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Effect of bioprocess parameters on alkaline protease production by locally isolated Bacillus cereus AUST-7 using tannery waste in submerged fermentation

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Abstract

Present study aims to enhance protease production by employing locally isolated bacterial strains using animal skin (beef) wastes as a substrate. A proximate analysis of the substrate (skin waste) was carried out to know the percentage of protein, fats, and ash. To understand the particle size, shape, and other structural changes, scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FTIR) of both untreated and treated samples were performed. The substrate showed 83.81% crude protein, 11.23% crude fat, and 4.96% ash contents. SEM and FTIR analysis revealed changes in treated samples as compared to that untreated skin waste (control). In 140 isolated strains, 60 strains showed clear zone on 1% skim milk agar medium. The selected strains were cultured for protease assay on a production medium. The maximum protease (249.65 IU/ml) producing strain was identified as Bacillus cereus AUST-7 morphologically, biochemically, and based on 16S rDNA gene sequencing. The bioprocess parameters like aeration rate, shaking speed, and incubation period were optimized in lab scale bioreactor of 1.5 L capacity. During scale-up in a bioreactor (1.5 L), 2 volumes of aeration rate per volume of liquid medium per minute (VVM) aeration, shaking speed of 500 rpm, and 24 h of incubation period yielded maximum protease (1917.5 IU/mL) under submerged fermentation. The results depicted that defatted skin waste could be utilized as a potential substrate for the cost-effective production of alkaline protease.

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Authors and Affiliations

  1. Department of Microbiology, Abbottabad University of Science and Technology, Abbottabad, Pakistan

    Najeeb Ullah & Malik Mujaddad-ur-Rehman

  2. Food and Biotechnology Research Center, PCSIR Labs Complex, LahorePunjab, 54600, Pakistan

    Najeeb Ullah, Abid Sarwar, Muhammad Nadeem & Rubina Nelofer

  3. Department of Biotechnology, University of Sargodha, Sargodha, Pakistan

    Muhammad Irfan

  4. Department of Public Health, Government of Fujairah, Fujairah Municipality, Abattoir, United Arab Emirates

    Muhammad Idrees

  5. Department of Biotechnology, Quaid-E-Azam University, Islamabad, Pakistan

    Urooj Ali

  6. Department of Biochemistry, University of Malakand, Chakdara, 18800, Pakistan

    Sumaira Naz

  7. Department of Agriculture, University of Ioannina, Arta, Greece

    Tariq Aziz

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  1. Najeeb Ullah
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Contributions

Conceptualization, M.R and N.U.; methodology, N.U and M.N.; software, R.N.; validation, NU, A.S., S.N, M.Y.S, and U.A.; formal analysis, M.I.; investigation, M.I.D.; resources, T.A.; data curation, T.A.; writing—original draft preparation, N.U, S.N, and A.H.; writing—review and editing, T.A.; visualization, A.S and T.A.; supervision, M.I.R.; project administration, T.A.; funding acquisition, N.U. The manuscript was written through the contributions of all authors. All authors have approved the final version of the manuscript. All the authors contributed equally.

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Correspondence to Tariq Aziz.

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Ullah, N., Mujaddad-ur-Rehman, M., Sarwar, A. et al. Effect of bioprocess parameters on alkaline protease production by locally isolated Bacillus cereus AUST-7 using tannery waste in submerged fermentation. Biomass Conv. Bioref. (2023). https://doi.org/10.1007/s13399-023-04498-x

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