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BioMed Research International
Volume 2017, Article ID 3909657, 13 pages
Research Article

Novel Sequential Screening and Enhanced Production of Fibrinolytic Enzyme by Bacillus sp. IND12 Using Response Surface Methodology in Solid-State Fermentation

1Centre for Marine Science and Technology, Manonmaniam Sundaranar University, Rajakkamangalam, Kanyakumari District, Tamil Nadu 629 502, India
2Kanyakumari Field Centre, Central Marine Fisheries Research Institute, Kanyakumari, Tamil Nadu 629702, India
3Department of Biotechnology, Kalasalingam University, Srivilliputtur, Virudhunagar, Tamil Nadu 626126, India
4Department of Botany and Microbiology, Addiriyah Chair for Environmental Studies, College of Science, King Saud University, P. O. Box 2455, Riyadh 11451, Saudi Arabia
5Department of Medical Education and Medical Humanities, School of Medicine, Kyung Hee University, Seoul, Republic of Korea
6Department of Medicinal Crop Research, RDA, Eumseong, Chungbuk 369-873, Republic of Korea

Correspondence should be addressed to Ponnuswamy Vijayaraghavan; moc.liamg@semyznev and Mariadhas Valan Arasu; moc.liamg@usaranalavm

Received 8 August 2016; Revised 30 November 2016; Accepted 13 December 2016; Published 22 February 2017

Academic Editor: Denise Freire

Copyright © 2017 Ponnuswamy Vijayaraghavan et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Fibrinolytic enzymes have wide applications in clinical and waste treatment. Bacterial isolates were screened for fibrinolytic enzyme producing ability by skimmed milk agar plate using bromocresol green dye, fibrin plate method, zymography analysis, and goat blood clot lysis. After these sequential screenings, Bacillus sp. IND12 was selected for fibrinolytic enzyme production. Bacillus sp. IND12 effectively used cow dung for its growth and enzyme production ( U/g substrate). Further, the optimum bioprocess parameters were found out for maximum fibrinolytic enzyme production using cow dung as a low cost substrate under solid-state fermentation. Two-level full-factorial experiments revealed that moisture, pH, sucrose, peptone, and MgSO4 were the vital parameters with statistical significance (). Three factors (moisture, sucrose, and MgSO4) were further studied through experiments of central composite rotational design and response surface methodology. Enzyme production of optimized medium showed  U/g material, which was more than fourfold the initial enzyme production ( U/g). The analysis of variance showed that the developed response surface model was highly significant (). The fibrinolytic enzyme digested goat blood clot (100%), chicken skin (%), egg white (100%), and bovine serum albumin (%).