Novel Sequential Screening and Enhanced Production of Fibrinolytic Enzyme byBacillussp. IND12 Using Response Surface Methodology in Solid-State Fermentation.
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,Bacillussp. IND12 was selected for fibrinolytic enzyme production.Bacillussp. IND12 effectively used cow dung for its growth and enzyme production (687 ± 6.5 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 MgSO4were the vital parameters with statistical significance (p< 0.001). 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 4143 ± 12.31 U/g material, which was more than fourfold the initial enzyme production (978 ± 36.4 U/g). The analysis of variance showed that the developed response surface model was highly significant (p< 0.001). The fibrinolytic enzyme digested goat blood clot (100%), chicken skin (83 ± 3.6%), egg white (100%), and bovine serum albumin (29 ± 4.9%).
Centre for Marine Science and Technology, Manonmaniam Sundaranar University, Rajakkamangalam, Kanyakumari District, Tamil Nadu 629 502, India.
Ponnuswamy
0000-0002-8083-2485
P
Vijayaraghavan
Kanyakumari Field Centre, Central Marine Fisheries Research Institute, Kanyakumari, Tamil Nadu 629702, India.
P
P
Rajendran
Centre for Marine Science and Technology, Manonmaniam Sundaranar University, Rajakkamangalam, Kanyakumari District, Tamil Nadu 629 502, India.
Samuel Gnana
0000-0002-6293-5544
SG
Prakash Vincent
Department of Biotechnology, Kalasalingam University, Srivilliputtur, Virudhunagar, Tamil Nadu 626126, India.
Arumugaperumal
0000-0001-5686-640X
A
Arun
Department of Botany and Microbiology, Addiriyah Chair for Environmental Studies, College of Science, King Saud University, P. O. Box 2455, Riyadh 11451, Saudi Arabia.
Naif
0000-0002-2663-6328
N
Abdullah Al-Dhabi
Department of Botany and Microbiology, Addiriyah Chair for Environmental Studies, College of Science, King Saud University, P. O. Box 2455, Riyadh 11451, Saudi Arabia.
Mariadhas
0000-0001-5898-2121
M
Valan Arasu
Department of Medical Education and Medical Humanities, School of Medicine, Kyung Hee University, Seoul, Republic of Korea.
Oh
O
Young Kwon
Department of Medicinal Crop Research, RDA, Eumseong, Chungbuk 369-873, Republic of Korea.
Young Ock
YO
Kim
D000818:Animals; D001407:Bacillus / Q000201:enzymology* / Q000254:growth & development*; D001426:Bacterial Proteins / Q000096:biosynthesis*; D002417:Cattle; D005343:Fibrinolytic Agents / Q000378:metabolism*; D006041:Goats; D010447:Peptide Hydrolases / Q000096:biosynthesis*
D001426:Bacterial Proteins; D005343:Fibrinolytic Agents; D010447:Peptide Hydrolases