Purification of fibrinolytic enzyme from Bacillus amyloliquefaciens GUTU06 and properties of the enzyme

Wu, Jialin; Lan, Guangqun; He, Na; He, Laping; Li, Cuiqin; Wang, Xiao; Zeng, Xuefeng

doi:10.1016/j.fochx.2023.100896

Cited by 9 publications

(5 citation statements)

References 39 publications

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“…The enzyme exhibited favorable stability within the pH range of 6.6 to 8.2 for 6 h. In this stable pH range, the enzyme activity remained above 80% after 4 h of incubation and still exceeded 71.2% after 6 h. However, following a 24 h incubation period, the residual enzyme activity was observed to be greater than 60.2% (Figure 4D). As already reported, the optimal pH for mushroom fibrinolytic enzymes was within a pH range of 4 to 9, although most of them had an optimal pH between 7 and 7.8 [14,16,[29][30][31][32][33][34][35]. Fibrinolytic enzymes, such as those from Neurospora sitophila [36] and Pleurotus ostreatus [14], showed similar results with an optimal pH value of 7.4.…”

Section: Effects Of Temperature and Ph On The Activity Of Cfesupporting

confidence: 77%

“…Even after a prolonged incubation period of 4 h at these temperatures, the enzyme maintained an activity level exceeding 71.32% (Figure 4B). However, the relative enzyme activity and thermal stability of the enzyme decreased significantly when the temperature exceeded 47 • C. Optimal temperatures for the majority of fibrinolytic enzymes derived from mushrooms typically fall within the temperature range of 20 • C to 60 • C [29][30][31][32][33][34][35]. The optimal temperature of CFE was similar to that of the fibrinolytic enzymes from Bacillus amyloliquefaciens GUTU06 [32] and Coprinopsis atramentaria [33], which exhibited optimal temperatures of 45 • C and 50 • C, respectively.…”

Section: Effects Of Temperature and Ph On The Activity Of Cfementioning

confidence: 88%

“…However, the relative enzyme activity and thermal stability of the enzyme decreased significantly when the temperature exceeded 47 • C. Optimal temperatures for the majority of fibrinolytic enzymes derived from mushrooms typically fall within the temperature range of 20 • C to 60 • C [29][30][31][32][33][34][35]. The optimal temperature of CFE was similar to that of the fibrinolytic enzymes from Bacillus amyloliquefaciens GUTU06 [32] and Coprinopsis atramentaria [33], which exhibited optimal temperatures of 45 • C and 50 • C, respectively. However, it was higher than that of the fibrinolytic enzymes from Hericium erinaceum [27] and Cordyceps militaris [13], which exhibited optimal temperatures of 30 • C and 37 • C, respectively.…”

Section: Effects Of Temperature and Ph On The Activity Of Cfementioning

confidence: 88%

“…The activity of enzymes is also influenced by pH. Changes in pH not only impact the spatial conformation of enzyme molecules but also affect the ionization and affinity between enzyme and substrate molecules [31][32][33].…”

Section: Effects Of Temperature and Ph On The Activity Of Cfementioning

confidence: 99%

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

Wang,

Liu,

Jing

et al. 2024

Foods

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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.

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Section: Effects Of Temperature and Ph On The Activity Of Cfesupporting

confidence: 77%

Section: Effects Of Temperature and Ph On The Activity Of Cfementioning

confidence: 88%

Section: Effects Of Temperature and Ph On The Activity Of Cfementioning

confidence: 88%

Section: Effects Of Temperature and Ph On The Activity Of Cfementioning

confidence: 99%

See 2 more Smart Citations

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

Wang,

Liu,

Jing

et al. 2024

Foods

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“…As shown in Figures 4 and 5, FELP is most active at pH 9.0 and can maintain good stability in an alkaline environment, suggesting that FELP may be an alkaline protease. Moreover, the optimum pH for FELP is higher than the protease from terrestrial organisms [27,38]. It was observed that FELP is stable at 30-60 • C and can still maintain more than 60% activity after incubation for 2 h, exhibiting excellent thermal stability.…”

Section: Discussionmentioning

confidence: 98%

Purification and Properties of a Plasmin-like Marine Protease from Clamworm (Perinereis aibuhitensis)

Jiang,

Zhang,

Zhang

et al. 2024

Marine Drugs

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Marine organisms are a rich source of enzymes that exhibit excellent biological activity and a wide range of applications. However, there has been limited research on the proteases found in marine mudflat organisms. Based on this background, the marine fibrinolytic enzyme FELP, which was isolated and purified from clamworm (Perinereis aibuhitensis), has exhibited excellent fibrinolytic activity. We demonstrated the FELP with a purification of 10.61-fold by precipitation with ammonium sulfate, ion-exchange chromatography, and gel-filtration chromatography. SDS-PAGE, fibrin plate method, and LC–MS/MS indicated that the molecular weight of FELP is 28.9 kDa and identified FELP as a fibrinolytic enzyme-like protease. FELP displayed the maximum fibrinolytic activity at pH 9 (407 ± 16 mm2) and 50 °C (724 ± 27 mm2) and had excellent stability at pH 7–11 (50%) or 30–60 °C (60%), respectively. The three-dimensional structure of some amino acid residues of FELP was predicted with the SWISS-MODEL. The fibrinolytic and fibrinogenolytic assays showed that the enzyme possessed direct fibrinolytic activity and indirect fibrinolysis via the activation of plasminogen; it could preferentially degrade Aα-chains of fibrinogen, followed by Bβ- and γ-chains. Overall, the fibrinolytic enzyme was successfully purified from Perinereis aibuhitensis, a marine Annelida (phylum), with favorable stability that has strong fibrinolysis activity in vitro. Therefore, FELP appears to be a potent fibrinolytic enzyme with an application that deserves further investigation.

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

Wang,

Peng,

et al. 2024

J Food Sci Technol

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Purification of fibrinolytic enzyme from Bacillus amyloliquefaciens GUTU06 and properties of the enzyme

Cited by 9 publications

References 39 publications

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

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

Purification and Properties of a Plasmin-like Marine Protease from Clamworm (Perinereis aibuhitensis)

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

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