Fibrinolytic Enzyme Research Articles

Fibrinolytic enzyme from Arthrospira platensis and its effects on breast cancer cells: Exploring its potential as an innovative therapy.

Fibrinolytic enzymes are promising in treating cardiovascular diseases due to their capacity to dissolve blood clots. The fibrinolytic enzyme from Arthrospira platensis (FEAP) was purified by ion exchange chromatography to investigate its ability to activate plasminogen, as well as its thrombolytic and fibrinogenolytic potential. Subsequently, two different cytotoxic assays (MTT and NR) and hemolysis test were performed to evaluate FEAP's safety. Furthermore, cell migration and the genotoxic and hemolytic potential were also investigated. The purified enzyme showed thrombus degradation of 43% and thrombolytic action directly on fibrin, which can reduce possible side effects, such as hemorrhage. MTT assay was more sensitive to determine the enzyme cytotoxicity, which decreased the viability of breast cancer tumor cells (Sarcoma-180 and MDA-MB-231) and macrophages (J774A.1). In addition, the enzyme also exhibited non-hemolytic, antimetastatic, and non-genotoxic characteristics. These findings are innovative for a fibrinolytic protease and may indicate that it is safe for people undergoing cancer treatment, reducing side effects such as hemorrhage, in addition to inhibiting tumor cells and preventing metastasis, which can help with chemotherapy treatment.

Substrate specificity analysis of semi-purified fibrinolytic protease of Metabacillus sp. CS-2 to support its potential as a wound debridement agent

Fibrinolytic proteases play an important role in the fibrin degradation process, which is crucial in the treatment of chronic wounds as a debridement agent. Fibrinolytics work by breaking down fibrin tissue that forms as part of a blood clot leading to cell necrosis. Metabacillus sp. CS-2 is known to be one of the potential sources of fibrinolytic protease production with high activity. This study aims to examine the activity and specificity of fibrinolytic proteases produced by Metabacillus sp. CS-2 before and after the ultrafiltration process by zymography. In this study, crude protease extraction was carried out from Metabacillus sp. CS-2 followed by ultrafiltration to improve the purity and activity of the enzyme. The obtained ultra-filtrate protease was then characterized for its specificity to 4 protein substrates, i.e. casein, gelatine, fibrin, and collagen. The results showed that the activity of the fibrinolytic protease enzyme from Metabacillus sp. CS-2 was improved by ultrafiltration from 0.342  0.011 to 0.768 0.014 U/mL min-1. The obtained zymogram confirmed that the protease of Metabacillus sp. CS-2 can degrade all of the protein substrates tested. Interestingly, the most specific substrate for Metabacillus sp. CS-2 was fibrin evidenced by intense clear zone smeared from high to low enzyme sizes. In conclusion, ultrafiltration is proven to be effective in increasing the activity of fibrinolytic proteases. The ability to degrade fibrin and collagen substrates likely supports fibrinolytic protease of the strain to function as a debridement agent in wound healing treatment.

Production, Screening and Purification of a Fibrinolytic Enzyme (Actinokinase) from Streptomyces species

Production, Screening and Purification of a Fibrinolytic Enzyme (Actinokinase) from Streptomyces species

Least hemolytic, 12.6 kDa, plasmin-like fibrinolytic protease from marine Penicillium steckii KU1

