Biochemical Characterisation of Sis: A Distinct Thermophilic PETase with Enhanced NanoPET Substrate Hydrolysis and Thermal Stability
Abstract
:1. Introduction
2. Results and Discussion
2.1. Identification and Sequence Analysis of Sis
2.2. Determination of Optimal Reaction Conditions on pNP-Butyrate
2.3. Sis Can Degrade BHET and NanoPET at High Temperature
3. Materials and Methods
3.1. Identification, Sequence Analysis, and Structural Prediction of New Putative PET Hydrolysing Enzymes
3.2. Phylogenetic Analysis
3.3. Construction of Expression Vectors
3.4. Heterologous Expression and Purification
3.5. Differential Scanning Fluorimetry
3.6. Biochemical Characterisation on Pnp-Vutyrate
3.7. NanoPET Production
3.8. Enzymatic Degradation of NanoPET through PSP Assay
3.9. HPLC Analysis of Enzymatic Degradation of BHET and NanoPET
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Ercolano, C.; Iacono, R.; Cafaro, V.; Pizzo, E.; Giovannelli, D.; Feuerriegel, G.; Streit, W.R.; Strazzulli, A.; Moracci, M. Biochemical Characterisation of Sis: A Distinct Thermophilic PETase with Enhanced NanoPET Substrate Hydrolysis and Thermal Stability. Int. J. Mol. Sci. 2024, 25, 8120. https://doi.org/10.3390/ijms25158120
Ercolano C, Iacono R, Cafaro V, Pizzo E, Giovannelli D, Feuerriegel G, Streit WR, Strazzulli A, Moracci M. Biochemical Characterisation of Sis: A Distinct Thermophilic PETase with Enhanced NanoPET Substrate Hydrolysis and Thermal Stability. International Journal of Molecular Sciences. 2024; 25(15):8120. https://doi.org/10.3390/ijms25158120
Chicago/Turabian StyleErcolano, Carmen, Roberta Iacono, Valeria Cafaro, Elio Pizzo, Donato Giovannelli, Golo Feuerriegel, Wolfgang R. Streit, Andrea Strazzulli, and Marco Moracci. 2024. "Biochemical Characterisation of Sis: A Distinct Thermophilic PETase with Enhanced NanoPET Substrate Hydrolysis and Thermal Stability" International Journal of Molecular Sciences 25, no. 15: 8120. https://doi.org/10.3390/ijms25158120