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Keratinase production by Bacillus pumilus GHD in solid-state fermentation using sugar cane bagasse: optimisation of culture conditions using a Box-Behnken experimental design

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Abstract

Keratinase [EC 3.4.21/24/99.11] production by Bacillus pumilus GHD was investigated by solid-state fermentation (SSF) using sugar cane bagasse as a substrate. The SSF conditions were optimised using the “one variable at a time” strategy. A Box-Behnken design was employed to investigate the effects of adding the most common mineral salts on improving enzyme production. Maximal enzyme activity and amino acid production [730 ± 4.9 U/g dry solid (ds) and 368 ± 4.2 mg/g ds, respectively] was achieved using a combination of salts in concentrations of 0.087, 0.120, 0.204 g /10 g ds for NaCl, ZnCl2 and MgSO4, respectively. Capsicum annuum seeds were planted and irrigated with the amino-acid-rich culture filtrate of B. pumilus GHD. Growth rate, number of leaves and pigments (chlorophyll a and carotenoids) increased significantly with a 20-day irrigation at dilutions of 1:5 and 1:10 (v/v). The results of scanning electron microscopy showed the complete degradation of nail and horn upon treatment with the crude enzyme.

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Acknowledgement

This work was supported by National Research Centre, Chemistry of natural and microbial products department (Egypt).

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Authors and Affiliations

  1. Department of Chemistry of Microbial and Natural Products, National Research Centre, Tahrir Street, Dokki, Cairo, Egypt

    Ghada E. A. Awad, Mona A. Esawy, Walla A. Salam, Bassem M. Salama & Ahmed El-diwany

  2. Department of Plant Science, Faculty of Science, Ain Shams University, Abbassia, Cairo, Egypt

    Amal F. Abdelkader

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  1. Ghada E. A. Awad
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  2. Mona A. Esawy
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  3. Walla A. Salam
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  4. Bassem M. Salama
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  5. Amal F. Abdelkader
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  6. Ahmed El-diwany
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Correspondence to Ghada E. A. Awad.

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Awad, G.E.A., Esawy, M.A., Salam, W.A. et al. Keratinase production by Bacillus pumilus GHD in solid-state fermentation using sugar cane bagasse: optimisation of culture conditions using a Box-Behnken experimental design. Ann Microbiol 61, 663–672 (2011). https://doi.org/10.1007/s13213-010-0187-0

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