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African Journal of Biotechnology

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Optimization of extracellular catalase production from Aspergillus phoenicis K30 by a linear regression method using date flour as single carbon source and purification of the enzyme

N Chaouche Kacem, J Destain, Z Meraihi, L Dehimat, T Haddoum, JP Wathelet, Ph Thonart

Abstract


Aspergillus phoenicis K30 is the selected mutant which produces an amount of extracellular catalase. To amplify the extracellular catalase production by the strain, a fermentation optimization was performed. To select the factors affecting the production, nine active variables (factors) consisting of 12 experiments were analyzed by Plackett-Burman design. Each variable was tested at two levels, a higher and a lower level. The studies of the effect of each variable and the establishment of a correlation between the response of enzyme activity and variables revealed that the link is a multiple linear regression form. The optimization was carried out through a simplex algorithm. The amount of extracellular catalase produced by the strain in the optimized medium was about four times higher than that obtained in non optimized medium corresponding to 3820 mg/L of extracellular proteins including 59500 U/L of extracellular catalase activity after 96 h of fermentation. The steps of purification were allowed to improve enzyme activity by 305-fold. From an analytical gel electrophoresis under native conditions, an apparent molecular mass of 158 kDa was determined suggesting that the enzyme is a homodimer. The isoelectric point of the protein was found to be 5 ± 0.1 as determined by a Pharmacia Phast-system.

Keywords: Aspergillus phoenicis, extracellular catalase purification, dates flour, optimization, multiple linear regression.

African Journal of Biotechnology Vol. 12(19), pp. 2646-2653



AJOL African Journals Online