Fibrinolytic Bacterial Enzymes with Thrombolytic Activity

Kotb, Essam

doi:10.1007/978-3-642-24980-8

Cited by 24 publications

(3 citation statements)

References 181 publications

(241 reference statements)

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“…They can be easily produced on large scale with short generation time and are less expensive [23][24][25]. Fibrinolytic proteases basically belong to either the metalloproteases or the serine class of protease [26]. Numerous microbial fibrinolytic enzymes have been isolated and characterized from bacterial species, such as Bacillus polymyxa and B. subtilis [27], E. coli [28], Pseudomonas [29], B. cereus IND1 [30], Streptomyces [31], Paenibacillus sp.…”

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Thrombolytic Potential of Novel Thiol-Dependent Fibrinolytic Protease from Bacillus cereus RSA1

et al. 2019

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The present study demonstrates the production and thrombolytic potential of a novel thermostable thiol-dependent fibrinolytic protease by Bacillus cereus RSA1. Statistical optimization of different parameters was accomplished with Plackett-Burman design and validated further by central composite design with 30.75 U/mL protease production. Precipitation and chromatographic approaches resulted in 33.11% recovery with 2.32-fold purification. The molecular weight of fibrinolytic protease was 40 KDa and it exhibited a broad temperature and pH stability range of 20-80 • C and pH 5-10 with utmost activity at 50 • C and pH 8, respectively. The protease retained its fibrinolytic activity in organic solvents and enhanced the activity in solutions with divalent cations (Mn 2+ , Zn 2+ , and Cu 2+ ). The enzyme kinetics revealed K m and V max values of 1.093 mg/mL and 52.39 µg/mL/min, respectively, indicating higher affinity of fibrinolytic activity towards fibrin. Also, complete inhibition of fibrinolytic activity with DFP and a 2-fold increase with DTT and β-mercaptoethanol indicates its thiol-dependent serine protease nature. MALDI-TOF analysis showed 56% amino acid sequence homology with Subtilisin NAT OS = Bacillus subtilis subsp. natto. The fibrinolysis activity was compared with a commercial thrombolytic agent for its therapeutic applicability, and fibrinolytic protease was found highly significant with absolute blood clot dissolution within 4 h in in vitro conditions. The isolated fibrinolytic protease of Bacillus cereus RSA1 is novel and different from other known fibrinolytic proteases with high stability and efficacy, which might have wide medicinal and industrial application as a thrombolytic agent and in blood stain removal, respectively.Biomolecules 2020, 10, 3 2 of 23 (active protease), converting soluble fibrinogen (glycoprotein) into fibrin polymer (insoluble blood clot) [5]. Fibrin clots are hydrolyzed by plasmin [12], which is stimulated from plasminogen by plasminogen activators (PAs) [13,14]. This natural dynamic equilibrium is disturbed when the process of natural fibrin clot hydrolysis undergoes pathophysiological shambles, leading to formation of fibrin clots. Such clots may cause hindrance in the circulation of blood, causing blockage in blood vessels, ultimately leading to cardiac disorders which are life-threatening [15,16].Approaches such as use of anti-coagulant agents, anti-platelet drugs, fibrinolytic enzymes, and surgical operations are employed for the treatment of thrombosis and to dissolve the blood clots [17]. Further, there are numerous side effects which may occur following the administration of the available anti-thrombotic strategies, as well as the high expense, which limit their scope. The effects of reperfusion, urticaria (allergic reaction), and hemorrhage are the major inimical consequences of such thrombolytic approaches on human health [18,19]. Other after effects include headache, dizziness, ulcers, increased clotting time, nausea and vomiting, etc. Management of thrombosis usin...

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Thrombolytic Potential of Novel Thiol-Dependent Fibrinolytic Protease from Bacillus cereus RSA1

et al. 2019

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“…Thrombolytic agents have been identified in numerous organism, ranging from earthworm, snake, centerpede, mushroom, kelp, some algae and microrganisms (Kotb, 2012;Yogesh & Halami, 2017). Hence, streptokinase from Streptococcus hemolyticus and staphylokinase from Staphylococcus aureus which act as plasminogen activator, have been adopted as thrombolytic agent's medicine to overcome thrombosis.…”

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confidence: 99%

“…Hence, streptokinase from Streptococcus hemolyticus and staphylokinase from Staphylococcus aureus which act as plasminogen activator, have been adopted as thrombolytic agent's medicine to overcome thrombosis. Meanwhile, two other agents known to be effective (Kotb, 2012) include tissue-type plasminogen activator (t-PA) (Collen & Lijnen, 2004) and urokinase (Duffy, 2002), which both activate plasminogen into active plasmin, in order to properly degrade fibrin. However, in addition to being expensive, certain side effects have been reported through injection application, which is possibly due to the presence of impurities or some imune active material in the preparation.…”

Section: Resultsmentioning

confidence: 99%