Development of hybrid biomicroparticles: cellulose exposing functionalized fusion proteins
Abstract Background One of the leading current trends in technology is the miniaturization of devices to the microscale and nanoscale. The highly advanced approaches are based on biological systems, subjected to bioengineering using chemical, enzymatic and recombinant methods. Here we have utilised...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-024-02344-x |
| _version_ | 1827315620893425664 |
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| author | Joanna Żebrowska Piotr Mucha Maciej Prusinowski Daria Krefft Agnieszka Żylicz-Stachula Milena Deptuła Aneta Skoniecka Agata Tymińska Małgorzata Zawrzykraj Jacek Zieliński Michał Pikuła Piotr M. Skowron |
| author_facet | Joanna Żebrowska Piotr Mucha Maciej Prusinowski Daria Krefft Agnieszka Żylicz-Stachula Milena Deptuła Aneta Skoniecka Agata Tymińska Małgorzata Zawrzykraj Jacek Zieliński Michał Pikuła Piotr M. Skowron |
| author_sort | Joanna Żebrowska |
| collection | DOAJ |
| description | Abstract Background One of the leading current trends in technology is the miniaturization of devices to the microscale and nanoscale. The highly advanced approaches are based on biological systems, subjected to bioengineering using chemical, enzymatic and recombinant methods. Here we have utilised the biological affinity towards cellulose of the cellulose binding domain (CBD) fused with recombinant proteins. Results Here we focused on fusions with ‘artificial’, concatemeric proteins with preprogrammed functions, constructed using DNA FACE™ technology. Such CBD fusions can be efficiently attached to micro-/nanocellulose to form functional, hybrid bionanoparticles. Microcellulose (MCC) particles were generated by a novel approach to enzymatic hydrolysis using Aspergillus sp. cellulase. The interaction between the constructs components – MCC, CBD and fused concatemeric proteins – was evaluated. Obtaining of hybrid biomicroparticles of a natural cellulose biocarrier with proteins with therapeutic properties, fused with CBD, was confirmed. Further, biological tests on the hybrid bioMCC particles confirmed the lack of their cytotoxicity on 46BR.1 N fibroblasts and human adipose derived stem cells (ASCs). The XTT analysis showed a slight inhibition of the proliferation of 46BR.1 N fibroblasts and ACSs cells stimulated with the hybrid biomicroparticles. However, in both cases no changes in the morphology of the examined cells after incubation with the hybrid biomicroparticles’ MCC were detected. Conclusions Microcellulose display with recombinant proteins involves utilizing cellulose, a natural polymer found in plants, as a platform for presenting or displaying proteins. This approach harnesses the structural properties of cellulose to express or exhibit various recombinant proteins on its surface. It offers a novel method for protein expression, presentation, or immobilization, enabling various applications in biotechnology, biomedicine, and other fields. Microcellulose shows promise in biomedical fields for wound healing materials, drug delivery systems, tissue engineering scaffolds, and as a component in bio-sensors due to its biocompatibility and structural properties. |
| first_indexed | 2024-04-24T23:01:45Z |
| format | Article |
| id | doaj.art-93e32ad629864aa3a5fcb4c23ed7ee18 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-24T23:01:45Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-93e32ad629864aa3a5fcb4c23ed7ee182024-03-17T12:42:51ZengBMCMicrobial Cell Factories1475-28592024-03-0123111810.1186/s12934-024-02344-xDevelopment of hybrid biomicroparticles: cellulose exposing functionalized fusion proteinsJoanna Żebrowska0Piotr Mucha1Maciej Prusinowski2Daria Krefft3Agnieszka Żylicz-Stachula4Milena Deptuła5Aneta Skoniecka6Agata Tymińska7Małgorzata Zawrzykraj8Jacek Zieliński9Michał Pikuła10Piotr M. Skowron11Department of Molecular Biotechnology, Faculty of Chemistry, University of GdanskDepartment of Molecular Biochemistry, Faculty of Chemistry, University of GdanskDepartment of Molecular Biotechnology, Faculty of Chemistry, University of GdanskDepartment of Molecular Biotechnology, Faculty of Chemistry, University of GdanskDepartment of Molecular Biotechnology, Faculty of Chemistry, University of GdanskLaboratory of Tissue Engineering and Regenerative