micromycetes are produced proteolytic enzymes which can affect human hemostasis proteins.For determination of such activity special chromogenic peptide substrates are used.Proteases secretion by micromycetes depends on medium composition.

Cited by 4 publications

References 12 publications

A Novel Approach for Assessing the Proteolytic Potential of Filamentous Fungi on the Example of Aspergillus spp.

A Novel Approach for Assessing the Proteolytic Potential of Filamentous Fungi on the Example of Aspergillus spp.

Action of Extracellular Proteases of Aspergillus flavus and Aspergillus ochraceus Micromycetes on Plasma Hemostasis Proteins

Action of Extracellular Protease of Aspergillus terreus on Human Plasma Hemostasis Proteins

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