Properties of Extracellular Proteinase of the Micromycete Aspergillus ustus 1 and Its High Activity during Fibrillary-Proteins Hydrolysis

Popova, E. A.; Kreyer, V. G.; Komarevtsev, S. K.; Shabunin, S. V.; Osmolovskiy, A. A.

doi:10.1134/s0003683821020125

Cited by 11 publications

(15 citation statements)

References 15 publications

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“…Microorganisms exhibit a significant role in the mass-production of highly specific, low-cost fibrinolytic enzymes with feasibility of genetic modification through biotechnological approaches. During the past decades, numerous such fibrinolytic enzymes have been tested, specifically from genera Bacillus [2,5,14,15,18,[25][26][27][28][29] and Aspergillus [8,19,20,[30][31][32][33][34]. In addition, fibrinolytic enzymes with varied biochemical characteristics were obtained from bacterial species such as Streptococcus hemolyticus (Streptokinase, exudates of infected wounds) [35], Bacillus subtilis (Nattokinase, Fermented soybeans) [12], Staphylococcus aureus (Staphylokinase, human skin) [36], Bacillus sp.…”

Section: Microbial Fibrinolytic Enzymes: Production Status and Diversitymentioning

confidence: 99%

“…Another method for in-laboratory purification of fibrinolytic enzymes is isoelectrofocusing. The method was used for obtaining pure enzymes for their characterisation and further special studies [20,30,32,64,99]. Isoelectrofocusing allows one not only to isolate enzymes, but also to get a primary idea of their properties and compare fibrinolytic enzymes from different producers.…”

Section: Recovery and Purification Of Fibrinolytic Enzymesmentioning

confidence: 99%

“…strain CK 11-4, respectively, was found to be pH 10.5 [42,43]. Fungal enzymes, such as Aspergillus ustus 1, Arthrobotrys longa 1 and some others are able to hydrolyse fibrin at pH 6.0 [30,176]. Optimal temperature widely ranged from 20-70 • C [20,31,42,100,101,177,180], mostly approx.…”

Section: Physicochemical Characterisation Of Microbial Fibrinolytic Enzymesmentioning

confidence: 99%

“…Optimal temperature widely ranged from 20-70 • C [20,31,42,100,101,177,180], mostly approx. 50 • C for bacterial proteases [5,181] and 37-45 • C for some fungal species [20,30,62,101,104,107]. The molecular weight of fibrinolytic enzymes of different origins broadly varied from 14 kDa to 97 kDa [56,140,172], mostly within 27 kDa and 44 kDa [5,9,182,183].…”

Section: Physicochemical Characterisation Of Microbial Fibrinolytic Enzymesmentioning

confidence: 99%

“…Some fibrinolytic metalloproteases require divalent ions Co 2+ , Ni 2+ , Zn 2+ for their activity [42,185]. Among inhibitors, Di-isopropyl fluorophosphate (DFP) and phenyl methyl sulfonyl fluoride (PMSF) are the frequently used irreversible serine protease inhibitors [5,19,20,27,[30][31][32]63,92]. Inhibitors such as ethylene diamine tetra acetic acid (EDTA), ethyleneglycol bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) and 1, 10 phenanthroline strongly inhibit the fibrinolytic activity of metalloproteases [73,163,183].…”

Section: Physicochemical Characterisation Of Microbial Fibrinolytic Enzymesmentioning

confidence: 99%

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

2021

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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. Fibrinolytic enzymes such as plasminogen activators and plasmin-like proteins hydrolyse thrombi with high efficacy with no significant after-effects and can be cost effectively produced on a large scale with a short generation time. However, the hunt for novel fibrinolytic enzymes necessitates complex purification stages, physiochemical and structural-functional attributes, which provide an insight into their mechanism of action. Besides, strain improvement and molecular technologies such as cloning, overexpression and the construction of genetically modified strains for the enhanced production of fibrinolytic enzymes significantly improve their thrombolytic potential. In addition, the unconventional applicability of some fibrinolytic enzymes paves their way for protein hydrolysis in addition to fibrin/thrombi, blood pressure regulation, anti-microbials, detergent additives for blood stain removal, preventing dental caries, anti-inflammatory and mucolytic expectorant agents. Therefore, this review article encompasses the production, biochemical/structure-function properties, thrombolytic potential and other surplus applications of microbial fibrinolytic enzymes.

