基于数据库挖掘与蛋白质工程的PET降解酶研究进展
Research Progress on PET Degrading Enzymes Based on Database Mining and Protein Engineering
DOI: 10.12677/aep.2026.164069, PDF,   
作者: 王 剑：云南师范大学生命科学学院，西南联合研究生院，云南 昆明
关键词: 基因挖掘；聚对苯二甲酸乙二醇酯；竞争性抑制；理性设计；Gene Mining； Polyethylene Terephthalate； Competitive Inhibition； Rational Design
摘要: 聚对苯二甲酸乙二醇酯(PET)作为应用最广泛的合成塑料之一，其难降解特性引发的“白色污染”已成为全球性环境难题，生物降解法因绿色环保、条件温和成为PET回收的重要方向。PET水解酶的挖掘已形成天然环境筛选与数据库挖掘、宏基因组学等人工辅助技术并行的格局，筛选获得IsPETase、LCC等代表性酶类；通过理性设计、定向进化等蛋白质工程策略，可有效提升PET水解酶的催化活性、热稳定性等工业适配性能。同时，BHETase与MHETase可有效降解PET酶促降解中间产物，缓解其对PETase的竞争性抑制，与PET水解酶协同作用实现PET完全降解。本文综述了当前PET塑料的主流降解方法，重点聚焦生物降解法中PET水解酶的挖掘现状、蛋白质工程改造进展，以及PET酶促降解过程中关键中间产物(BHET, MHET)水解酶的研究情况，为高效、环保的PET生物降解技术开发提供关键酶资源和理论依据，对推动塑料污染治理及资源循环利用具有重要意义。
Abstract: Polyethylene terephthalate (PET) is one of the most widely used synthetic plastics, and the “white pollution” caused by its recalcitrant degradation has become a global environmental problem. Biodegradation has emerged as an important direction for PET recycling due to its environmental friendliness and mild reaction conditions. The mining of PET hydrolases has formed a pattern of parallel development of natural environment screening and artificial auxiliary technologies such as database mining and metagenomics, resulting in the screening of representative enzymes including IsPETase and LCC. Rational design, directed evolution and other protein engineering strategies can effectively improve the industrial adaptability of PET hydrolases, such as catalytic activity and thermal stability. Meanwhile, BHETase and MHETase can effectively degrade the key intermediate products of PET enzymatic degradation, alleviate their competitive inhibition on PETase, and achieve complete degradation of PET through synergistic action with PET hydrolases. This review summarizes the current mainstream degradation methods of PET plastics, focusing on the mining status of PET hydrolases in biodegradation, the progress of protein engineering modification, and the research status of hydrolases for key intermediate products (BHET, MHET) during PET enzymatic degradation. It provides key enzyme resources and theoretical basis for the development of efficient and environmentally friendly PET biodegradation technologies, and is of great significance for promoting plastic pollution control and resource recycling.
