Optimization of keratinase production and enzyme activity using response surface methodology with streptomyces sp7 |
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Authors: | Radhika Tatineni Kiran Kumar Doddapaneni Ravi Chandra Potumarthi Lakshmi Narasu Mangamoori |
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Institution: | (1) Center for Biotechnology, Institute of Science and Technology, JNT University, 500085 Hyderabad, India;(2) Biochemical and Environmental Engineering Center, Indian Institute of Chemical Technology, Hyderabad |
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Abstract: | A two-step response surface methodology (RSM) study was conducted for the optimization of keratinase production and enzyme
activity from poultry feather byStreptomyces sp7. Initially different combinations of salts were screened for maximal production of keratinase at a constant pH of 6.5
and feather meal concentration of 5 g/L. A combination of K2HPO4, KH2PO4, and NaCl gave a maximum yield of keratinase (70.9 U/mL) production. In the first step of the RSM study, the selected five
variables (feather meal, K2HPO4, KH2PO4, NaCl, and pH) were optimized by a 25 full-factorial rotatable central composite design (CCD) that resulted in 95 U/mL of keratinase production. The results of
analysis of variance and regression of a second-order model showed that the linear effects of feather meal concentration (p<0.005) and NaCl (p<0.029) and the interactive effects of all variables were more significant and that values of the quadratic effects of feather
meal (p<1.72e-5), K2HPO4 (p<4.731e-6), KH2PO4 (p<1.01e-10), and pH (p 7.63e-7) were more significant than the linear and interactive effects of the process variables. In the second step, a 23 rotatable full-factorial CCD and response surface analysis were used for the selection of optimal process parameters (pH,
temperature, and rpm) for keratinase enzyme activity. These optima were pH 11.0, 45°C, and 300 rpm. |
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Keywords: | Poultry feather keratinase response surface methodology central composite design |
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