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Kinetics and mechanism of heterogeneous hydrolysis of poly[(R)-3-hydroxybutyrate] film by PHA depolymerases
Abstract
The kinetics and mechanism of enzymatic degradation on the surface of poly[(R)-3-hydroxybutyrate] (P[(R)-3HB]) film have been studied using three types of extracellular poly(hydroxyalkanoate) (PHA) depolymerases from Alcaligenes faecalis, Pseudomonas picketti and Comamonas testosteroni. The monomer and dimer of 3-hydroxybutyric acid were produced during the course of the enzymatic degradation of P[(R)-3HB] film, and the rate of production was determined by monitoring the increase in absorbance at 210 nm on a spectrophotometer. The rate of enzymatic degradation increased to a maximum value with the concentration of PHA depolymerase, followed by a gradual decrease. The kinetic data were accounted for in terms of a heterogeneous enzymatic reaction, involving enzymatic degradation on the surface of P[(R)-3HB] film via two steps of adsorption and hydrolysis by a PHA depolymerase with binding and catalytic domains. The kinetic results suggest that the properties of the catalytic domains are very similar among the three PHA depolymerases, but that those of the binding domains are strongly dependent on the type of depolymerase.
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“Because of this mechanism of action, adding a higher amount of enzyme increases the efficiency of the degradation up to an inhibitory concentration that is detrimental to the degradation rates, owing to saturation of the substrate that prevents the catalytic site from interacting with the polymer.497”
From Catalysis as an Enabling Science for Sustainable Polymers (2018) · cites this paper as [497]
In the citing paper
“Depending on the provenance of the PHB depolymerase, either monomer and dimer of 3-hydroxybutyric acid 497 or oligomers can be obtained.[498][499][500]”
From Catalysis as an Enabling Science for Sustainable Polymers (2018) · cites this paper as [497]
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Metadata
- Authors
- Mukai, K., Yamada, K., & Doi, Y.
- Affiliations
- RIKEN (JP)
- Corresponding author
- Doi, Yoshiharu
- Venue
- International Journal of Biological Macromolecules
- Publisher
- Elsevier BV
- Published
- 1993-12
- Volume
- 15
- Issue
- 6
- Pages
- 361-366
- ISSN
- 0141-8130 (ISSN-L)
- OpenAlex
- W2019749929
- Cited by
- 143
- References
- 27
- Citations by year
- 2026: 1 · 2025: 5 · 2024: 8 · 2023: 8 · 2022: 4 · 2021: 7 · 2020: 5 · 2019: 2 · 2018: 3 · 2017: 7 · 2016: 2 · 2015: 2 · 2014: 3 · 2013: 2 · 2012: 3
- FWCI
- 4.18
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- International Journal of Biological Macromolecules (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Materials Science › Biomaterials › biodegradable polymer synthesis and properties
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
- Physical Sciences › Environmental Science › Pollution › Microbial bioremediation and biosurfactants
- Keywords
- poly[(R)-3-hydroxybutyrate], PHA depolymerase, enzyme kinetics, enzyme mechanism, Alcaligenes faecalis, Comamonas testosteroni, beta-hydroxybutyrate, enzyme adsorption, binding domain, catalytic domain, biodegradable plastics
- MeSH terms
- Polyhydroxybutyrates, Alcaligenes, Carboxylic Ester Hydrolases, Hydrolysis, Hydroxybutyrates, Kinetics, Polyesters, Pseudomonas, Solubility
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