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Section: Microbial Fibrinolytic Enzymes: Production Status and Diversitymentioning

confidence: 99%

Section: Recovery and Purification Of Fibrinolytic Enzymesmentioning

confidence: 99%

Section: Physicochemical Characterisation Of Microbial Fibrinolytic Enzymesmentioning

confidence: 99%

Section: Physicochemical Characterisation Of Microbial Fibrinolytic Enzymesmentioning

confidence: 99%

Section: Physicochemical Characterisation Of Microbial Fibrinolytic Enzymesmentioning

confidence: 99%

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

2021

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Thrombolytic and anticoagulant efficiencies of purified fibrinolytic enzyme produced from Cochliobolus hawaiiensis under solid‐state fermentation

Abu‐Tahon,

Abdel‐Majeed,

Ghareib

et al. 2023

Biotech and App Biochem

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Cochliobolus hawaiiensis Alcorn Assiut University Mycological Centre 8606 was chosen from the screened 20 fungal species as the potent producer of fibrinolytic enzyme on skimmed‐milk agar plates. The greatest enzyme yield was attained when the submerged fermentation (SmF) conditions were optimized, and it was around (39.7 U/mg protein). Moreover, upon optimization of fibrinolytic enzyme production under solid‐state fermentation (SSF), the maximum productivity of fibrinolytic enzyme was greatly increased recorded a bout (405 U/mg protein) on sugarcane bagasse, incubation period of 5 days, moisture level of 100%, initial pH of salt basal medium 7.8, incubation temperature at 35°C, and supplementation of the salt basal medium with corn steep liquor (80％, v/v). The yield of fibrinolytic enzyme by C. hawaiiensis under SSF was higher than that of SmF with about 10.20‐fold. The purification procedures of fibrinolytic enzyme by ammonium sulfate (70%), gel filtration, and ion‐exchange columns chromatography caused a great increase in its specific activity to 2581.6 U/mg protein with an overall yield of 55.89%, 6.37 purification fold and molecular weight of 35 kDa. Maximal activity was recorded at pH 7 and 37°C. Significant pH stability was recorded at pH 6.6–7.2, and thermal stability was recorded at 33–41°C. The enzyme showed the highest affinity toward fibrin, with Vmax of 240 U/mL and an apparent Km value of 47.61 mmol. Mg2+ and Ca2+ moderately induced fibrinolytic activity, whereas Cu2+ and Zn2+ greatly suppressed the enzyme activity. The produced enzyme is categorized as serine protease and non‐metalloprotease. The purified fibrinolytic enzyme showed efficient thrombolytic and antiplatelet aggregation activities by completely prevention and dissolution of the blood clot which confirmed by microscopic examination and amelioration of blood coagulation assays. These findings suggested that the produced fibrinolytic enzyme is a promising agent in management of blood coagulation disorders.

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Production and Properties of Aspergillus fumigatus Collagenolytic Proteinase

Nasibov,

Nikitina

2023

Pharm Chem J

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Properties of Extracellular Proteinase of the Micromycete Aspergillus ustus 1 and Its High Activity during Fibrillary-Proteins Hydrolysis

Cited by 11 publications

References 15 publications

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

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

Thrombolytic and anticoagulant efficiencies of purified fibrinolytic enzyme produced from Cochliobolus hawaiiensis under solid‐state fermentation

Production and Properties of Aspergillus fumigatus Collagenolytic Proteinase

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