Characterization of a PBAT Degradation Carboxylesterase from <i>Thermobacillus composti</i> KWC4
The large amount of waste synthetic polyester plastics has complicated waste management and also endangering the environment due to improper littering. In this study, a novel carboxylesterase from <i>Thermobacillus composti</i> KWC4 (Tcca) was identified, heterologously expressed in <...
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MDPI AG
2023-02-01
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| Series: | Catalysts |
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| Online Access: | https://www.mdpi.com/2073-4344/13/2/340 |
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| author | Pan Wu Zhishuai Li Jian Gao Yipei Zhao Hao Wang Huimin Qin Qun Gu Ren Wei Weidong Liu Xu Han |
| author_facet | Pan Wu Zhishuai Li Jian Gao Yipei Zhao Hao Wang Huimin Qin Qun Gu Ren Wei Weidong Liu Xu Han |
| author_sort | Pan Wu |
| collection | DOAJ |
| description | The large amount of waste synthetic polyester plastics has complicated waste management and also endangering the environment due to improper littering. In this study, a novel carboxylesterase from <i>Thermobacillus composti</i> KWC4 (Tcca) was identified, heterologously expressed in <i>Escherichia coli</i>, purified and characterized with various plastic substrates. Irregular grooves were detected on polybutylene adipate terephthalate (PBAT) film by scanning electron microscopy (SEM) after Tcca treatment, and Tcca can also hydrolyze short–chain diester bis(hydroxyethyl) terephthalate (BHET). The optimal pH and temperature for Tcca were 7.0 and 40 °C, respectively. In order to explore its catalytic mechanism and improve its potential for plastic hydrolysis, we modeled the protein structure of Tcca and compared it with its homologous structures, and we identified positions that might be crucial for the binding of substrates. We generated a variety of Tcca variants by mutating these key positions; the variant F325A exhibited a more than 1.4–fold improvement in PBAT hydrolytic activity, and E80A exhibited a more than 4.1–fold increase in BHET activity when compared to the wild type. Tcca and its variants demonstrated future applicability for the recycling of bioplastic waste containing a PBAT fraction. |
| first_indexed | 2024-03-11T09:01:46Z |
| format | Article |
| id | doaj.art-3bc664b588784475b8651eb9d5096073 |
| institution | Directory Open Access Journal |
| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-11T09:01:46Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
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| spelling | doaj.art-3bc664b588784475b8651eb9d50960732023-11-16T19:41:46ZengMDPI AGCatalysts2073-43442023-02-0113234010.3390/catal13020340Characterization of a PBAT Degradation Carboxylesterase from <i>Thermobacillus composti</i> KWC4Pan Wu0Zhishuai Li1Jian Gao2Yipei Zhao3Hao Wang4Huimin Qin5Qun Gu6Ren Wei7Weidong Liu8Xu Han9College of Biotechnology, Tianjin University of Science and Technology, No. 9, 13th Avenue, Tianjin Economic and Technological Development Area, Tianjin 300457, ChinaTianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin 300308, ChinaTianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin 300308, ChinaCollege of Biotechnology, Tianjin University of Science and Technology, No. 9, 13th Avenue, Tianjin Economic and Technological Development Area, Tianjin 300457, ChinaTianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin 300308, ChinaCollege of Biotechnology, Tianjin University of Science and Technology, No. 9, 13th Avenue, Tianjin Economic and Technological Development Area, Tianjin 300457, ChinaTianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin 300308, ChinaJunior Research Group Plastic Biodegradation, Institute of Biochemistry, University of Greifswald, Felix–Hausdorf–Str. 8, 17489 Greifswald, GermanyTianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin 300308, ChinaTianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin 300308, ChinaThe large amount of waste synthetic polyester plastics has complicated waste management and also endangering the environment due to improper littering. In this study, a novel carboxylesterase from <i>Thermobacillus composti</i> KWC4 (Tcca) was identified, heterologously expressed in <i>Escherichia coli</i>, purified and characterized with various plastic substrates. Irregular grooves were detected on polybutylene adipate terephthalate (PBAT) film by scanning electron microscopy (SEM) after Tcca treatment, and Tcca can also hydrolyze short–chain diester bis(hydroxyethyl) terephthalate (BHET). The optimal pH and temperature for Tcca were 7.0 and 40 °C, respectively. In order to explore its catalytic mechanism and improve its potential for plastic hydrolysis, we modeled the protein structure of Tcca and compared it with its homologous structures, and we identified positions that might be crucial for the binding of substrates. We generated a variety of Tcca variants by mutating these key positions; the variant F325A exhibited a more than 1.4–fold improvement in PBAT hydrolytic activity, and E80A exhibited a more than 4.1–fold increase in BHET activity when compared to the wild type. Tcca and its variants demonstrated future applicability for the recycling of bioplastic waste containing a PBAT fraction.https://www.mdpi.com/2073-4344/13/2/340polybutylene adipate terephthalate (PBAT)carboxyl esteraseenzymatic degradationprotein engineering |
| spellingShingle | Pan Wu Zhishuai Li Jian Gao Yipei Zhao Hao Wang Huimin Qin Qun Gu Ren Wei Weidong Liu Xu Han Characterization of a PBAT Degradation Carboxylesterase from <i>Thermobacillus composti</i> KWC4 Catalysts polybutylene adipate terephthalate (PBAT) carboxyl esterase enzymatic degradation protein engineering |
| title | Characterization of a PBAT Degradation Carboxylesterase from <i>Thermobacillus composti</i> KWC4 |
| title_full | Characterization of a PBAT Degradation Carboxylesterase from <i>Thermobacillus composti</i> KWC4 |
| title_fullStr | Characterization of a PBAT Degradation Carboxylesterase from <i>Thermobacillus composti</i> KWC4 |
| title_full_unstemmed | Characterization of a PBAT Degradation Carboxylesterase from <i>Thermobacillus composti</i> KWC4 |
| title_short | Characterization of a PBAT Degradation Carboxylesterase from <i>Thermobacillus composti</i> KWC4 |
| title_sort | characterization of a pbat degradation carboxylesterase from i thermobacillus composti i kwc4 |
| topic | polybutylene adipate terephthalate (PBAT) carboxyl esterase enzymatic degradation protein engineering |
| url | https://www.mdpi.com/2073-4344/13/2/340 |
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