Novel efficient enzymatic synthesis of the key-reaction intermediate of PET depolymerization, mono(2-hydroxyethyl terephthalate) – MHET

Eugenio, Erika de Queiros; Campisano, Ivone Sampaio Pereira; Dias, Ayres G.; Castro, Aline Machado de; Coelho, Maria Alice Zarur; Langone, Marta A. P.

doi:10.1016/j.jbiotec.2022.08.019

Cited by 6 publications

(12 citation statements)

References 51 publications

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“…This suggests that, in addition to mutations around the active site, the pH and pD conditions may be used to tune the relative amounts of degradation products, which could be of interest for optimizing the enzymatic synthesis of MHET by cutinases. 30 In contrast to the PET powder assay, where pH 9.0 gave the highest activity, time-resolved NMR assays on PET films at different pD values showed that pD 6.5 (and not pD 9.0) resulted in maximum enzymatic activity (Figure 3). Ronkvist et al 99 have previously observed that FsC activity on PET varies little from pH 6.5 to 8.5, but it drops sharply at pH 9.0.…”

Section: ■ Results and Discussionmentioning

confidence: 87%

“…At pD 9.0, the decrease in MHET (and increase in TPA) concentration after 400 min appears to be slower than at the other pD values (Figure ). This suggests that, in addition to mutations around the active site, the pH and pD conditions may be used to tune the relative amounts of degradation products, which could be of interest for optimizing the enzymatic synthesis of MHET by cutinases …”

Section: Resultsmentioning

confidence: 99%

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Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi

et al. 2023

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In recent years, the drawbacks of plastics have become evident, with plastic pollution becoming a major environmental issue. There is an urgent need to find solutions to efficiently manage plastic waste by using novel recycling methods. Biocatalytic recycling of plastics by using enzyme-catalyzed hydrolysis is one such solution that has gained interest, in particular for recycling poly(ethylene terephthalate) (PET). To provide insights into PET hydrolysis by cutinases, we have here characterized the kinetics of a PET-hydrolyzing cutinase from Fusarium solani pisi (FsC) at different pH values, mapped the interaction between FsC and the PET analogue BHET by using NMR spectroscopy, and monitored product release directly and in real time by using time-resolved NMR experiments. We found that primarily aliphatic side chains around the active site participate in the interaction with BHET and that pH conditions and a mutation around the active site (L182A) can be used to tune the relative amounts of degradation products. Moreover, we propose that the low catalytic performance of FsC on PET is caused by poor substrate binding combined with slow MHET hydrolysis. Overall, our results provide insights into obstacles that preclude efficient PET hydrolysis by FsC and suggest future approaches for overcoming these obstacles and generating efficient PET-hydrolyzing enzymes.

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Section: ■ Results and Discussionmentioning

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi

et al. 2023

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show abstract

Section: Resultsmentioning

confidence: 99%

“…At pD 9.0, the decrease in MHET (and increase in TPA) concentration appears to be slower than at the other pD values. This suggests that the pH and pD conditions can be used to tune the relative amounts of degradation products, which may be of interest for optimization of enzymatic synthesis of MHET by cutinases 30 . and δMHET,H1 = 7.94 ppm (doublet) and δMHET,H2 = 8.12 ppm, where H1 corresponds to the "TPA" side and H2 corresponds to the "ethylene glycol" side of the aromatic ring of MHET.…”

Section: Hydrolytic Activity On Pet Films Monitored By Time-resolved Nmrmentioning

confidence: 99%

“…At pD 9.0, the decrease in MHET (and increase in TPA) concentration after 400 minutes appears to be slower than at the other pD values (Figure 3). This suggests that, in addition to mutations around the active site, the pH and pD conditions may be used to tune the relative amounts of degradation products, which could be of interest for optimizing enzymatic synthesis of MHET by cutinases 30 . In contrast with the PET powder assay where pH 9.0 gave highest activity, time-resolved NMR assays on PET films at different pD values showed that pD 6.5 (and not pD 9.0) resulted in maximum enzymatic activity (Figure 3).…”

Section: Hydrolytic Activity On Pet Films Monitored By Time-resolved Nmrmentioning

confidence: 99%

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Biochemical characterization and NMR study of a PET-hydrolyzing cutinase fromFusarium solani pisi

Hellesnes

Vijayaraj

Fojan

et al. 2022

Preprint

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In recent years, the drawbacks of plastics have become evident, with plastic pollution becoming a major environmental issue. There is an urgent need to find solutions to efficiently handle plastic waste by using novel recycling methods. Biocatalytic recycling of plastics by using enzyme-catalyzed hydrolysis is one such solution that has gained interest, in particular for recycling polyethylene terephthalate (PET). To provide insights on PET hydrolysis by cutinases, we have here characterized the kinetics of a PET-hydrolyzing cutinase fromFusarium solani pisi(FsC) at different pH values, mapped the interaction between FsC and the PET analog BHET by using NMR spectroscopy, and monitored product release directly and in real time by using time-resolved NMR experiments. We found that primarily aliphatic side chains around the active site participate in the interaction with BHET, and that pH conditions can be used to tune the relative amounts of degradation products. Moreover, we propose that the low catalytic performance of FsC on PET is caused by poor substrate binding combined with product inhibition by MHET. Overall, our results provide insights on obstacles that preclude efficient PET hydrolysis by FsC and suggest future approaches for overcoming these obstacles and generating efficient PET-hydrolyzing enzymes.

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Unlocking a Sustainable Future for Plastics: A Chemical‐Enzymatic Pathway for Efficient Conversion of Mixed Waste to MHET and Energy‐Saving PET Recycling

Li,

Wu,

Cui

et al. 2024

ChemSusChem

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The heterogeneous monomers obtained from plastic waste degradation are unfavorable for PET recondensation and high‐value derivative synthesis. Herein, we developed an efficient chemical‐enzymatic approach to convert mixed plastic wastes into homogeneous mono‐2‐hydroxyethyl terephthalate (MHET) without downstream purification, benefiting from three discovered BHETases (KbEst, KbHyd, and BrevEst) in nature. Towards the mixed plastic waste, integrating the chemical K2CO3‐driven glycolysis process with the BHETase depolymerization technique resulted in an MHET yield of up to 98.26% in 40 h. Remarkably, BrevEst accomplished the highest BHET hydrolysis (~87% efficiency in 12 h) for yielding analytical‐grade MHET compared to seven state‐of‐the‐art PET hydrolases (18%‐40%). In an investigation combining quantum theoretical computations and experimental validations, we established a MHET‐initiated PET repolymerization pathway. This shortcut approach with MHET promises to strengthen the valorization of mixed plastics, offering a substantially more efficient and energy‐saving route for PET recycling.

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Novel efficient enzymatic synthesis of the key-reaction intermediate of PET depolymerization, mono(2-hydroxyethyl terephthalate) – MHET

Cited by 6 publications

References 51 publications

Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi

Biochemical Characterization and NMR Study of a PET-Hydrolyzing Cutinase from Fusarium solani pisi

Biochemical characterization and NMR study of a PET-hydrolyzing cutinase fromFusarium solani pisi

Unlocking a Sustainable Future for Plastics: A Chemical‐Enzymatic Pathway for Efficient Conversion of Mixed Waste to MHET and Energy‐Saving PET Recycling

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