A novel fibrinolytic enzyme, from the marine fungus Penicillium steckii KU1, was purified to electrophoretic homogeneity. The fibrinolytic protease was purified to 13.56 times with a specific activity of 57.64 U/mg and final yield of 13.93 %. It was found to be a monomeric protein of 12.6 kDa, having optimum activity at 30 °C and pH 8.0. It is a plasmin-like enzyme, showing resemblance to ATP-dependent zinc metalloprotease with isoelectric point (pI) 8.0. Its activity is enhanced by Zn2+, and inhibited by ethylenediaminetetraacetic acid (EDTA), Co2+ and Fe2+. The enzyme interaction with substrate azocasein was endothermic and with inhibitor EDTA exothermic. The Km, Vmax, Kcat and catalytic efficiency of the enzyme for azocasein were determined to be 142.71 μg mL−1, 285.71 μg min−1 mL−1, 6.35 S−1 and 4.45 × 10−2 S−1 μg−1 mL respectively. It hydrolyzed all three chains of fibrinogen within 9 h, and dissolved fibrin completely within 24 h. 2 mg/mL enzyme could dissolve blood clot completely within 30 min, with negligible hemolysis (2.60 %). Lowering the immunogenicity by the application of natural or engineered small proteins is a strategy to enhance the safety and efficacy of thrombolytic therapy. Hence, the present 12.6 kDa, plasmin-like fibrinolytic enzyme appears worthy of further investigations towards a thrombolytic therapeutic.

Maggot kinase: A novel and cost-effective fibrinolytic enzyme from maggots

Maggot kinase, a novel fibrinolytic enzyme source, has been isolated from fly maggots and comprehensively characterized. Using CM-52 ion exchange chromatography and affinity chromatography, we obtained a highly purified and active form of the enzyme. In particular, the fibrinolytic activity of maggot kinase, evaluated using the fibrin plate method, was found to be 8.98 ± 0.08 × 105 U/mg, demonstrating significant efficacy. Further structural analysis using mass spectrometry revealed that maggot kinase consists of a primary sequence of 226 amino acid residues with a molecular weight of 22.91 kDa. In vitro thrombolytic assays demonstrated the enzyme's remarkable ability to degrade the essential fibrinogen subunits (α, β, and γ), thereby facilitating clot lysis. Notably, our studies in a mouse model underscored the significant in vivo thrombolytic activity of maggot kinase, demonstrating its potential to inhibit thrombosis. The finding is particularly significant considering the widespread use of fly maggots in agriculture and animal husbandry due to their rapid growth cycle and minimal nutritional requirements. Our research highlights the untapped potential of fly maggots as a source for maggot kinase development for antithrombotic drug and functional food applications.

Response surface methodology to optimize the Bacillus subtilis var. sojae semen praeparatum liquid fermentation process for the production of fibrinolytic enzyme

Response surface methodology to optimize the Bacillus subtilis var. sojae semen praeparatum liquid fermentation process for the production of fibrinolytic enzyme

Overproduction Of Fibrinolysis Enzyme Produced By Mutant Pseudomonas Aeruginosa Compared To Wild Type Pseudomonas Aeruginosa

Objective: The aim of the study is to increase the production of fibrinolytic enzyme produced by pseudomonas aeruginosa after mutagenesis with physical mutagens compared to the wild type. Study design: Cross-sectional in descriptive study design and case–control in analytical study design. Backgrounds: P. aeruginosa is a ubiquitous gram-negative aerobe. Its opportunistic pathogen that infects patients with cystic fibrosis, burn, wounds, immunodeficiency, chronic obstructive pulmonary disorder (COPD), cancer. Fibrinolytic enzyme is a factor that lysis fibrin clots. This fibrinolytic factor has prospective use totreat cardiovascular diseases such as stroke and heart attack. Thrombotic disorders are the main cause of death world. Methodology: Study patients, specimens, collection of bacterial isolates. Plasma agar plate assay, Skim milk agar assay, Radial caseinolytic assay, Preparing magnetizing Water. Exposure P. aeruginosa to radioactive sources, lasers and magnetic water. Determination of optimal bacterial isolate for the fibrinolytic enzyme production. Results: Increased when P. aeruginosa were exposed to Na23 at an efficiency of 10 µci. Also exposed to 10 ml of magnetic water during 48 hr., then exposed to a cobalt radioactive source during 3 hr., another radioactive source is Cs137, Co60 and Sr90 in 1 hr. during 48 hours with exposure to laser and magnetized water. Conclusion: Conclude that the overproduction of fibrinolytic enzyme (clot-dissolving enzyme) increased when P. aeruginosa were exposed to Na23, Cs137, Co60 and Sr90 with exposed to laser and magnetized water in certain period.

