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Production, purification and characterization of fibrinolytic enzyme from Serratia sp. KG-2-1 using optimized media

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Abstract

Intravascular thrombosis is one of the major causes of variety of cardiovascular disorders leading to high mortality worldwide. Fibrinolytic enzymes from microbial sources possess ability to dissolve these clots and help to circumvent these problems in more efficient and safer way. In the present study, fibrinolytic protease with higher fibrinolytic activity than plasmin was obtained from Serratia sp. KG-2-1 isolated from garbage dump soil. Response surface methodology was used to study the interactive effect of concentration of maltose, yeast extract + peptone (1:1), incubation time, and pH on enzyme production and biomass. Maximum enzyme production was achieved at 33 °C after 24 h at neutral pH in media containing 1.5% Maltose, 4.0% yeast extract + peptone and other trace elements resulting in 1.82 folds increased production. The enzyme was purified from crude extract using ammonium sulfate precipitation and DEAE-Sephadex chromatography resulting in 12.9 fold purification with 14.9% yield. The purified enzyme belongs to metalloprotease class and had optimal activity in conditions similar to physiological environment with temperature optima of 40 °C and pH optima of 8. The enzyme was found to be stable in various solvents and its activity was enhanced in presence of Na+, K+, Ba2+, Cu2+, Mn2+, Hg2+ but inhibited by Ca2+ and Fe3+. Hence, the obtained enzyme may be used as potential therapeutic agent in combating various thrombolytic disorders.

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Acknowledgements

The author Kapila Taneja gratefully acknowledges Department of Science and Technology, Govt. of India for providing financial support in the form of DST-INSPIRE Fellowship. Authors also thank Microbial Culture Collection, NCCS, DBT, Govt. of India for phenotypic characterization of samples.

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Authors and Affiliations

  1. Department of Bio and Nano Technology, Guru Jambheshwar University of Science and Technology, Hisar, Haryana, 125001, India

    Kapila Taneja, Sandeep Kumar & Neeraj Dilbaghi

  2. School of Biotechnology, University of Jammu, Jammu, 180006, India

    Bijender Kumar Bajaj

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  1. Kapila Taneja
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  2. Bijender Kumar Bajaj
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  3. Sandeep Kumar
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  4. Neeraj Dilbaghi
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Correspondence to Neeraj Dilbaghi.

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Taneja, K., Bajaj, B.K., Kumar, S. et al. Production, purification and characterization of fibrinolytic enzyme from Serratia sp. KG-2-1 using optimized media. 3 Biotech 7, 184 (2017). https://doi.org/10.1007/s13205-017-0808-4

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