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Discovery and rational engineering of PET hydrolase with both mesophilic and thermophilic PET hydrolase properties

Author

Listed:
  • Hwaseok Hong

    (Kyungpook National University)

  • Dongwoo Ki

    (Kyungpook National University)

  • Hogyun Seo

    (Kyungpook National University)

  • Jiyoung Park

    (Kyungpook National University)

  • Jaewon Jang

    (CJ CheilJedang Co.)

  • Kyung-Jin Kim

    (Kyungpook National University
    Zyen Co)

Abstract

Excessive polyethylene terephthalate (PET) waste causes a variety of problems. Extensive research focused on the development of superior PET hydrolases for PET biorecycling has been conducted. However, template enzymes employed in enzyme engineering mainly focused on IsPETase and leaf-branch compost cutinase, which exhibit mesophilic and thermophilic hydrolytic properties, respectively. Herein, we report a PET hydrolase from Cryptosporangium aurantiacum (CaPETase) that exhibits high thermostability and remarkable PET degradation activity at ambient temperatures. We uncover the crystal structure of CaPETase, which displays a distinct backbone conformation at the active site and residues forming the substrate binding cleft, compared with other PET hydrolases. We further develop a CaPETaseM9 variant that exhibits robust thermostability with a Tm of 83.2 °C and 41.7-fold enhanced PET hydrolytic activity at 60 °C compared with CaPETaseWT. CaPETaseM9 almost completely decompose both transparent and colored post-consumer PET powder at 55 °C within half a day in a pH-stat bioreactor.

Suggested Citation

  • Hwaseok Hong & Dongwoo Ki & Hogyun Seo & Jiyoung Park & Jaewon Jang & Kyung-Jin Kim, 2023. "Discovery and rational engineering of PET hydrolase with both mesophilic and thermophilic PET hydrolase properties," Nature Communications, Nature, vol. 14(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-40233-w
    DOI: 10.1038/s41467-023-40233-w
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    References listed on IDEAS

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    1. Erika Erickson & Japheth E. Gado & Luisana Avilán & Felicia Bratti & Richard K. Brizendine & Paul A. Cox & Raj Gill & Rosie Graham & Dong-Jin Kim & Gerhard König & William E. Michener & Saroj Poudel &, 2022. "Sourcing thermotolerant poly(ethylene terephthalate) hydrolase scaffolds from natural diversity," Nature Communications, Nature, vol. 13(1), pages 1-15, December.
    2. Seongjoon Joo & In Jin Cho & Hogyun Seo & Hyeoncheol Francis Son & Hye-Young Sagong & Tae Joo Shin & So Young Choi & Sang Yup Lee & Kyung-Jin Kim, 2018. "Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation," Nature Communications, Nature, vol. 9(1), pages 1-12, December.
    3. Hongyuan Lu & Daniel J. Diaz & Natalie J. Czarnecki & Congzhi Zhu & Wantae Kim & Raghav Shroff & Daniel J. Acosta & Bradley R. Alexander & Hannah O. Cole & Yan Zhang & Nathaniel A. Lynd & Andrew D. El, 2022. "Machine learning-aided engineering of hydrolases for PET depolymerization," Nature, Nature, vol. 604(7907), pages 662-667, April.
    4. V. Tournier & C. M. Topham & A. Gilles & B. David & C. Folgoas & E. Moya-Leclair & E. Kamionka & M.-L. Desrousseaux & H. Texier & S. Gavalda & M. Cot & E. Guémard & M. Dalibey & J. Nomme & G. Cioci & , 2020. "An engineered PET depolymerase to break down and recycle plastic bottles," Nature, Nature, vol. 580(7802), pages 216-219, April.
    5. Xu Han & Weidong Liu & Jian-Wen Huang & Jiantao Ma & Yingying Zheng & Tzu-Ping Ko & Limin Xu & Ya-Shan Cheng & Chun-Chi Chen & Rey-Ting Guo, 2017. "Structural insight into catalytic mechanism of PET hydrolase," Nature Communications, Nature, vol. 8(1), pages 1-6, December.
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    Cited by:

    1. Elizabeth L. Bell & Gloria Rosetto & Morgan A. Ingraham & Kelsey J. Ramirez & Clarissa Lincoln & Ryan W. Clarke & Japheth E. Gado & Jacob L. Lilly & Katarzyna H. Kucharzyk & Erika Erickson & Gregg T. , 2024. "Natural diversity screening, assay development, and characterization of nylon-6 enzymatic depolymerization," Nature Communications, Nature, vol. 15(1), pages 1-17, December.
    2. Yinglu Cui & Yanchun Chen & Jinyuan Sun & Tong Zhu & Hua Pang & Chunli Li & Wen-Chao Geng & Bian Wu, 2024. "Computational redesign of a hydrolase for nearly complete PET depolymerization at industrially relevant high-solids loading," Nature Communications, Nature, vol. 15(1), pages 1-12, December.

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