The Impact of Emotional Responses on Female Reproduction: Fibrinolysis in the Spotlight.

Fibrinolytic enzymes modify various substrates required for tissue remodeling, playing a crucial role in mechanisms underlying resilience, reward processing, ovulation, embryo implantation, and placentation. Individuals with low resilience and reduced reward responsiveness, when exposed to chronic stress, are at increased risk of experiencing a range of negative emotions. Chronic anxiety and melancholia are examples of negative emotions associated with hypercortisolism, while fear and atypical depression are characterized by systemic inflammation. Both cortisol and inflammatory cytokines stimulate the production of plasminogen activator inhibitor-1 (PAI-1), a potent fibrinolysis inhibitor. Chronic anxiety, fear, and depression are among the many hypofibrinolytic conditions increasing the risk of oligo-anovulation, miscarriage, fetal growth restriction, and preeclampsia. Although significant, the impact of negative emotions on implantation is not as obvious as on ovulation or placentation. Other hypofibrinolytic conditions that may affect female reproduction through mechanisms dependent or independent of PAI-1 include metabolic disturbances (e.g., due to consumption of highly palatable foods, often used to alleviate negative affect), inflammation, hyperhomocysteinemia, hypothyroidism, hypercortisolism, antiphospholipid antibodies, and the 4G allele of the PAI-1 gene. Benzodiazepines and antidepressants should be used with caution in the first trimester as this combination may cause malformations. Also, selective serotonin reuptake inhibitors have fibrinolytic properties that increase the risk of bleeding after surgical procedures. Psychological interventions, especially group therapy, are effective in the prevention of reproductive disorders. Controlled trials are needed to test the hypothesis that female reproductive health depends on psychological well-being, a balanced diet and physical activity, suppression of inflammation and autoantibodies, and homocysteine and hormonal homeostasis.

Optimizing DCD Liver Grafts With Prolonged Warm Ischemic Time Using Stabilized Plasmin in a Static Cold Storage Orthotopic Rat Liver Transplant Model.

The clinical success of liver transplantation has led to increased demand, requiring further expansion of the donor pool. Therapeutic interventions to optimize organs from donation after circulatory death (DCD) have significant potential to mitigate the organ shortage. Dysfunction in DCD liver grafts is mediated by microvascular thrombosis during the warm ischemic period, and strategies that reduce this thrombotic burden may improve graft function. We hypothesized that the administration of the fibrinolytic enzyme plasmin to the donor organ during the cold storage period would reduce the thrombotic burden and improve DCD liver graft function. In 2 separate cohorts, 32 syngeneic orthotopic rat liver transplants were performed in Lewis rats. Livers were procured from donors with 45 min of warm ischemic injury. Liver grafts were flushed with histidine-tryptophan-ketoglutarate preservation solution mixed with either plasmin (experimental group) or albumin (control group). All investigators were blinded to treatment group. After preparing the liver for implant using a modified cuff technique, the liver was stored for 1 h by static cold storage at 4 °C. Immediately before implantation, the liver graft was flushed, and this effluent was analyzed for fibrin degradation products to determine graft clot burden. Twenty-four hours following transplantation, animals were euthanized, and samples were collected. Recipient survival was significantly higher for DCD liver grafts treated with plasmin compared with control. Moreover, histology of liver graft tissue immediately before implant reflected significantly reduced congestion in plasmin-treated livers (score, mean ± SD: 0.73 ± 0.59 versus 1.12 ± 0.48; P = 0.0456). The concentration of fibrin degradation products in the final flush before implantation was significantly reduced in plasmin-treated livers (743 ± 136 versus 10 919 ± 4642 pg/mL; P = 0.0001), reflecting decreased clot burden in the graft. The present study demonstrates that plasmin improves survival and may reduce thrombotic burden in DCD liver grafts with prolonged warm ischemic injury, meriting further study.

