Fibrinolysin production by Alcaligenes faecalis strain 26 isolated from environment
Background and Objectives: Fibrinolytic drugs are commonly used for fibrin clot lysis but due to their inappropriate side effects, as well as their high costs, using fibrinolytic enzymes has been paid attention. Bacterial sources of this enzyme are a good alternative for this purpose. The aim was fibrinolysin production through screening of fibrinolysin producing bacteria from environmental samples.
Materials and Methods: Bacterial isolation was performed from different environmental samples and was screened based on sheep blood clot digestion and culture on plasma plate. The most potent isolate was optimized for different growth parameters including temperature, pH and fibrinolysin production at optimum growth conditions. The stability of produced enzyme at various temperatures and pH and treatment with MgSO4, NiSO4, SDS and EDTA was then investigated. Finally this isolate was identified based on the 16S rRNA sequencing.
Results: As a result, from 79 different isolates, the most potent fibrinolysin producer was identified as Alcaligenes faecalis strain 26. This isolate produced 12 mm halo zone on plasma plate. Its optimum growth temperature and pH was 43°C and 7, respectively. The produced enzyme had the best stability at pH 7 and was also active up to 60°C. The fibrinolytic activity of this isolate was reduced following treatment with MgSO4, NiSO4 and also with protease inhibitors, such as SDS and EDTA.
Conclusion: Based on the obtained results it can be suggested that Alcaligenes faecalis strain 26 has appropriate efficiency for fibrinolysin production that can be used in food industry and medicine.
Jayalakshmi T, Krishnamoorthy P, Ramesh Babu PB, Vidhya B. Production, purification and biochemical characterization of alkaline fibrinolytic enzyme from Bacillus subtilis¬ strain-GBRC1. J Chem Pharm Res 2012; 4: 5027-5031.
Liu XL, Zheng XQ, Qian PZ, Kopparapu NK, Deng YP, Nonaka M. Purification and chracterization of a novel fibrinolytic enzyme from culture supernatant of Pleurotus ostreaus. J Microbiol Biotechnol 2014; 24: 245-253.
Thokchom S, Joshi SR. Screening of fibrinolytic enzymes from lactic acid bacterial isolates associated with traditional fermented soyben foods. Food Sci Biotechnol 2014; 23: 1601-1604.
Venkatanagaraju E, Divakar G. An overview on microbial fibrinolytic protease. Int J Pharm Sci Res 2012; 5: 643-656.
Yang J, Yang Ji, Zhuang Z, Yang Y, Lin L, Wang S. Thrombolytic effects of Douchi fibrinolytic from Bacillus subtilis LD-8547 in vitro and in vivo. Biomed Central Biotechnol 2012; 12: 1-36.
Spijker T, Graaff R, Boonstra PW, Busscher HJ, Van Oeveren W. On the influence of flow conditions and wettability on blood material interactions. Biomaterials 2003; 24: 4717-4727.
Shadan F (1996). Medical physiology. In: Hemostasis. Ed, AC Guyton, JE Hall. Chehr Joint-stock Company publishing, Tehran, pp. 717-729.
Tengborn L. Fibrinolytic inhibitor in the management of bleeding disorder. Hemophilia 2012; 42: 1-14.
Chen B, Huo J, He Z, He Q, Hao Y, Chen Z. Isolation and identification of an efective fibrinolytic strain Bacillus subtilis FR-33 from the Chinese doufuru and primary analysis of its fibrinolytic enzyme. Afr J Microbiol Res 2013; 7: 2001-2009.
Venkatanagaraju E, Divakar G. Bacillus cereus GD 55 strain improvement by physical and chemical mutagenesis for enhanced production of fibrinolytic protease. Int J Pharm Sci Res 2013; 4: 81-93.
Hwang KJ, Choi KH, Kim MJ, Park CS, Cha G. Purification and Characterization of a new fibrinolytic enzyme of Bacillus licheniformis KJ-31, isolated from Korean traditional Jeot-gal. J Microbiol Biotechnol 2007; 17: 1469-1476.
Ho Ko J, Peng Yan J, Zhu L, Peng Qi Y. Identification of two novel fibrinolytic enzymes from Bacillus subtilis QK02. Comp Biochem Physiol 2004; 137: 65-74.
Latridis SG, Ferguson JH. The plasma plate method for estimating thrombolytic activity. Exp Biol Med 2015; 110: 738-740.
Lee SK, Bae DH, Kwon TJ, Lee SB, Lee HH, Park JH. Purification and characterization of a fibrinolytic enzyme from Bacillus sp. KDO-13 isolated from soybean paste. J Microbiol Biotechnol 2001; 11: 845-852.
Ausubel FM, Brent R, Kingstone RE, Moor DD, Seidman JG, Smith JA, Struhl K (1992). Short protocols in molecular biology. 2nd ed. New York.
Krik O, Borchet TV, Fuglsang CC. Industrial enzyme applications. Curr Opin Biotechnol 2002; 13: 345-351.
Cheng G, He L, Sun Z, Cui Z, Du Y, Kong Y. Purification and biochemical characterization of a novel fibrinolytic enzyme from Streptomyces sp. P3. J Microbiol Biothecnol 2015; 25: 1449-1459.
Jo HD, Lee HA, Jeong SJ, Kim JH. Purification and characterization of a major fibrinolytic enzyme from Bacillus amyloliquefaciens MJ5-41 isolated from Meju. J Microbiol Biotechnol 2011; 21: 1166-1173.
Kim WK, Choi KH, Kim YT, Park HH, Choi JY, Le YS. Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. Strain CK11-4 screened from Chungkookjang. Appl Environ Microbiol 1996; 62: 2482-2488.