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Fabrication of Biofunctionalized Protease-Based Chitosan/Collagen Composite Membranes and Efficient Biodegradation Using Recombinant Aspergillus Fumigatus

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Abstract

The present study aimed at developing chitosan/collagen protease-based composite membranes loaded with enzyme nanoparticles and accessing its efficient biodegradation using a recombinantly produced aspartic protease from Aspergillus fumigatus (AfAP) with functional biochemical properties. By using Fourier transform infrared spectroscopy (FT-IR), X-ray diffractometry (XRD), and scanning electron microscopy (SEM) to analyze the resulting membranes, it was found that the ENPs and the chitosan-collagen composite membranes were highly compatible. Thermal properties (TG, DTG and DSC) were employed to evaluate the changes in the structural and physical properties which reached 384.94 °C, and a residual mass of 4.21 ± 0.13%. The ENPs loaded membranes exhibited marginally improved antioxidant and microbial growth inhibitory activities reaching 57.29 ± 2.77% and 10.33 ± 0.65 mm respectively. Additionally, the respective protease employed for the degradation studies was successfully expressed in Pichia pastoris (GS115) host cells, with functional pH and temperature optima between pH 4.0–5.0 and 60 °C respectively, and specific activity of 8408.9 ± 305.6 U/mg. It also exhibited enhanced specificity to its substrates upon collagen, chitosan, Bovine serum albumin (BSA), casein and casein sodium salt degradation with specific activities of 1477 ± 99.1, 1177 ± 103, and 1051.6 ± 60.7 U/mg respectively. Furthermore, the composite membranes showed excellent biodegradability by the 6th day in a natural environment treated with the aspartic protease from Aspergillus fumigatus (AfAP). Thus, the developed nanocomposite membranes from chitosan/collagen exhibited great potential and can well mimic the functions of a new generation of biodegradable and sustainable membranes.

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Funding

This study was financially supported by the Jiangsu Excellent Postdoctoral Project Fund (2022ZB681), the Jiangsu Agricultural Science and Technology Innovation Fund (CX(20)2029), the Key Research and Development Program (Modern Agriculture) of Jiangsu Province (BE2019358), and the Jiangsu University of Science and Technology Postdoctoral Research Fund (1062152103).

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Authors and Affiliations

  1. School of Materials Science and Engineering, Jiangsu University of Science and Technology, Zhenjiang, Jiangsu, 212100, P. R. China

    Richard A. Herman

  2. Jiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, Jiangsu, 212100, China

    Richard A. Herman, Xuan Zhu, Wen-Xin Zhang, Shuai You & Jun Wang

  3. College of Resources, Sichuan Agricultural University, Chengdu, Sichuan, 611130, P.R. China

    Ellen Ayepa

  4. School of Food and Biological Engineering, Jiangsu University, Zhenjiang, Jiangsu, 212013, P. R. China

    Michael Ackah

  5. Key Laboratory of Silkworm and Mulberry Genetic Improvement, Sericultural Research Institute, Ministry of Agricultural and Rural Affairs, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu, 212100, China

    Shuai You & Jun Wang

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Contributions

JW: Conceptualization, Fund acquisition, Supervision, Writing-review & editing, Project administration and Resources. SY: Conceptualization, Supervision, Writing-review & editing. RAH: Performed the major experiments, Data curation, Formal analysis, Writing – original draft, Writing – review & editing. ZX: Data curation. EA: Writing-review & editing. WXZ: Data curation. MA: Writing – review & editing. All authors reviewed the results and approved the final version of the manuscript.

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Herman, R.A., Zhu, X., Ayepa, E. et al. Fabrication of Biofunctionalized Protease-Based Chitosan/Collagen Composite Membranes and Efficient Biodegradation Using Recombinant Aspergillus Fumigatus. J Polym Environ 31, 3149–3166 (2023). https://doi.org/10.1007/s10924-023-02809-x

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