Computational analysis to comprehend the structure-function properties of fibrinolytic enzymes from Bacillus spp for their efficient integration into industrial applications

BackgroundThe fibrinolytic enzymes from Bacillus sp. are proposed as therapeutics in preventing thrombosis. Computational-based analyses of these enzymes’ amino acid composition, basic physiological properties, presence of functional domain and motifs, and secondary and tertiary structure analyses can lead to developing a specific enzyme with improved catalytic activity and other properties that may increase their therapeutic potential. MethodsThe nucleotide sequences of fibrinolytic enzymes produced by the genus Bacillus and its corresponding protein sequences were retrieved from the NCBI database and aligned using the PRALINE programme. The varied physiochemical parameters and structural and functional analysis of the enzyme sequences were carried out with the ExPASy-ProtParam tool, MEME server, SOPMA, PDBsum tool, CYS-REC tool, SWISS-MODEL, SAVES servers, TMHMM program, GlobPlot, and peptide cutter software. The assessed in-silico data were compared with the published experimental results for validation. ResultsThe alignment of sixty fibrinolytic serine protease enzymes (molecular mass 12–86 kDa) sequences showed 49 enzymes possess a conserved domain with a catalytic triad of Asp196, His242, and Ser569. The predicted instability and aliphatic indexes were 1.94–37.77, and 68.9–93.41, respectively, indicating high thermostability. The random coil means value suggested the predominance of this secondary structure in these proteases. A set of 50 amino acid residues representing motif 3 signifies the Peptidase S8/S53 domain that was invariably observed in 56 sequences. Additionally, 28 sequences have transmembrane helices, with two having the most disordered areas, and they pose 25 enzyme cleavage sites. A comparative analysis of the experimental work with the results of in-silico study put forward the characteristics of the enzyme sequences JF739176.1 and MF677779.1 to be considered when creating a potential mutant enzyme as these sequences are stable at high pH with thermostability and to exhibit αβ-fibrinogenase activity in both experimental and in-silico studies.

Purification and Characterization of a Novel Fibrinolytic Enzyme from Marine Bacterium Bacillus sp. S-3685 Isolated from the South China Sea.

A novel fibrinolytic enzyme, BSFE1, was isolated from the marine bacterium Bacillus sp. S-3685 (GenBank No.: KJ023685) found in the South China Sea. This enzyme, with a molecular weight of approximately 42 kDa and a specific activity of 736.4 U/mg, exhibited its highest activity at 37 °C in a phosphate buffer at pH 8.0. The fibrinolytic enzyme remained stable over a pH range of 7.5 to 10.0 and retained about 76% of its activity after being incubated at 37 °C for 2 h. The Km and Vmax values of the enzyme at 37 °C were determined to be 2.1 μM and 49.0 μmol min-1 mg-1, respectively. The fibrinolytic activity of BSFE1 was enhanced by Na+, Ba2+, K+, Co2+, Mn2+, Al3+, and Cu2+, while it was inhibited by Fe3+, Ca2+, Mg2+, Zn2+, and Fe2+. These findings indicate that the fibrinolytic enzyme isolated in this study exhibits a strong affinity for fibrin. Moreover, the enzyme we have purified demonstrates thrombolytic enzymatic activity. These characteristics make BSFE1 a promising candidate for thrombolytic therapy. In conclusion, the results obtained from this study suggest that our work holds potential in the development of agents for thrombolytic treatment.

Production of Nattokinase from Bacillus amyloliquefaciens MRS18: A Bacterial Strain Isolated from Fermented Beans.