文章引用：王剑. 基于数据库挖掘与蛋白质工程的PET降解酶研究进展[J]. 环境保护前沿, 2026, 16(4): 691-699. https://doi.org/10.12677/aep.2026.164069

参考文献

[1] Geyer, R., Jambeck, J.R. and Law, K.L. (2017) Production, Use, and Fate of All Plastics Ever Made. Science Advances, 3, e1700782. [Google Scholar] [CrossRef] [PubMed]
[2] Shilpa, Basak, N. and Meena, S.S. (2022) Microbial Biodegradation of Plastics: Challenges, Opportunities, and a Critical Perspective. Frontiers of Environmental Science & Engineering, 16, Article No. 161. [Google Scholar] [CrossRef] [PubMed]
[3] Lebreton, L. and Andrady, A. (2019) Future Scenarios of Global Plastic Waste Generation and Disposal. Palgrave Communications, 5, Article No. 6. [Google Scholar] [CrossRef] 
[4] Pereyra-Camacho, M.A. and Pardo, I. (2024) Plastics and the Sustainable Development Goals: From Waste to Wealth with Microbial Recycling and Upcycling. Microbial Biotechnology, 17, e14459. [Google Scholar] [CrossRef] [PubMed]
[5] Wright, S.L. and Kelly, F.J. (2017) Plastic and Human Health: A Micro Issue? Environmental Science & Technology, 51, 6634-6647. [Google Scholar] [CrossRef] [PubMed]
[6] Thompson, R.C., Moore, C.J., vom Saal, F.S., et al. (2009) Plastics, the Environment and Human Health: Current Consensus and Future Trends. Philosophical Transactions of the Royal Society B, 364, 2153-2166. [Google Scholar] [CrossRef] [PubMed]
[7] Eriksen, M., Lebreton, L.C.M., Carson, H.S., Thiel, M., Moore, C.J., Borerro, J.C., et al. (2014) Plastic Pollution in the World’s Oceans: More than 5 Trillion Plastic Pieces Weighing over 250,000 Tons Afloat at Sea. PLOS ONE, 9, e111913. [Google Scholar] [CrossRef] [PubMed]
[8] Dhaka, V., Singh, S., Anil, A.G., Sunil Kumar Naik, T.S., Garg, S., Samuel, J., et al. (2022) Occurrence, Toxicity and Remediation of Polyethylene Terephthalate Plastics. a Review. Environmental Chemistry Letters, 20, 1777-1800. [Google Scholar] [CrossRef] [PubMed]
[9] Carr, C.M., Clarke, D.J. and Dobson, A.D.W. (2020) Microbial Polyethylene Terephthalate Hydrolases: Current and Future Perspectives. Frontiers in Microbiology, 11, Article ID: 571265. [Google Scholar] [CrossRef] [PubMed]
[10] Nguyen, H.T.H., Qi, P., Rostagno, M., Feteha, A. and Miller, S.A. (2018) The Quest for High Glass Transition Temperature Bioplastics. Journal of Materials Chemistry A, 6, 9298-9331. [Google Scholar] [CrossRef] 
[11] Rwei, S., Lin, W. and Wang, J. (2012) Synthesis and Characterization of Biodegradable and Weather-Durable PET/PEG/NDC Copolymers. Colloid and Polymer Science, 290, 1381-1392. [Google Scholar] [CrossRef] 
[12] Guo, Z., Wu, J. and Wang, J. (2025) Chemical Degradation and Recycling of Polyethylene Terephthalate (PET): A Review. RSC Sustainability, 3, 2111-2133. [Google Scholar] [CrossRef] 
[13] Soong, Y.V., Sobkowicz, M.J. and Xie, D. (2022) Recent Advances in Biological Recycling of Polyethylene Terephthalate (PET) Plastic Wastes. Bioengineering, 9, Article 98. [Google Scholar] [CrossRef] [PubMed]
[14] Garcia, J.M. and Robertson, M.L. (2017) The Future of Plastics Recycling. Science, 358, 870-872. [Google Scholar] [CrossRef] [PubMed]
[15] Ragaert, K., Delva, L. and Van Geem, K. (2017) Mechanical and Chemical Recycling of Solid Plastic Waste. Waste Management, 69, 24-58. [Google Scholar] [CrossRef] [PubMed]
[16] Siddiqui, M.N., Redhwi, H.H., Al-Arfaj, A.A. and Achilias, D.S. (2021) Chemical Recycling of PET in the Presence of the Bio-Based Polymers, PLA, PHB and PEF: A Review. Sustainability, 13, Article 10528. [Google Scholar] [CrossRef] 
[17] Cheng, L., Chen, X., Gu, J., Kobayashi, N., Yuan, H. and Chen, Y. (2025) Chemical Recycling of Waste Plastics: Current Challenges and Perspectives. Fundamental Research, 5, 919-922. [Google Scholar] [CrossRef] [PubMed]
[18] Wojnowska-Baryła, I., Bernat, K. and Zaborowska, M. (2022) Plastic Waste Degradation in Landfill Conditions: The Problem with Microplastics, and Their Direct and Indirect Environmental Effects. International Journal of Environmental Research and Public Health, 19, Article 13223. [Google Scholar] [CrossRef] [PubMed]
[19] 骆苑蓉, 钱义谦, 齐雅楠. 聚乙烯微塑料的微生物降解研究进展[J]. 环境科学, 2022, 43(11): 4869-4875.
