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Optimization of media composition and growth conditions for production of milk-clotting protease (MCP) from Aspergillus oryzae DRDFS13 under solid-state fermentation

  • Biotechnology and Industrial Microbiology - Research Paper
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Abstract

This study reports the optimization of milk-clotting protease production from Aspergillus oryzae DRDFS13 under solid-state fermentation (SSF) in both one-variable-at-a-time and response surface methodology (RSM). The production and optimization of milk-clotting protease obtained from Aspergillus oryzae DRDFS13 under solid-state fermentation (SSF) using different agro-industrial wastes as solid substrates were studied. The agro-industrial wastes used included wheat bran, rice bran, pea bran, and grass pea bran. The chemical composition of the best solid substrate was tested using standard methods. Others cultivation parameters were studied, and the results showed that the optimum fermentation medium composed of wheat bran, casein (1% w/w), and glucose (0.5% w/w) and the conditions for maximum milk-clotting protease production were at the moisture content of 55.0%, inoculum of 0.5*106 spores/mL, incubation temperature of 30 °C, pH of 6.0, and fermentation time of 5 days. The highest milk-clotting activity was obtained from the crude enzyme extracted using 0.1 M NaCl and partial purification of the crude enzyme using chilled acetone, and 80% (NH4)2SO4 increased the ratio of MCA/PA from 0.56 to 1.30 and 0.65, respectively. Moreover, the highest MCA (137.58 U/mL) was obtained at a casein concentration of 0.5%, pH 4.0, and 25 °C, using RSM. Thus, results from the present study showed that the optimization of milk-clotting protease production from A. oryzae DRDFS 13 under SSF by both one-variable-at-a-time and RSM significantly increased the milk-clotting activity. This is the first report from a fungus in the Ethiopian setting and a modest contribution to highlight the potential of harnessing microbial protease enzymes for industrial applications.

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Acknowledgments

Moreover, we would like to acknowledge the Department of Microbial, Cellular, and Molecular Biology of Addis Ababa University and Downstream Processing Laboratory, Department of Life Sciences and Chemistry, Jacobs University, Bremen Germany for the facilitation of laboratory space and provision of basic facilities.

Funding

This work was supported by Microbial, Cellular, and Molecular Biology Department, Addis Ababa University.

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Authors and Affiliations

  1. Microbial, Cellular and Molecular Biology Department, Addis Ababa University, Addis Ababa, Ethiopia

    Jermen Mamo & Fassil Assefa

  2. Department of Biology, College of Natural and Computational Science, Debre Berhan University, Debre Berhan, Ethiopia

    Jermen Mamo

  3. Downstream Processing Laboratory, Department of Life Science and Chemistry, Jacobs University, Bremen, Germany

    Martin Kangwa & Hector Marcelo Fernandez-Lahore

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Mamo, J., Kangwa, M., Fernandez-Lahore, H.M. et al. Optimization of media composition and growth conditions for production of milk-clotting protease (MCP) from Aspergillus oryzae DRDFS13 under solid-state fermentation. Braz J Microbiol 51, 571–584 (2020). https://doi.org/10.1007/s42770-020-00243-y

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