Characterization and Production of Thermostable and Acid-stable Extracellular Fibrinolytic Enzymes from Cordyceps militaris

Kim, Seon-Ah; Son, Hong-Joo; Kim, Keun-Ki; Park, Hyun-Chul; Lee, Sang-Mong; Cho, Byung-Wook; Kim, Yonggyun

doi:10.7852/ijie.2011.22.2.83

Cited by 5 publications

(5 citation statements)

References 24 publications

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“…This result agrees with a study on Bacillus lichniformis B4 with an optimum pH for maximum enzyme activity at 7.0 [8]. The fibrinolytic enzyme from Cordyceps militaris shows maximum enzyme activity at pH 7.0-8.0 [17] and Rhyzopus chinensis 12 showed optimum pH 7.6 [20].…”

Section: Discussionsupporting

confidence: 91%

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Production and Partial Characterization of Fibrinolytic Enzyme From a Soil Isolate Aspergillus Carbonarius S-CSR-0007

M.¹,

Nath²,

Smitha³

et al. 2016

Int J Pharm Pharm Sci

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Objective: This work was undertaken with the aim of isolating and screening fungal soil isolates with fibrinolytic activity.Methods: Soil sample near slaughter house was collected and screened for fibrinolytic activity by using fibrin-agar. Enzyme production was optimized under various parameters like pH, temperature, substrate concentration and purified partially by ammonium sulphate precipitation. The stability of the partially purified enzyme was analyzed under the influence of a wide range of pH, temperature, and substrate concentrations.Results: Among the seven isolates screened, Aspergillus carbonarius S-CSR-0007 exhibited largest clear zone and was selected for further studies. Among the various substrates tested casein was found to support the highest caseinolytic activity of 816 U/ml and fibrinolytic activity of 510 U/ml. The culture supernatant of A. carbonarius S-CSR-0007 was fractionated by ammonium sulfate precipitation followed by dialysis, and maximum activity was obtained in the fraction with 80% ammonium sulfate, with an enzyme activity of 1200 U/ml using tyrosine as standard. The partially purified fibrinolytic enzyme showed optimal activity at 45 °C and pH 7.0. The enzyme was stable up to a temperature of 50 °C and pH 8.0, and the optimum substrate concentration was 4%. Conclusion:The crude enzyme showed high blood clot lysis activity, which may be a good candidate in the pharmaceutical industry. However, more studies need to be carried out to establish its clinical use.

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Section: Discussionsupporting

confidence: 91%

“…Cordyceps militaris showed maximum enzyme production at pH 7.0 [17]. The optimum temperature for the enzyme production in this study was at 40 °C which was relatively high compared to previous reports in Cordyceps militaris, Schizophyllum commune BL23, and Serratia marcescens which were 25 °C, 32 °C and 35 °C respectively.…”

Section: Discussioncontrasting

confidence: 51%

Production and Partial Characterization of Fibrinolytic Enzyme From a Soil Isolate Aspergillus Carbonarius S-CSR-0007

M.¹,

Nath²,

Smitha³

et al. 2016

Int J Pharm Pharm Sci

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show abstract

“…At inoculum volumes higher than 7% V/ V, there was a decline in the productivity. Seon Ah Kom et al study, 2011 stated that maximum fibrinolytic enzyme production was obtained at 35-45 0 C [40]. The experimental results in the current study demonstrated comparable outcomes where the productivity of fibrinolytic enzyme from Bacillus cereus ATCC 43 42 was high at a range of temperature 35-43 0 C. Divalent cations such as Ca +2 , Cu +2 and Mn +2 were found to be optimum cofactors for the productivity of FP during the present study.…”

Section: Discussionsupporting

confidence: 74%

Display and Manufacture of Bacterial Thrombolytic Enzymes from Various Soil Environments in Egypt

Kassab

2024

Preprint

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BACKGROUND: Thromboembolic disorders including Myocardial infarction[ MI] and stroke represent the leading causes of death worldwide. Fibrinolytic enzymes extracted from microbial sources showed high efficacy in the dissolution of different blood clots. AIM OF THE STUDY: Screening and the bacterial recombinant DNA manufacture of bacterial fibrinolytic enzymes from different soil environments in Egypt. METHODOLOGY: A total of 50 soil samples were screened for their fibrinolytic enzyme production. The potent fibrinolytic enzyme-producing bacterial isolates were evaluated for their thrombolytic activities using the plasma plate method. The characterization of the enzyme activity and thermostability were carried out; also, the determination of molecular mass using a mass spectrometer and western blot technique was performed. The purification of the fibrinolytic enzymes was done through the affinity chromatography technique. On the other hand, the production of the purified bacterial fibrinolytic enzyme was achieved via bacterial recombinant DNA technology. Formulation of fibrinolytic enzymes in different dosage forms was tried. RESULTS: In the present study, the purified bacterial fibrinolytic enzyme was approximately 83% pure through affinity chromatography and SDS-PAGE assay. The predominating bacterial isolate secreting fibrinolytic enzymes was found to be Bacillus cereus. The fibrinolytic enzyme activities of 50 soil samples were found to be in the range of 0.167-1.853 U/ ml. The fibrinolytic activity of the extracellular enzyme refined from the culture supernatant of Bacillus cereus was relatively stable over the PH range from 7.4-10 for 18 hours and also it showed stability with the highest productivity at a temperature of 43 0C for nearly 60 minutes. The enzyme was called fibrin-proteinase[ FP] in the present study. The FP degraded the fibrin clots via direct thrombolysis. The specific activity and the molecular mass of the FP were estimated to be nearly 40.71 Units/ mg protein and nearly 30 KDa respectively. Protease activity was noticed to be slightly increased in the presence of Fe+3; While it was inhibited in the presence of EDTA. On the other hand, the FP fibrinolytic activity was enhanced greatly after the addition of monovalent cations such as K+1 and Na +1. The enzyme was also active over the temperature range of 20- 65 0C with the highest activity at 55 0C. The optimum temperature and PH of protease production was at 35-43 0C and PH 8.0-10. Conclusion: During the present study, the thermostable fibrinolytic enzyme FP purified from Bacillus cereus in different soil environments in Egypt was a promising fibrinolytic enzyme because it showed a broader efficacious fibrinolytic activity as well as less adverse effects than the current fibrinolytic enzymes globally. It is recommended in the future advanced exploration of the optimization of suitable formulation of bacterial fibrinolytic enzymes to aid in the dissolution of various blood clots and as a consequence overcoming the high rate of mortality globally.

