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

A polyethylene terephthalate (PET)-degrading bacterium identified as <i>Stenotrophomonas maltophilia</i> 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...

Full description

Bibliographic Details
Main Authors: Salah Ud Din, Kalsoom, Sadia Mehmood Satti, Salah Uddin, Smita V. Mankar, Esma Ceylan, Fariha Hasan, Samiullah Khan, Malik Badshah, Ali Osman Beldüz, Sabriye Çanakçi, Baozhong Zhang, Javier A. Linares-Pastén, Aamer Ali Shah
Format: Article
Language:English
Published: MDPI AG 2023-03-01
Series:Applied Sciences
Subjects:
Online Access:https://www.mdpi.com/2076-3417/13/6/3686
_version_ 1797613648011591680
author Salah Ud Din
Kalsoom
Sadia Mehmood Satti
Salah Uddin
Smita V. Mankar
Esma Ceylan
Fariha Hasan
Samiullah Khan
Malik Badshah
Ali Osman Beldüz
Sabriye Çanakçi
Baozhong Zhang
Javier A. Linares-Pastén
Aamer Ali Shah
author_facet Salah Ud Din
Kalsoom
Sadia Mehmood Satti
Salah Uddin
Smita V. Mankar
Esma Ceylan
Fariha Hasan
Samiullah Khan
Malik Badshah
Ali Osman Beldüz
Sabriye Çanakçi
Baozhong Zhang
Javier A. Linares-Pastén
Aamer Ali Shah
author_sort Salah Ud Din
collection DOAJ
description A polyethylene terephthalate (PET)-degrading bacterium identified as <i>Stenotrophomonas maltophilia</i> 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 (NiCl<sub>2</sub> and Na<sub>2</sub>SO<sub>4</sub>). The extracellular medium containing the cutinase-type enzyme showed a depolymerization yield of the PET powder comparable to that of <i>Idonella skaiensis Is</i>PETase and significantly higher than that of <i>Humicola insolens</i> thermostable HiCut (HiC) cutinase. This study suggests that <i>S. maltophilia</i> PRS8 is able to degrade PET at a mesophilic temperature and could be further explored for the sustainable management of plastic waste.
first_indexed 2024-03-11T06:58:48Z
format Article
id doaj.art-9cd9529b3cf440e5bfb3fc8df7020588
institution Directory Open Access Journal
issn 2076-3417
language English
last_indexed 2024-03-11T06:58:48Z
publishDate 2023-03-01
publisher MDPI AG
record_format Article
series Applied Sciences
spelling doaj.art-9cd9529b3cf440e5bfb3fc8df70205882023-11-17T09:25:22ZengMDPI AGApplied Sciences2076-34172023-03-01136368610.3390/app13063686The 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 TemperatureSalah Ud Din0Kalsoom1Sadia Mehmood Satti2Salah Uddin3Smita V. Mankar4Esma Ceylan5Fariha Hasan6Samiullah Khan7Malik Badshah8Ali Osman Beldüz9Sabriye Çanakçi10Baozhong Zhang11Javier A. Linares-Pastén12Aamer Ali Shah13Department of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, PakistanDepartment of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, PakistanDepartment of Microbiology, Kohsar University Murree, Murree 47150, PakistanDepartment of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, PakistanCentre for Analysis and Synthesis, Department of Chemistry, Faculty of Engineering (LTH), Lund University, SE-22100 Lund, SwedenDepartment of Biology, Karadeniz Technical University, Trabzon 61080, TurkeyDepartment of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, PakistanDepartment of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, PakistanDepartment of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, PakistanDepartment of Biology, Karadeniz Technical University, Trabzon 61080, TurkeyDepartment of Biology, Karadeniz Technical University, Trabzon 61080, TurkeyCentre for Analysis and Synthesis, Department of Chemistry, Faculty of Engineering (LTH), Lund University, SE-22100 Lund, SwedenDivision of Biotechnology, Department of Chemistry, Faculty of Engineering (LTH), Lund University, P.O. Box 124, SE-22100 Lund, SwedenDepartment of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, PakistanA polyethylene terephthalate (PET)-degrading bacterium identified as <i>Stenotrophomonas maltophilia</i> 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 (NiCl<sub>2</sub> and Na<sub>2</sub>SO<sub>4</sub>). The extracellular medium containing the cutinase-type enzyme showed a depolymerization yield of the PET powder comparable to that of <i>Idonella skaiensis Is</i>PETase and significantly higher than that of <i>Humicola insolens</i> thermostable HiCut (HiC) cutinase. This study suggests that <i>S. maltophilia</i> PRS8 is able to degrade PET at a mesophilic temperature and could be further explored for the sustainable management of plastic waste.https://www.mdpi.com/2076-3417/13/6/3686biodegradationPET<i>Stenotrophomonas maltophilia</i>cutinasePlackett–Burman designcentral composite design
spellingShingle Salah Ud Din
Kalsoom
Sadia Mehmood Satti
Salah Uddin
Smita V. Mankar
Esma Ceylan
Fariha Hasan
Samiullah Khan
Malik Badshah
Ali Osman Beldüz
Sabriye Çanakçi
Baozhong Zhang
Javier A. Linares-Pastén
Aamer Ali Shah
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
Applied Sciences
biodegradation
PET
<i>Stenotrophomonas maltophilia</i>
cutinase
Plackett–Burman design
central composite design
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
title_full 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
title_fullStr 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
title_full_unstemmed 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
title_short 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
title_sort 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
topic biodegradation
PET
<i>Stenotrophomonas maltophilia</i>
cutinase
Plackett–Burman design
central composite design
url https://www.mdpi.com/2076-3417/13/6/3686
work_keys_str_mv AT salahuddin thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT kalsoom thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT sadiamehmoodsatti thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT salahuddin thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT smitavmankar thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT esmaceylan thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT farihahasan thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT samiullahkhan thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT malikbadshah thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT aliosmanbelduz thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT sabriyecanakci thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT baozhongzhang thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT javieralinarespasten thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT aameralishah thepurificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT salahuddin purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT kalsoom purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT sadiamehmoodsatti purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT salahuddin purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT smitavmankar purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT esmaceylan purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT farihahasan purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT samiullahkhan purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT malikbadshah purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT aliosmanbelduz purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT sabriyecanakci purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT baozhongzhang purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT javieralinarespasten purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature
AT aameralishah purificationandcharacterizationofacutinaselikeenzymewithactivityonpolyethyleneterephthalatepetfromanewlyisolatedbacteriumistenotrophomonasmaltophiliaiprs8atamesophilictemperature