[20] Austin, H.P., Allen, M.D., Donohoe, B.S., Rorrer, N.A., Kearns, F.L., Silveira, R.L., et al. (2018) Characterization and Engineering of a Plastic-Degrading Aromatic Polyesterase. Proceedings of the National Academy of Sciences, 115, E4350-E4357. [Google Scholar] [CrossRef] [PubMed]
[21] Yoshida, S., Hiraga, K., Takehana, T., Taniguchi, I., Yamaji, H., Maeda, Y., et al. (2016) A Bacterium That Degrades and Assimilates Poly(Ethylene Terephthalate). Science, 351, 1196-1199. [Google Scholar] [CrossRef] [PubMed]
[22] Danso, D., Chow, J. and Streit, W.R. (2019) Plastics: Environmental and Biotechnological Perspectives on Microbial Degradation. Applied and Environmental Microbiology, 85, e01095-19. [Google Scholar] [CrossRef] [PubMed]
[23] Liu, T., Xin, Y., Liu, X., et al. (2021) Advances in Microbial Degradation of Plastics. Journal of Bioengineering, 37, 2688-2702.
[24] Tournier, V., Topham, C.M., Gilles, A., David, B., Folgoas, C., Moya-Leclair, E., et al. (2020) An Engineered PET Depolymerase to Break down and Recycle Plastic Bottles. Nature, 580, 216-219. [Google Scholar] [CrossRef] [PubMed]
[25] Sonnendecker, C., Oeser, J., Richter, P.K., Hille, P., Zhao, Z., Fischer, C., et al. (2022) Low Carbon Footprint Recycling of Post‐Consumer PET Plastic with a Metagenomic Polyester Hydrolase. ChemSusChem, 15, e202101062. [Google Scholar] [CrossRef] [PubMed]
[26] Carniel, A., Gomes, A.d.C., Coelho, M.A.Z. and de Castro, A.M. (2021) Process Strategies to Improve Biocatalytic Depolymerization of Post-Consumer PET Packages in Bioreactors, and Investigation on Consumables Cost Reduction. Bioprocess and Biosystems Engineering, 44, 507-516. [Google Scholar] [CrossRef] [PubMed]
[27] Li, Q., Zheng, Y., Su, T., Wang, Q., Liang, Q., Zhang, Z., et al. (2022) Computational Design of a Cutinase for Plastic Biodegradation by Mining Molecular Dynamics Simulations Trajectories. Computational and Structural Biotechnology Journal, 20, 459-470. [Google Scholar] [CrossRef] [PubMed]
[28] Gao, R., Pan, H. and Lian, J. (2021) Recent Advances in the Discovery, Characterization, and Engineering of Poly(Ethylene Terephthalate) (PET) Hydrolases. Enzyme and Microbial Technology, 150, Article 109868. [Google Scholar] [CrossRef] [PubMed]
[29] Zhang, J., Shan, R., Li, X., et al. (2023) Enzymatic Properties and Degradation Characterization of a Bis(2-Hydroxyethyl) Terephthalate Hydrolase from Saccharothrix Sp. Chinese Journal of Biotechnology, 39, 2027-2039.
