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Full text release has been delayed at the author's request until July 18, 2029

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Scale-Up Production and Stability Assessment of the Petase Enzyme

MacFarlane, Christopher Edward
http://orcid.org/0009-0001-2226-8505
http://rave.ohiolink.edu/etdc/view?acc_num=miami1721332327239787

Abstract Details

2024, Master of Science, Miami University, Chemical, Paper and Biomedical Engineering.
In 2021, over 390 million metric tons of plastic were produced, with a large quantity being PET, which is commonly used for single use plastics such as disposable water bottles. PET is engineered for durability, resulting in a slow natural breakdown. A recently discovered enzyme, PETase, speeds up PET depolymerization; however, it’s turnover rate, stability, and production quantity are too low to be used in practical applications. The goal of this work is to address these problems. We assessed the production of PETase using different medias, carbon sources, and induction methods. For expression, PETase activity was measured in clarified lysates by quantifying catalysis on p-nitrophenyl acetate. Using common laboratory medias in a bioreactor, PETase expression levels were increased 1.5-fold. Using a complex media, with glycerol as a carbon source, a further 3-fold increase in expression occurred with increased batch time, reaching greater than 100 mg per L. Activity of PETase seems to limit its ability to catalyze reactions on PET substrates in long reactions. Experiments were preformed to understand and improve the PETase degradation of PET under industrially-relevant conditions. We expect these advances in expression, and characterization of PETase will help improve the application of this enzyme to recycle plastics.
Jason Berberich (Advisor)
Jason Boock (Advisor)
Keith Hohn (Committee Member)
49 p.
Cellular Biology; Civil Engineering; Sustainability
PETase; enzyme; plastic; recycling; plastic recycling; scale-up; stability; bioreactor; proteins; bacteria; E. coli; protein expression; PET; polyethylene terephthalate;

Recommended Citations

Citations

  • MacFarlane, C. E. (2024). Scale-Up Production and Stability Assessment of the Petase Enzyme [Master's thesis, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=miami1721332327239787

    APA Style (7th edition)

  • MacFarlane, Christopher. Scale-Up Production and Stability Assessment of the Petase Enzyme. 2024. Miami University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=miami1721332327239787.

    MLA Style (8th edition)

  • MacFarlane, Christopher. "Scale-Up Production and Stability Assessment of the Petase Enzyme." Master's thesis, Miami University, 2024. http://rave.ohiolink.edu/etdc/view?acc_num=miami1721332327239787

    Chicago Manual of Style (17th edition)

miami1721332327239787
© 2024, some rights reserved.Creative Commons License Scale-Up Production and Stability Assessment of the Petase Enzyme by Christopher Edward MacFarlane is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by Miami University and OhioLINK.