Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1
Аутори
Lončar, NikolaDrašković, Natalija
Božić, Nataša
Romero, Elvira
Simić, Stefan
Opsenica, Igor
Vujčić, Zoran
Fraaije, Marco W.
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
The consumption of dyes is increasing worldwide in line with the increase of population
and demand for clothes and other colored products. However, the e ciency of dyeing processes
is still poor and results in large amounts of colored e uents. It is desired to develop a portfolio
of enzymes which can be used for the treatment of colored wastewaters. Herein, we used genome
sequence information to discover a dye-decolorizing peroxidase (DyP) from Pseudomonas fluorescens Pf-01. Two genes putatively encoding for DyPs were identified in the respective genome and cloned for expression in Escherichia coli, of which one (PfDyP B2) could be overexpressed as a soluble protein. PfDyP B2 shows some typical features known for DyPs which includes the ability to convert dyes at the expense of hydrogen peroxide. Interestingly, t-butyl hydroperoxide could be used as an alternative substrate to hydrogen peroxide. Immobilization of PfDyP B2 in calcium-alginate beads resulted in a significant increase... in stability: PfDyP B2 retains 80% of its initial activity after 2 h incubation at 50 °C, while the soluble enzyme is inactivated within minutes. PfDyP B2 was also tested with aniline and ethyl diazoacetate as substrates. Based on GC-MS analyses, 30% conversion of the starting material was achieved after 65 h at 30 °C. Importantly, this is the first report of a DyP-catalyzed insertion of a carbene into an N-H bond.
Кључне речи:
DyP peroxidase / oxidoreductase / reactive dye / decolorizationИзвор:
Catalysts, 2019, 9, 463-Издавач:
- MDPI
Финансирање / пројекти:
- Производња, изоловање и карактеризација ензима и малих молекула и њихова примена у растворном и имобилизованом облику у биотехнологији хране, биогоривима и заштитити животне средине (RS-MESTD-Basic Research (BR or ON)-172048)
DOI: 10.3390/catal9050463
ISSN: 2073-4344
WoS: 000472678500072
Scopus: 2-s2.0-85066730056
Институција/група
IHTMTY - JOUR AU - Lončar, Nikola AU - Drašković, Natalija AU - Božić, Nataša AU - Romero, Elvira AU - Simić, Stefan AU - Opsenica, Igor AU - Vujčić, Zoran AU - Fraaije, Marco W. PY - 2019 UR - https://cer.ihtm.bg.ac.rs/handle/123456789/3021 AB - The consumption of dyes is increasing worldwide in line with the increase of population and demand for clothes and other colored products. However, the e ciency of dyeing processes is still poor and results in large amounts of colored e uents. It is desired to develop a portfolio of enzymes which can be used for the treatment of colored wastewaters. Herein, we used genome sequence information to discover a dye-decolorizing peroxidase (DyP) from Pseudomonas fluorescens Pf-01. Two genes putatively encoding for DyPs were identified in the respective genome and cloned for expression in Escherichia coli, of which one (PfDyP B2) could be overexpressed as a soluble protein. PfDyP B2 shows some typical features known for DyPs which includes the ability to convert dyes at the expense of hydrogen peroxide. Interestingly, t-butyl hydroperoxide could be used as an alternative substrate to hydrogen peroxide. Immobilization of PfDyP B2 in calcium-alginate beads resulted in a significant increase in stability: PfDyP B2 retains 80% of its initial activity after 2 h incubation at 50 °C, while the soluble enzyme is inactivated within minutes. PfDyP B2 was also tested with aniline and ethyl diazoacetate as substrates. Based on GC-MS analyses, 30% conversion of the starting material was achieved after 65 h at 30 °C. Importantly, this is the first report of a DyP-catalyzed insertion of a carbene into an N-H bond. PB - MDPI T2 - Catalysts T1 - Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1 VL - 9 SP - 463 DO - 10.3390/catal9050463 ER -
@article{ author = "Lončar, Nikola and Drašković, Natalija and Božić, Nataša and Romero, Elvira and Simić, Stefan and Opsenica, Igor and Vujčić, Zoran and Fraaije, Marco W.", year = "2019", abstract = "The consumption of dyes is increasing worldwide in line with the increase of population and demand for clothes and other colored products. However, the e ciency of dyeing processes is still poor and results in large amounts of colored e uents. It is desired to develop a portfolio of enzymes which can be used for the treatment of colored wastewaters. Herein, we used genome sequence information to discover a dye-decolorizing peroxidase (DyP) from Pseudomonas fluorescens Pf-01. Two genes putatively encoding for DyPs were identified in the respective genome and cloned for expression in Escherichia coli, of which one (PfDyP B2) could be overexpressed as a soluble protein. PfDyP B2 shows some typical features known for DyPs which includes the ability to convert dyes at the expense of hydrogen peroxide. Interestingly, t-butyl hydroperoxide could be used as an alternative substrate to hydrogen peroxide. Immobilization of PfDyP B2 in calcium-alginate beads resulted in a significant increase in stability: PfDyP B2 retains 80% of its initial activity after 2 h incubation at 50 °C, while the soluble enzyme is inactivated within minutes. PfDyP B2 was also tested with aniline and ethyl diazoacetate as substrates. Based on GC-MS analyses, 30% conversion of the starting material was achieved after 65 h at 30 °C. Importantly, this is the first report of a DyP-catalyzed insertion of a carbene into an N-H bond.", publisher = "MDPI", journal = "Catalysts", title = "Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1", volume = "9", pages = "463", doi = "10.3390/catal9050463" }
Lončar, N., Drašković, N., Božić, N., Romero, E., Simić, S., Opsenica, I., Vujčić, Z.,& Fraaije, M. W.. (2019). Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1. in Catalysts MDPI., 9, 463. https://doi.org/10.3390/catal9050463
Lončar N, Drašković N, Božić N, Romero E, Simić S, Opsenica I, Vujčić Z, Fraaije MW. Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1. in Catalysts. 2019;9:463. doi:10.3390/catal9050463 .
Lončar, Nikola, Drašković, Natalija, Božić, Nataša, Romero, Elvira, Simić, Stefan, Opsenica, Igor, Vujčić, Zoran, Fraaije, Marco W., "Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1" in Catalysts, 9 (2019):463, https://doi.org/10.3390/catal9050463 . .