The Purification and Characterization of a Cutinase-like Enzyme with Activity on Polyethylene Terephthalate (PET) from a Newly Isolated Bacterium Stenotrophomonas maltophilia PRS8 at a Mesophilic Temperature
(2023) In Applied Sciences (Switzerland) 13(6).- Abstract
- A polyethylene terephthalate (PET)-degrading bacterium identified as Stenotrophomonas maltophilia
PRS8 was isolated from the soil of a landfill. The degradation of the
PET bottle flakes and the PET prepared as a powder were assessed using
live cells, an extracellular medium, or a purified cutinase-like enzyme.
These treated polymers were analyzed using Fourier transform infrared
spectroscopy (FTIR) and scanning electron microscopy (SEM). The
depolymerization products, identified using HPLC and LC-MS, were
terephthalic acid (TPA), mono(2-hydroxyethyl)-TPA (MHET), and
bis(2-hydroxyethyl)-TPA (BHET). Several physicochemical factors were
optimized for a better cutinase-like enzyme production by... (More) - A polyethylene terephthalate (PET)-degrading bacterium identified as Stenotrophomonas maltophilia
PRS8 was isolated from the soil of a landfill. The degradation of the
PET bottle flakes and the PET prepared as a powder were assessed using
live cells, an extracellular medium, or a purified cutinase-like enzyme.
These treated polymers were analyzed using Fourier transform infrared
spectroscopy (FTIR) and scanning electron microscopy (SEM). The
depolymerization products, identified using HPLC and LC-MS, were
terephthalic acid (TPA), mono(2-hydroxyethyl)-TPA (MHET), and
bis(2-hydroxyethyl)-TPA (BHET). Several physicochemical factors were
optimized for a better cutinase-like enzyme production by using unique
single-factor and multi-factor statistical models (the Plackett–Burman
design and the central composite design software). The enzyme was
purified for homogeneity through column chromatography using Sephadex
G-100 resin. The molecular weight of the enzyme was approximately 58
kDa. The specific activity on para nitrophenyl butyrate was estimated at
450.58 U/mg, with a purification of 6.39 times and a yield of 48.64%.
The enzyme was stable at various temperatures (30–40 °C) and pH levels
(8.0–10.0). The enzyme activity was significantly improved by the
surfactants (Triton X-100 and Tween-40), organic solvent (formaldehyde),
and metals (NiCl2 and Na2SO4). The
extracellular medium containing the cutinase-type enzyme showed a
depolymerization yield of the PET powder comparable to that of Idonella skaiensis IsPETase and significantly higher than that of Humicola insolens thermostable HiCut (HiC) cutinase. This study suggests that S. maltophilia
PRS8 is able to degrade PET at a mesophilic temperature and could be
further explored for the sustainable management of plastic waste. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/c8fc65d7-5b8c-4bbf-b961-5dfce7ae0333
- author
- organization
- publishing date
- 2023-03-14
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- biodegradation, PET, Stenotrophomonas maltophilia, cutinase, Plackett–Burman design, central composite design
- in
- Applied Sciences (Switzerland)
- volume
- 13
- issue
- 6
- article number
- 3686
- pages
- 22 pages
- publisher
- MDPI AG
- external identifiers
-
- scopus:85151533815
- ISSN
- 2076-3417
- DOI
- 10.3390/app13063686
- language
- English
- LU publication?
- yes
- id
- c8fc65d7-5b8c-4bbf-b961-5dfce7ae0333
- date added to LUP
- 2023-03-14 13:55:18
- date last changed
- 2023-05-23 11:27:01
@article{c8fc65d7-5b8c-4bbf-b961-5dfce7ae0333, abstract = {{A polyethylene terephthalate (PET)-degrading bacterium identified as Stenotrophomonas maltophilia<br> PRS8 was isolated from the soil of a landfill. The degradation of the <br> PET bottle flakes and the PET prepared as a powder were assessed using <br> live cells, an extracellular medium, or a purified cutinase-like enzyme.<br> These treated polymers were analyzed using Fourier transform infrared <br> spectroscopy (FTIR) and scanning electron microscopy (SEM). The <br> depolymerization products, identified using HPLC and LC-MS, were <br> terephthalic acid (TPA), mono(2-hydroxyethyl)-TPA (MHET), and <br> bis(2-hydroxyethyl)-TPA (BHET). Several physicochemical factors were <br> optimized for a better cutinase-like enzyme production by using unique <br> single-factor and multi-factor statistical models (the Plackett–Burman <br> design and the central composite design software). The enzyme was <br> purified for homogeneity through column chromatography using Sephadex <br> G-100 resin. The molecular weight of the enzyme was approximately 58 <br> kDa. The specific activity on para nitrophenyl butyrate was estimated at<br> 450.58 U/mg, with a purification of 6.39 times and a yield of 48.64%. <br> The enzyme was stable at various temperatures (30–40 °C) and pH levels <br> (8.0–10.0). The enzyme activity was significantly improved by the <br> surfactants (Triton X-100 and Tween-40), organic solvent (formaldehyde),<br> and metals (NiCl<sub>2</sub> and Na<sub>2</sub>SO<sub>4</sub>). The <br> extracellular medium containing the cutinase-type enzyme showed a <br> depolymerization yield of the PET powder comparable to that of Idonella skaiensis IsPETase and significantly higher than that of Humicola insolens thermostable HiCut (HiC) cutinase. This study suggests that S. maltophilia<br> PRS8 is able to degrade PET at a mesophilic temperature and could be <br> further explored for the sustainable management of plastic waste.}}, author = {{Din, Salah Ud and Kalsoom and Satti, Sadia Mehmood and Uddin, Salah and Mankar, Smita V. and Ceylan, Esma and Hasan, Fariha and Khan, Samiullah and Badshah, Malik and Beldüz, Ali Osman and Çanakçi, Sabriye and Zhang, Baozhong and Linares-Pastén, Javier A. and Shah, Aamer Ali}}, issn = {{2076-3417}}, keywords = {{biodegradation; PET; Stenotrophomonas maltophilia; cutinase; Plackett–Burman design; central composite design}}, language = {{eng}}, month = {{03}}, number = {{6}}, publisher = {{MDPI AG}}, series = {{Applied Sciences (Switzerland)}}, title = {{The Purification and Characterization of a Cutinase-like Enzyme with Activity on Polyethylene Terephthalate (PET) from a Newly Isolated Bacterium <i>Stenotrophomonas maltophilia</i> PRS8 at a Mesophilic Temperature}}, url = {{http://dx.doi.org/10.3390/app13063686}}, doi = {{10.3390/app13063686}}, volume = {{13}}, year = {{2023}}, }