Nattokinase (NK) is a naturally occurring fibrinolytic protease enzyme obtained from the traditional Japanese food called Natto and has several uses in the pharmaceutical and medical industries. Nowadays, the most often used thrombolytic agent in the clinical field is NK, in part because it is less expensive than other thrombolytic medicines. The objective of this study is to investigate the screening, isolation and characterization of the NK enzyme-producing Bacillus strain from fermented Soya beans. The sample of fermented soya beans were tested for the presence of fibrinolytic protease- producing bacteria, followed by the screening, extraction, characterization and clot lysis assays. A total of three isolates were screened for caseinolytic activities by casein hydrolysis assay. Out of those isolates, MRS18 was found to be potent in producing the enzyme proteinase. To determine the taxonomy and phylogeny of these isolates, biochemical and molecular characterization has been carried out. Bacillus amyloliquefaciens MRS18 has been found with the highest caseinolytic activity. The clot lysing ability of the potent strain Bacillus amyloliquefaciens was found to be 61.7% after 120 min, and on further purification, by ammonium sulphate precipitation method, the lysis percentage was found to be 656% after 120 min. From the results of the present study, we concluded that Bacillus amyloliquefaciens isolated from the fermented soya beans produced an NK enzyme that exhibits immense potential to lyse blood clots.

Systematic review: How to obtain Fibrinolytic Enzymes from Fungi

Fibrinolytic complications such as stroke and peripheral occlusive disease due to vascular occlusion are significant causes of poor prognosis and death. Given the ability of mushroom fibrinolytic enzymes to dissolve blood clots and prevent side effects, their use as a potential thrombolytic therapy has attracted great interest. Serine protease, metalloprotease, and serine metalloprotease are protease enzymes that can affect the fibrinolytic mechanism by dissolving blood clots and increasing blood flow. This systematic review aimed to disclose all information regarding the production, purification, and characterization of fibrinolytic proteases from fungi. Searches were conducted on online media databases, including Google and journal sites (NCBI, PubMed, and others, using the keywords "(Fibrinolytic Enzymes) OR (Fibrinolytic Proteases) AND (Fungi or Mushrooms)" from 2012-2023. This systematic review observed reviews from the purification and characterization steps, indicating the need for attention to the manufacturing process and enzymatic applications. The production of these enzymes by fungi is associated with developments in pharmaceutical applications.

Process optimization for the production of fibrinolytic enzyme by a novel marine fungus – Penicillium steckii KU1

The demand for microbial thrombolytic agents is alarmingly increasing, and the discovery of novel microbial species with significant level of fibrinolytic enzyme production are required to cope up with the current circumstances. Surprisingly, marine fungi remain largely unexplored in this regard, presenting an untapped potential for the production of fibrinolytic enzymes.A novel marine fungal strain (Penicillium steckii KU1) was isolated successfully from Kappil beach, Kerala. At the beginning Czapek-dox media was used for the production of fibrinolytic enzyme. Statistical optimization of the enzyme production was achieved initially by Plackett-Burman design (PB). Interactive effect of various process parameters was further scrutinized by Central Composite Design (CCD). Incubation time, inoculum size and concentration of casein were shown significant contribution towards production. Statistical optimization, resulted in a tenfold increment (454.49 ± 14.66 U/mL) [optimized conditions - 30 °C, pH 7, casein 2.7 % (w/v), inoculum 5.75 million spores/mL and 115 h incubation], against the predicted value 441.42 U/mL.

Heterologous expression of nattokinase in E. coli: Biochemical characterization and functional analysis of fibrin binding residues

