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Heterologous expression and purification of keratinase from Actinomadura viridilutea DZ50: feather biodegradation and animal hide dehairing bioprocesses

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Abstract

The keratin-degrading bacterium Actinomadura viridilutea DZ50 secretes a keratinase (KERDZ) with potential industrial interest. Here, the kerDZ gene was extracellularly expressed in Escherichia coli BL21(DE3)pLysS using pTrc99A vector. The recombinant enzyme (rKERDZ) was purified and biochemically characterized. Results showed that the native and recombinant keratinases have similar biochemical characteristics. The conventional dehairing with lime and sodium sulfide degrades the hair to the extent that it cannot be recovered. Thus, these chemical processes become a major contributor to wastewater problem and create a lot of environmental concern. The complete dehairing was achieved with 2000 U/mL rKERDZ for 10 h at 40 °C. In fact, keratinase assisted dehairing entirely degraded chicken feather (45 mg) and removed wool/hair from rabbit, sheep, goat, or bovine’ hides (1.6 kg) while preserving the collagen structure. The enzymatic process is the eco-friendly option that reduces biological (BOD) (50%) and chemical (COD) oxygen demands (60%) in leather processing. Consequently, the enzymatic hair removal process could solve the problem of post-treatments encountering the traditional leather processing. The enzymatic (rKERDZ) dehaired leather was analyzed by scanning electron microscopic (SEM) studies, which revealed similar fiber orientation and compactness compared with control sample. Those properties support that the rKERDZ enzyme–mediated process is greener to some extent than the traditional one.

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Acknowledgments

The authors would like to express their gratitude to Mr. K. Walha, Mrs. N. Kchaou, and Mrs N. Masmoudi (Analysis Unit-CBS), Mr. I. Hsairi, Mr. A. Zitoun, and Mr. F. Boukhili (UVRR-CBS) for their technical assistance. We thank also our industrial partners, Mr. E. Ben Moussa, Mr. M. S. Bechir, and Mr. M. Hmila from “SO.SA.CUIR” leather tannery (M’Saken, Sousse, Tunisia), for granting us access to their technical facility and kindly donating the leather batting enzymes used in this study. Many thanks are also owed to Pr. M. Hammami and Mr. M. Marzouki (INRAP, Technopole Sidi Thabet, Ariana, Tunisia) for their help with the scanning electron microscopy (SEM). Special thanks are also due to Prof. C. Khalaf, Senior ESL/EFL Instructor, Translator, and Proofreader from the Faculty of Medicine of Sfax, University of Sfax (Sfax, Tunisia); Dr. Z. Bouallagui from the LBPE-CBS; Pr. A. Bouanane-Darenfed from the LCBM-FSB/USTHB (Algiers, Algeria); and Dr. A. Noiriel and Pr. A. Abousalham from the Université Lyon 1, ICBMS, UMR 5246 CNRS-MEM2 (Lyon, France) for their constructive proofreading and language polishing services.

Funding

This work was funded by the Tunisian Ministry of Higher Education and Scientific Research in the framework of the Contract Programs (LBMIE-CBS, code grant no.: LR15CBS06 2015-2018 and LBMEB-CBS, code grant no.: LR19CBS05 2019-2022), the multilateral project Partenariats Hubert Curien (PHC)-Maghreb 2020 Program (FranMaghZYM 2020-2023, code Campus France: 43791TM, code PHC: 01MAG20), the Tunisian Ministry of Industry “National Programme for the Promotion of Technological Innovation, grant no. PNRI-Dégraissage-Reverdissage 2016–2020,” the Tunisian–Algerian project JAOUADI/BADIS_TNDZ-MicrooZymes_2012-2018/code: TA/04/2012, and the PAQ-Collabora_1/2018 (PAR&I-Tk) Biotech-INNOV/CBS 2018–2022.

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Authors and Affiliations

  1. Laboratory of Microbial Biotechnology, Enzymatic, and Biomolecules (LMBEB), Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, 3018, Sfax, Tunisia

    Mouna Ben Elhoul, Nadia Zaraî Jaouadi, Hatem Rekik, Sondes Mechri & Bassem Jaouadi

  2. Biotech ECOZYM Start-up, Business Incubator, Centre of Biotechnology of Sfax (CBS), University of Sfax, Road of Sidi Mansour Km 6, P.O. Box 1177, 3018, Sfax, Tunisia

    Mouna Ben Elhoul, Nadia Zaraî Jaouadi, Hatem Rekik & Bassem Jaouadi

  3. Laboratory of Cellular and Molecular Biology (LCMB), Microbiology Team, Faculty of Biological Sciences, University of Sciences and Technology of Houari Boumediene (USTHB), P.O. Box 32, El Alia, Bab Ezzouar, 16111, Algiers, Algeria

    Khelifa Bouacem & Fawzi Allala

  4. National Leather and Shoe Center (CNCC), 17, Road of leather, Z.I. Sidi Rezig, 2033, Ben Arous, Tunisia

    Haifa Khemir Ezzine & Neila Miled

Authors
  1. Mouna Ben Elhoul
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  2. Nadia Zaraî Jaouadi
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  3. Khelifa Bouacem
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  4. Fawzi Allala
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  5. Hatem Rekik
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  6. Sondes Mechri
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  7. Haifa Khemir Ezzine
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  8. Neila Miled
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  9. Bassem Jaouadi
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Contributions

MBE and NZJ carried out the experiment. MBE wrote the manuscript with support from BJ, SM, HR, and KB. FA aided in interpreting the results and worked on the manuscript. HKE and NM manufactured the samples and characterized them by scanning electron microscopy (SEM). All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Bassem Jaouadi.

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Ben Elhoul, M., Zaraî Jaouadi, N., Bouacem, K. et al. Heterologous expression and purification of keratinase from Actinomadura viridilutea DZ50: feather biodegradation and animal hide dehairing bioprocesses. Environ Sci Pollut Res 28, 9921–9934 (2021). https://doi.org/10.1007/s11356-020-11371-1

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