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Characterization of a poly(butylene adipate-co-terephthalate)-hydrolyzing lipase from Pelosinus fermentans

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Abstract

Certain α/β hydrolases have the ability to hydrolyze synthetic polyesters. While their partial hydrolysis has a potential for surface functionalization, complete hydrolysis allows recycling of valuable building blocks. Although knowledge about biodegradation of these materials is important regarding their fate in the environment, it is currently limited to aerobic organisms. A lipase from the anaerobic groundwater organism Pelosinus fermentans DSM 17108 (PfL1) was cloned and expressed in Escherichia coli BL21-Gold(DE3) and purified from the cell extract. Biochemical characterization with small substrates showed thermoalkalophilic properties (T opt = 50 °C, pHopt = 7.5) and higher activity towards para-nitrophenyl octanoate (12.7 U mg−1) compared to longer and shorter chain lengths (C14 0.7 U mg−1 and C2 4.3 U mg−1, respectively). Crystallization and determination of the 3-D structure displayed the presence of a lid structure and a zinc ion surrounded by an extra domain. These properties classify the enzyme into the I.5 lipase family. PfL1 is able to hydrolyze poly(1,4-butylene adipate-co-terephthalate) (PBAT) polymeric substrates. The hydrolysis of PBAT showed the release of small building blocks as detected by liquid chromatography-mass spectrometry (LC-MS). Protein dynamics seem to be involved with lid opening for the hydrolysis of PBAT by PfL1.

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Acknowledgments

This project has received funding from the European Union’s Seventh Framework Programme for research, technological development, and demonstration under grant agreement no. 289253. This work has also been supported by the Federal Ministry of Science, Research and Economy (BMWFW), the federal Ministry of Traffic, Innovation and Technology (BMVIT), the Styrian Business Promotion Agency SFG, and the Standortagentur Tirol and ZIT-Technology Agency of the City of Vienna through the COMET-Funding Program managed by the Austrian Research Promotion Agency FFG. We highly appreciate the support of the beamline staff at the ESRF in Grenoble, France, during diffraction data collection. We acknowledge Stephanie Follonier at HES-SO Valais (Sion, Switzerland) for the help during the experiments with PHBV.

Conflict of interest

The authors declare no financial or commercial conflict of interest.

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Authors and Affiliations

  1. Institute of Environmental Biotechnology, University of Natural Resources and Life Science (BOKU), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria

    Antonino Biundo, Felice Quartinello, Doris Ribitsch & Georg M. Guebitz

  2. Austrian Centre for Industrial Biotechnology (ACIB), Petersgasse 14, 8010, Graz, Austria

    Altijana Hromic, Tea Pavkov-Keller & Karl Gruber

  3. Institute of Molecular Bioscience, University of Graz, Universitaetsstraße 15, 8010, Graz, Austria

    Altijana Hromic, Tea Pavkov-Keller & Karl Gruber

  4. Austrian Centre for Industrial Biotechnology (ACIB), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria

    Karolina Haernvall, Veronika Perz, Doris Ribitsch & Georg M. Guebitz

  5. Institute of Life Technologies, University of Applied Sciences Western Switzerland (HES-SO Valais/Wallis), Rue du Rawil 47, 1950, Sion, Switzerland

    Miriam S. Arrell & Manfred Zinn

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  1. Antonino Biundo
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Biundo, A., Hromic, A., Pavkov-Keller, T. et al. Characterization of a poly(butylene adipate-co-terephthalate)-hydrolyzing lipase from Pelosinus fermentans . Appl Microbiol Biotechnol 100, 1753–1764 (2016). https://doi.org/10.1007/s00253-015-7031-1

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