Heterologous expression of nattokinase, a potent fibrinolytic enzyme, has been successfully carried out in various microorganisms. However, the successful expression of this enzyme as a soluble protein was not achieved in E. coli. This study delves into the expression of nattokinase in E. coli as a soluble protein followed by its biochemical characterization and functional analysis for fibrinolytic activity. E. coli BL21C41 and pET32a vector host strain with pGro7 protein chaperone induced with IPTG at 16 °C 180 rpm for 16 h enabled the production of recombinant nattokinase in soluble fraction. Enzymatic assays demonstrated its protease activity, while characterization revealed optimal catalytic conditions at 37 °C and pH 8.0, with remarkable stability over a broad pH range (6.0–10.0) and up to 50 °C. The kinetic constants were determined as follows: Km = 25.83 ± 3.43 μM, Vmax = 62.91 ± 1.68 μM/s, kcat = 38.45 ± 1.06 s−1, and kcat/Km = 1.49 × 106 M−1 s−1. In addition, the fibrinolytic activity of NK, quantified by the fibrin plate hydrolysis assay was 1038 ± 156 U/ml, with a corresponding specific activity of 1730 ± 260 U/mg and the assessment of clot lysis time on an artificial clot (1 mg) was found to be 51.5 ± 2.5 min unveiling nattokinase's fibrinolytic potential. Through molecular docking, a substantial binding energy of −6.46 kcal/mol was observed between nattokinase and fibrin, indicative of a high binding affinity. Key fibrin binding residues, including Ser300, Leu302, and Asp303, were identified and confirmed. These mutants affected specifically the fibrin binding and not the proteolytic activity of NK. This comprehensive study provides crucial conditions for the expression of protein in soluble form in E. coli and biochemical properties paving the way for future research and potential applications in medicine and biotechnology.

Fibrinolytic enzyme from Arthrospira platensis: Kinetic and thermodynamic investigation

Fibrinolytic enzymes from microorganisms have been investigated by their potential application as thrombolytic agents. Previous studies show that the fibrinolytic enzyme from Arthrospira platensis (FEAP) is stable at human physiological temperature (<50 °C) and pH (6.0–10.0). Understanding of the kinetic/thermodynamic characteristics is important to make the advances for industrial applications feasible. Therefore, a kinetic/thermodynamic analysis of FEAP was performed on the fibrin hydrolysis and enzyme thermoinactivation. Results showed two Michaelis-Menten-type profiles with two plateaus, revealing high affinity for the substrate with low Michaelis constant values (0.02 and 6.37 µg∙mL−1). The activation energy of the hydrolysis catalyzed by FEAP and the standard enthalpy variations of reversible enzyme unfolding were 49.65 and 107.01 kJ∙mol−1, respectively. When the temperature increased from 40 to 70 °C, the deactivation rate constant increased from 0.0050 to 0.0134 min−1, while the half-life decreased from 138.62 to 51.72 min. These results allowed to estimate the activation energy (E*d = 27.50 kJ∙mol−1), enthalpy (24.64 ≤ ΔH*d ≤ 24.89 kJ∙mol−1), entropy (−209.86 ≤ ΔS*d ≤ −210.10 J∙mol−1∙K−1), and Gibbs free energy (90.61 ≤ ΔG*d ≤ 96.74 kJ∙mol−1) of thermal denaturation. The data suggest that FEAP is a promising and thermostable biocatalyst that could be exploited for industrial applications.

Nattokinase attenuates endothelial inflammation through the activation of SRF and THBS1

Natto contains a potent fibrinolytic enzyme called nattokinase (NK), which has thrombolytic, antihypertensive, antiatherosclerotic and lipid-lowering effects. Although NK has been recognized for its beneficial effect on humans with atherosclerotic cardiovascular disease (ASCVD), the underlying mechanisms involved in vascular inflammation-atherosclerosis development remain largely unknown. The current study aimed to explore the effects of NK on gene regulation, autophagy, necroptosis and inflammasome in vascular inflammation. The transcriptional profiles of NK in endothelial cells (ECs) by RNA sequencing (RNA-seq) revealed that NK affected THBS1, SRF and SREBF1 mRNA expression. In Q-PCR analysis, SRF and THBS1 were upregulated but SREBF1 was unaffected in ECs treated with NK. NK treatment induced autophagy and inhibited NLRP3 inflammasome and necroptosis in ECs. Furthermore, the inhibition of SRF or THBS1 by siRNA suppressed autophagy and enhanced the NLRP3 inflammasome and necroptosis. In a mouse model, NK reduced vascular inflammation by activating autophagy and inhibiting NLRP3 inflammasome and necroptosis. Our findings provide the first evidence that NK upregulates SRF and THBS1 genes, subsequently increasing autophagy and decreasing necroptosis and NLRP3 inflammasome formation to reduce vascular inflammation. Therefore, NK could serve as nutraceuticals or adjuvant therapies to reduce vascular inflammation and possible atherosclerosis progression.

