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Kinetic and Parametric Optimization for the Enhanced Production of a Novel Fungal Exo-inulinase under Liquid Culture

Kinetic and Parametric Optimization for the Enhanced Production of a Novel Fungal Exo-inulinase under Liquid Culture

Sikander Ali* and S. Wajiha Khalid

Institute of Industrial Biotechnology, GC University, Lahore

* Corresponding author: [email protected]

ABSTRACT

The present work describes the optimization of process parameters for the enhanced production of a novel fungal exo-inulinase under liquid culture. Different fungal cultures were isolated and collected from various localities of Lahore and Murree (Punjab, Pakistan). Screening of these cultures was carried out in chemically defined medium i.e., 0.5 g/l (NH4)2SO4, 3 g/l KH2PO4, 1.5 g/l NaNO3, 0.01 g/l MgSO4.7H2O and 3 g/l inulin at pH 7. Kinetic parameters for comparing product yield coefficients of different fungal strains were applied to further optimize exo-inulinase production. The best enzyme producing culture was selected for optimization study. For this, the selected Rhizopus oligosporus (strain ROMII) was used for better enzyme production, for which another screening medium i.e., sucrose minerals salt at pH 5.5 was used. Different physical parameters were applied for maximum enzyme production. The enzyme exhibited maximum activity at pH 6 after 72 h of fermentation (158±0.39 U/ml). The partial purification of the enzyme was done with ammonium sulphate (20-80 %) followed by dialysis which resulted in 21 % purification from the crude enzyme extract. The molecular weight of enzyme was found to be 86 kDa, as determined by SDS poly-acrylamide gel electrophoresis. 

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Pakistan Journal of Zoology

October

Pakistan J. Zool., Vol. 56, Iss. 5, pp. 2001-2500

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