[30] Mrigwani, A., Pitaliya, M., Kaur, H., Kasilingam, B., Thakur, B. and Guptasarma, P. (2023) Rational Mutagenesis of thermobifida Fusca Cutinase to Modulate the Enzymatic Degradation of Polyethylene Terephthalate. Biotechnology and Bioengineering, 120, 674-686. [Google Scholar] [CrossRef] [PubMed]
[31] Thomsen, T.B., Radmer, T.S. and Meyer, A.S. (2024) Enzymatic Degradation of Poly(Ethylene Terephthalate) (PET): Identifying the Cause of the Hypersensitive Enzyme Kinetic Response to Increased PET Crystallinity. Enzyme and Microbial Technology, 173, Article 110353. [Google Scholar] [CrossRef] [PubMed]
[32] Kawai, F., Oda, M., Tamashiro, T., Waku, T., Tanaka, N., Yamamoto, M., et al. (2014) A Novel Ca2+-Activated, Thermostabilized Polyesterase Capable of Hydrolyzing Polyethylene Terephthalate from Saccharomonospora Viridis Ahk190. Applied Microbiology and Biotechnology, 98, 10053-10064. [Google Scholar] [CrossRef] [PubMed]
[33] Qiao, Y., Hu, R., Chen, D., Wang, L., Wang, Z., Yu, H., et al. (2022) Fluorescence-Activated Droplet Sorting of PET Degrading Microorganisms. Journal of Hazardous Materials, 424, 127417. [Google Scholar] [CrossRef] [PubMed]
[34] Handelsman, J. (2004) Metagenomics: Application of Genomics to Uncultured Microorganisms. Microbiology and Molecular Biology Reviews, 68, 669-685. [Google Scholar] [CrossRef] [PubMed]
[35] Zallot, R., Oberg, N. and Gerlt, J.A. (2019) The EFI Web Resource for Genomic Enzymology Tools: Leveraging Protein, Genome, and Metagenome Databases to Discover Novel Enzymes and Metabolic Pathways. Biochemistry, 58, 4169-4182. [Google Scholar] [CrossRef] [PubMed]
[36] Bateman, A., Martin, M.-J., Orchard, S. and Magrane, M. (2023) UniProt: the Universal Protein Knowledgebase in 2023. Nucleic Acids Research, 51, D523-D531.
[37] Baskaran, K. (2019) Protein Data Bank: The Single Global Archive for 3D Macromolecular Structure Data. Nucleic Acids Research, 47, D520-D528.
[38] Cooper, P.S., Lipshultz, D., Matten, W.T., McGinnis, S.D., Pechous, S., Romiti, M.L., et al. (2010) Education Resources of the National Center for Biotechnology Information. Briefings in Bioinformatics, 11, 563-569. [Google Scholar] [CrossRef] [PubMed]
[39] Hauenstein, J., Jeske, L., Jäde, A., Krull, M., Dümmer, K., Koblitz, J., et al. (2026) BRENDA in 2026: A Global Core Biodata Resource for Functional Enzyme and Metabolic Data within the DSMZ Digital Diversity. Nucleic Acids Research, 54, D527-D534. [Google Scholar] [CrossRef] 
[40] Mistry, J., Chuguransky, S., Williams, L., Qureshi, M., Salazar, G.A., Sonnhammer, E.L.L., et al. (2020) Pfam: The Protein Families Database in 2021. Nucleic Acids Research, 49, D412-D419. [Google Scholar] [CrossRef] [PubMed]
[41] Sulaiman, S., You, D., Kanaya, E., Koga, Y. and Kanaya, S. (2014) Crystal Structure and Thermodynamic and Kinetic Stability of Metagenome-Derived LC-Cutinase. Biochemistry, 53, 1858-1869. [Google Scholar] [CrossRef] [PubMed]
[42] Xi, X., Ni, K., Hao, H., Shang, Y., Zhao, B. and Qian, Z. (2021) Secretory Expression in Bacillus subtilis and Biochemical Characterization of a Highly Thermostable Polyethylene Terephthalate Hydrolase from Bacterium Hr29. Enzyme and Microbial Technology, 143, Article 109715. [Google Scholar] [CrossRef] [PubMed]