Purification and Biochemical Characterization of a Novel Fibrinolytic Enzyme from Culture Supernatant of Coprinus comatus.

A novel fibrinolytic enzyme was produced by the liquid fermentation of Coprinus comatus. The enzyme was purified from the culture supernatant by hydrophobic interactions, gel filtration, and ion exchange chromatographies. It was purified by 241.02-fold, with a specific activity of 3619 U/mg and a final yield of 10.02%. SDS-PAGE analysis confirmed the purity of the enzyme, showing a single band with a molecular weight of 19.5 kDa. The first nine amino acids of the N-terminal of the purified enzyme were A-T-Y-T-G-G-S-Q-T. The enzyme exhibited optimal activity at a temperature of 42 °C and pH 7.6. Its activity was significantly improved by Zn2+, K+, Ca2+, Mn2+, and Mg2+ while being inhibited by Fe2+, Fe3+, Al2+, and Ba2+. The activity of the enzyme was completely inhibited by ethylenediamine tetraacetic acid (EDTA), and it was also dose-dependently inhibited by phenylmethylsulfonyl fluoride (PMSF) and soy trypsin inhibitor (SBTI). However, inhibitors such as N-α-tosyl-L-phenylalanine chloromethyl ketone (TPCK), aprotinin, and pepstatin did not significantly affect its activity, suggesting that the enzyme was a serine-like metalloproteinase. The enzyme acted as both a plasmin-like fibrinolytic enzyme and a plasminogen activator, and it also exhibited the capability to hydrolyze fibrinogen and fibrin. In vitro, it demonstrated the ability to dissolve blood clots and exhibit anticoagulant properties. Furthermore, it was found that the enzyme prolonged activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT), and reduced the levels of fibrinogen (FIB) and prothrombin activity (PA). Based on these studies, the enzyme has great potential to be developed as a natural agent for the prevention and treatment of thrombotic diseases.

Role of gamma radiation as an agent modulator of Mucor subtilissimus UCP1262 Fibrinolytic Enzyme (MsFE)

Thrombosis is a dangerous condition that can result in major damage. Fibrinolytic enzymes are significant therapeutic agents for thrombosis. This study focused on MsFE, a fibrinolytic protease from M. subtilissimus UCP1262, exploring its physicochemical traits and the impact of gamma radiation on its activity. Investigations included: fibrinogenolytic activity, effects of solvents and ions, gamma rays, enzyme shelf life, FTIR, CD, clotted times determination, and enzymatic kinetics. Notably, the major results obtained show the protease retained 90 % activity in organic solvents, FTIR indicated carbonyl functional groups and Circular dichroism spectroscopic studies indicated that the secondary structure (β-sheet) of enzyme remains mostly unaffected over pH 2.0–9.0, whereas considerable changes were observed in the tertiary structure. Kinetic analysis revealed altered parameters post-irradiation. Gamma radiation initially enhanced enzyme activity (10 kGy) but progressively decreased beyond 20 kGy. Metallic ions, especially magnesium, augmented irradiated enzyme activity. This study highlights gamma radiation's potential as a modulator for enhanced fibrinolytic activity, suggesting future biomedical applications.

Valorization of dairy effluent for the conversion of microbial fibrinolytic enzymes for environment management and bio-medical applications

Valorization of dairy effluent for the conversion of microbial fibrinolytic enzymes for environment management and bio-medical applications