Characterization of a PBAT Degradation Carboxylesterase from Thermobacillus composti KWC4
Abstract
:1. Introduction
2. Results
2.1. Protein Sequence Analysis
2.2. Characterization of Tcca
2.3. The Hydrolysis of PBAT Film by Tcca
2.4. Tcca Structure Modeling and Possible Substrate Binding Cavity
2.5. Protein Engineering of Tcca
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Cloning and Site−Directed Mutagenesis
4.3. Protein Purification
4.4. Characterization of Melting Temperature, Optimal pH and Temperature of Tcca
4.5. Scanning Electron Microscopy
4.6. Hydrolysis Activity of PBAT and BHET by Tcca and Its Mutants
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Residue Substitutions | Sequence (5′–3′) |
---|---|
Y77W | ATTGATGTTAATAATATT tgg ACCCGCGAATGGGCCGTG |
R79A | GTTAATAATATTTATACCgctGAATGGGCCGTGGACCCG |
E80A | AATAATATTTATACCCGCgctTGGGCCGTGGACCCGGAT |
W81A | AATATTTATACCCGCGAAgctGCCGTGGACCCGGATATT |
E129A | CAGGTGGGTCATGCAAGTgctATGGAATTTGATGGCGAA |
F130A | CATGCAAGTGAAATGGAAgctGATGGCGAACGCATTGCA |
F325A | GGTAATACCACCGATGAAgctATTAGTCGTCCGCAGGTT |
K427A | TTTTTTGAAACCTTAGCCgctTGTTGGCGCCCGTTTGTG |
W429A | GAAACCTTAGCCAAATGTgctCGCCCGTTTGTGGGCAAA |
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Wu, P.; Li, Z.; Gao, J.; Zhao, Y.; Wang, H.; Qin, H.; Gu, Q.; Wei, R.; Liu, W.; Han, X. Characterization of a PBAT Degradation Carboxylesterase from Thermobacillus composti KWC4. Catalysts 2023, 13, 340. https://doi.org/10.3390/catal13020340
Wu P, Li Z, Gao J, Zhao Y, Wang H, Qin H, Gu Q, Wei R, Liu W, Han X. Characterization of a PBAT Degradation Carboxylesterase from Thermobacillus composti KWC4. Catalysts. 2023; 13(2):340. https://doi.org/10.3390/catal13020340
Chicago/Turabian StyleWu, Pan, Zhishuai Li, Jian Gao, Yipei Zhao, Hao Wang, Huimin Qin, Qun Gu, Ren Wei, Weidong Liu, and Xu Han. 2023. "Characterization of a PBAT Degradation Carboxylesterase from Thermobacillus composti KWC4" Catalysts 13, no. 2: 340. https://doi.org/10.3390/catal13020340