Medicine, Division of Embryology, Faculty of Medicine, Medical University of GdanskLaboratory of Tissue Engineering and Regenerative Medicine, Division of Embryology, Faculty of Medicine, Medical University of GdanskLaboratory of Tissue Engineering and Regenerative Medicine, Division of Embryology, Faculty of Medicine, Medical University of GdanskDivision of Clinical Anatomy, Faculty of Medicine, Medical University of GdanskDepartment of Oncologic Surgery, Faculty of Medicine, Medical University of GdanskLaboratory of Tissue Engineering and Regenerative Medicine, Division of Embryology, Faculty of Medicine, Medical University of GdanskDepartment of Molecular Biotechnology, Faculty of Chemistry, University of GdanskAbstract Background One of the leading current trends in technology is the miniaturization of devices to the microscale and nanoscale. The highly advanced approaches are based on biological systems, subjected to bioengineering using chemical, enzymatic and recombinant methods. Here we have utilised the biological affinity towards cellulose of the cellulose binding domain (CBD) fused with recombinant proteins. Results Here we focused on fusions with ‘artificial’, concatemeric proteins with preprogrammed functions, constructed using DNA FACE™ technology. Such CBD fusions can be efficiently attached to micro-/nanocellulose to form functional, hybrid bionanoparticles. Microcellulose (MCC) particles were generated by a novel approach to enzymatic hydrolysis using Aspergillus sp. cellulase. The interaction between the constructs components – MCC, CBD and fused concatemeric proteins – was evaluated. Obtaining of hybrid biomicroparticles of a natural cellulose biocarrier with proteins with therapeutic properties, fused with CBD, was confirmed. Further, biological tests on the hybrid bioMCC particles confirmed the lack of their cytotoxicity on 46BR.1 N fibroblasts and human adipose derived stem cells (ASCs). The XTT analysis showed a slight inhibition of the proliferation of 46BR.1 N fibroblasts and ACSs cells stimulated with the hybrid biomicroparticles. However, in both cases no changes in the morphology of the examined cells after incubation with the hybrid biomicroparticles’ MCC were detected. Conclusions Microcellulose display with recombinant proteins involves utilizing cellulose, a natural polymer found in plants, as a platform for presenting or displaying proteins. This approach harnesses the structural properties of cellulose to express or exhibit various recombinant proteins on its surface. It offers a novel method for protein expression, presentation, or immobilization, enabling various applications in biotechnology, biomedicine, and other fields. Microcellulose shows promise in biomedical fields for wound healing materials, drug delivery systems, tissue engineering scaffolds, and as a component in bio-sensors due to its biocompatibility and structural properties.https://doi.org/10.1186/s12934-024-02344-xCellulose biomicroparticlesCellulose-binding-domain fusionsDNA-FACE technologyConcatemeric proteins |
| spellingShingle | Joanna Żebrowska Piotr Mucha Maciej Prusinowski Daria Krefft Agnieszka Żylicz-Stachula Milena Deptuła Aneta Skoniecka Agata Tymińska Małgorzata Zawrzykraj Jacek Zieliński Michał Pikuła Piotr M. Skowron Development of hybrid biomicroparticles: cellulose exposing functionalized fusion proteins Microbial Cell Factories Cellulose biomicroparticles Cellulose-binding-domain fusions DNA-FACE technology Concatemeric proteins |
| title | Development of hybrid biomicroparticles: cellulose exposing functionalized fusion proteins |
| title_full | Development of hybrid biomicroparticles: cellulose exposing functionalized fusion proteins |
| title_fullStr | Development of hybrid biomicroparticles: cellulose exposing functionalized fusion proteins |
| title_full_unstemmed | Development of hybrid biomicroparticles: cellulose exposing functionalized fusion proteins |
| title_short | Development of hybrid biomicroparticles: cellulose exposing functionalized fusion proteins |
| title_sort | development of hybrid biomicroparticles cellulose exposing functionalized fusion proteins |
| topic | Cellulose biomicroparticles Cellulose-binding-domain fusions DNA-FACE technology Concatemeric proteins |
| url | https://doi.org/10.1186/s12934-024-02344-x |
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