Pseudomonas aeruginosa was isolated from injuries of patients' wounds and burns, and to ensure that the isolate was belonging to P. aeruginosa, several tests were performed, such as staining techniques, a biochemical test, morphological test, Vitek 2 system, and sensitivity test. The results of the gram stain test showed rod pink gram-negative bacteria, demonstrating that the isolate belonged to P. aeruginosa. Growth optimization of bacterial was performed by assessing different combinations of pH and temperatures. It is revealed that the best conditions for increasing the number of bacteria were achieved at 37°C with the bacterial number of 5.53×108 and pH 6 with the bacterial number of 5.87×108. Fibrinolytic enzyme is an agent that lysis fibrin clots. This fibrinolytic factor has prospective use to treat cardiovascular diseases, such as stroke and heart attack. Cardiovascular diseases have attracted worldwide attention for their elevation morbidity and mortality. Fibrinolytic enzyme was extracted by centrifugation at 10000 × g at 4°C for 10 min, the supernatant was kept and the pellet having bacterial cells was discarded. Purification of the fibrinolytic enzyme was achieved using salt precipitation, ion exchange, and gel filtration chromatographic techniques. The results showed that the gel filtration chromatography had optimal specific activity and purification fold at 562.6 U/ml, and the final specific activity of the purified enzyme increased 4.1 times. The molecular weight of the fibrinolytic enzyme was determined at26 kDa by gel filtration chromatography. The purified fibrinolytic enzyme had optimum activity atpH 7 and40°C. The pH stability for the enzyme activity was found in pH 6-7 and the range of 10-40°C. |
- Abdulghafoor B, Mohamed Z, Najla A, Salah A. Ems mutation improve the fibrinolytic enzyme activity in bacillus subtilis strain. World Appl Sci J. 2019;37(2):135-9.
- Ali AMM, Bavisetty SCB. Purification, physicochemical properties, and statistical optimization of fibrinolytic enzymes especially from fermented foods: a comprehensive review. Int J Biol Macromol. 2020.
- Venkata Naga Raju E, Divakar G. An Overview on Microbial Fibrinolytic Proteases. Int J Pharm Life Sci. 2014;5(3).
- Wendelboe AM, Raskob GE. Global burden of thrombosis: epidemiologic aspects. Circ Res. 2016;118(9):1340-7.
- Ali EH. The Effects of Enzyme Nanoparticles on Adhesion of Pathogenic Bacteria. Plant Arch. 2020;20(1):217-9.
- Bédard E, Prévost M, Déziel E. Pseudomonas aeruginosa in premise plumbing of large buildings. Microbiologyopen. 2016;5(6):937-56.
- Sattar RJ, Ali EH. Purification and Characterization of Lipase Production from Pseudomonas Aeruginosaand Study Effect of Silver Nanoparticle in Activity of Enzyme in Application of Biology. Plant Arch. 2019;19(2):4379-85.
- Spilker T, Coenye T, Vandamme P, LiPuma JJ. PCR-based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fibrosis patients. J Clin Microbiol. 2004;42(5):2074-9.
- Keziah SM, Devi CS. Focalization of thrombosis and therapeutic perspectives: a memoir. Orient Pharm Exp Med 2018;18(4):281-98.
- Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72(1-2):248-54.
- Chang C-T, Wang P-M, Hung Y-F, Chung Y-C. Purification and biochemical properties of a fibrinolytic enzyme from Bacillus subtilis-fermented red bean. Food Chem. 2012;133(4):1611-7.
- Afifah DN, Sulchan M, Syah D. Purification and characterization of a fibrinolytic enzyme from Bacillus pumilus 2. g isolated from Gembus, an Indonesian fermented food. Prev Nutr Food Sci. 2014;19(3):213.
- Ali EH, Mohammed KR. Extraction, Purification and Characterization of Peroxidase from Pseudomonas aeruginosa and Utility as Antioxidant and Anticancer. Baghdad Sci J. 2019;16(4).
- Hmood SA, Aziz GM. Optimum conditions for fibrinolytic enzyme (Nattokinase) production by Bacillus sp. B24 using solid state fermentation. Iraqi J Sci. 2016;57(2C):1391-401.
- Chen Y-C, Chiang Y-C, Hsu F-Y, Tsai L-C, Cheng H-L. Structural modeling and further improvement in pH stability and activity of a highly-active xylanase from an uncultured rumen fungus. Bioresour Technol. 2012;123:125-34.
- Bezerra RP, Teixeira JA, Silva FO, Correia JM, Porto TS, Lima-Filho JL, et al. Screening, production and biochemical characterization of a new fibrinolytic enzyme produced by Streptomyces sp.(Streptomycetaceae) isolated from Amazonian lichens. Acta Amazon. 2016;46:323-32.
- Liu X-l, Zheng X-q, Qian P-z, Kopparapu N-k, Deng Y-p, Nonaka M, et al. Purification and characterization of a novel fibrinolytic enzyme from culture supernatant of Pleurotus ostreatus. J Microbiol Biotechnol. 2014;24(2):245-53.
- Cha W-S, Park S-S, Kim S-J, Choi D. Biochemical and enzymatic properties of a fibrinolytic enzyme from Pleurotus eryngii cultivated under solid-state conditions using corn cob. Bioresour Technol. 2010;101(16):6475-81.
- Liu X, Kopparapu N-k, Li Y, Deng Y, Zheng X. Biochemical characterization of a novel fibrinolytic enzyme from Cordyceps militaris. Int J Biol Macromol. 2017;94:793-801.
- de Barros PDS, e Silva PEC, Nascimento TP, Costa RMPB, Bezerra RP, Porto ALF. Fibrinolytic enzyme from Arthrospira platensis cultivated in medium culture supplemented with corn steep liquor. Int J Biol Macromol. 2020;164:3446-53.
- Abdel-Fattah YR, Saeed HM, Gohar YM, El-Baz MA. Improved production of Pseudomonas aeruginosa uricase by optimization of process parameters through statistical experimental designs. Process Biochem. 2005;40(5):1707-14.
- Sharma KM, Kumar R, Panwar S, Kumar A. Microbial alkaline proteases: Optimization of production parameters and their properties. J Genet Eng Biotechnol. 2017;15(1):115-26.
- Taneja K, Bajaj BK, Kumar S, Dilbaghi N. Production, purification and characterization of fibrinolytic enzyme from Serratia sp. KG-2-1 using optimized media. Biotech. 2017;7(3):1-15.
- Devi CS, Mohanasrinivasan V, Sharma P, Das D, Vaishnavi B, Naine SJ. Production, purification and stability studies on nattokinase: a therapeutic protein extracted from mutant Pseudomonas aeruginosa CMSS isolated from bovine milk. Int J Pept Res Ther. 2016;22(2):263-9.
- Lee S-Y, Kim J-S, Kim J-E, Sapkota K, Shen M-H, Kim S, et al. Purification and characterization of fibrinolytic enzyme from cultured mycelia of Armillaria mellea. Protein Expr Purif. 2005;43(1):10-7.
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