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“…39 At inoculum volumes higher than 7% V/ V, there was a decline in the productivity. Seon Ah Kom et al study, 2011 stated that maximum fibrinolytic enzyme production was obtained at 35-45 0 C. 40 The experimental results in the current study demonstrated comparable outcomes where the productivity of fibrinolytic enzyme from Bacillus cereus ATCC 43 42 was high at a range of temperature 35-43 0 C. Divalent cations such as Ca +2 , Cu +2 and Mn +2 were found to be optimum cofactors for the productivity of FP during the present study. Hg +2 , Zn +2 and Fe +3 were detected to inhibit the production of fibrinolytic enzymes in the current work.…”

Section: Discussionsupporting

confidence: 72%

Screening and production of thrombolytic enzymes from different soil environments in Egypt 1

Kassab

2024

Preprint

View full text Add to dashboard Cite

BACKGROUND: Thromboembolic disorders including Myocardial infarction[ MI] and stroke represent the leading causes of death worldwide. Fibrinolytic enzymes extracted from microbial sources have shown high efficacy in dissolving various types of blood clots. AIM OF THE STUDY: Screening and bacterial recombinant DNA manufacturing of bacterial fibrinolytic enzymes from different soil environments in Egypt. METHODOLOGY: A total of 50 soil samples were screened for fibrinolytic enzyme production. The potent fibrinolytic enzyme-producing bacterial isolates were evaluated for their thrombolytic activities using the plasma plate method. The characterization of the enzyme activity and thermostability was carried out; also, the determination of molecular mass using mass spectrometry and western blot technique was performed. The purification of the fibrinolyric enzymes was done through affinity chromatography. On the other hand the production of the purified bacterial fibrinolytic enzyme was achieved via bacterial recombinant DNA technology. Formulation of fibrinolytic enzyme in different dosage forms were tried. RESULTS: In the present study the purified bacterial fibrinolytic enzyme was approximately 83% pure through affinity chromatography and SDS-PAGE assay. The predominating bacterial isolate secreting fibrinolytic enzymes found to be Bacillus cereus. The fibrinolytic enzyme activities of 50 soil samples were found to be in the range of 0.167-1.853 U/ ml. The fibrinolytic activity of extracellular enzyme refined from the culture supernatant of Bacillus cereus was relatively stable over PH range from 7.4-10 for 18 hours and also it showed a stability with highest productivity at a temperature of 43 ̊C for nearly 60 minutes. The enzyme was called fibrin-proteinase[ FP] in the present study. The FP degraded the fibrin clots via direct thrombolysis. The specific activity and the molecular mass of the FP were estimated to be nearly 40.71 Units/ mg protein and nearly 30 KDa respectively. Protease activity was noticed to be slightly increased in presence of Fe+3; While it was inhibited in presence of EDTA. On the other hand, the FP fibrinolytic activity was enhanced greatly after addition of monovalent cations such as K+1 and Na +1. Enzyme was also active over temperature range 20- 65 ̊̊C with the highest activity was at 55 ̊C. The optimum temperature and pH of protease production was at 35-43 ̊̊C and pH 8.0-10. Conclusion: During the present study, the thermostable fibrinolytic enzyme FP purified from Bacillus cereus in different soil environments in Egypt was a promising fibrinolytic enzyme because it showed a broader efficacious fibrinolytic activity as well as less adverse effects than the current fibrinolytic enzymes globally. It is recommended in the future advanced exploration of the optimization of suitable formulation of bacterial fibrinolytic enzymes to aid in the dissolution of various blood clots and as a consequence overcoming the high rate of mortality globally.

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Characterization and Production of Thermostable and Acid-stable Extracellular Fibrinolytic Enzymes from Cordyceps militaris

Cited by 5 publications

References 24 publications

Production and Partial Characterization of Fibrinolytic Enzyme From a Soil Isolate Aspergillus Carbonarius S-CSR-0007

Production and Partial Characterization of Fibrinolytic Enzyme From a Soil Isolate Aspergillus Carbonarius S-CSR-0007

Display and Manufacture of Bacterial Thrombolytic Enzymes from Various Soil Environments in Egypt

Screening and production of thrombolytic enzymes from different soil environments in Egypt 1

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