[43] Kato, S., Sakai, S., Hirai, M., Tasumi, E., Nishizawa, M., Suzuki, K., et al. (2018) Long-Term Cultivation and Metagenomics Reveal Ecophysiology of Previously Uncultivated Thermophiles Involved in Biogeochemical Nitrogen Cycle. Microbes and Environments, 33, 107-110. [Google Scholar] [CrossRef] [PubMed]
[44] Bornscheuer, U.T., Huisman, G.W., Kazlauskas, R.J., Lutz, S., Moore, J.C. and Robins, K. (2012) Engineering the Third Wave of Biocatalysis. Nature, 485, 185-194. [Google Scholar] [CrossRef] [PubMed]
[45] Son, H.F., Joo, S., Seo, H., Sagong, H., Lee, S.H., Hong, H., et al. (2020) Structural Bioinformatics-Based Protein Engineering of Thermo-Stable Petase from Ideonella Sakaiensis. Enzyme and Microbial Technology, 141, Article 109656. [Google Scholar] [CrossRef] [PubMed]
[46] Lee, S.H., Seo, H., Hong, H., Park, J., Ki, D., Kim, M., et al. (2023) Three-Directional Engineering of IsPETase with Enhanced Protein Yield, Activity, and Durability. Journal of Hazardous Materials, 459, Article 132297. [Google Scholar] [CrossRef] [PubMed]
[47] Lu, H., Diaz, D.J., Czarnecki, N.J., Zhu, C., Kim, W., Shroff, R., et al. (2022) Machine Learning-Aided Engineering of Hydrolases for PET Depolymerization. Nature, 604, 662-667. [Google Scholar] [CrossRef] [PubMed]
[48] Cui, Y., Chen, Y., Liu, X., Dong, S., Tian, Y., Qiao, Y., et al. (2021) Computational Redesign of a PETase for Plastic Biodegradation under Ambient Condition by the GRAPE Strategy. ACS Catalysis, 11, 1340-1350. [Google Scholar] [CrossRef] 
[49] Kawai, F. (2021) The Current State of Research on PET Hydrolyzing Enzymes Available for Biorecycling. Catalysts, 11, Article 206. [Google Scholar] [CrossRef] 
[50] Li, A., Sheng, Y., Cui, H., Wang, M., Wu, L., Song, Y., et al. (2023) Discovery and Mechanism-Guided Engineering of BHET Hydrolases for Improved PET Recycling and Upcycling. Nature Communications, 14, Article No. 4169. [Google Scholar] [CrossRef] [PubMed]
[51] Miao, R., Xu, G., Ding, Y., Ding, Z., Woodard, J., Tu, T., et al. (2024) Engineering Dual-Functional and Thermophilic BMHETase for Efficient Degradation of Polyethylene Terephthalate. Bioresource Technology, 414, Article 131556. [Google Scholar] [CrossRef] [PubMed]
[52] Palm, G.J., Reisky, L., Böttcher, D., Müller, H., Michels, E.A.P., Walczak, M.C., et al. (2019) Structure of the Plastic-Degrading Ideonella sakaiensis Mhetase Bound to a Substrate. Nature Communications, 10, Article No. 1717. [Google Scholar] [CrossRef] [PubMed]
[53] Knott, B.C., Erickson, E., Allen, M.D., Gado, J.E., Graham, R., Kearns, F.L., et al. (2020) Characterization and Engineering of a Two-Enzyme System for Plastics Depolymerization. Proceedings of the National Academy of Sciences, 117, 25476-25485. [Google Scholar] [CrossRef] [PubMed]
[54] von Haugwitz, G., Han, X., Pfaff, L., Li, Q., Wei, H., Gao, J., et al. (2022) Structural Insights into (Tere)Phthalate-Ester Hydrolysis by a Carboxylesterase and Its Role in Promoting PET Depolymerization. ACS Catalysis, 12, 15259-15270. [Google Scholar] [CrossRef] [PubMed]
[55] Erickson, E., Gado, J.E., Avilán, L., Bratti, F., Brizendine, R.K., Cox, P.A., et al. (2022) Sourcing Thermotolerant Poly(Ethylene Terephthalate) Hydrolase Scaffolds from Natural Diversity. Nature Communications, 13, Article No. 7850. [Google Scholar] [CrossRef] [PubMed]