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...
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2023-03-01
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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. |
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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 |
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