Saturation mutagenesis to improve the degradation of azo dyes by versatile peroxidase and application in form of VP-coated yeast cell walls
Само за регистроване кориснике
2020
Аутори
Ilić Đurđić, KarlaOstafe, Raluca
Đurđević Đelmaš, Aleksandra
Popović, Nikolina
Schillberg, Stefan
Fischer, Rainer
Prodanović, Radivoje
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Azo dyes are toxic and carcinogenic synthetic pigments that accumulate as pollutants in aquatic bodies near textile factories. The pigments are structurally diverse, and bioremediation is mostly limited to single dye compounds or related groups. Versatile peroxidase (VP) from Pleurotus eryngii is a heme-containing peroxidase with a broad substrate spectrum that can break down many structurally distinct pollutants, including azo dyes. The utilization of this enzyme could be facilitated by engineering to modify its catalytic activity and substrate range. We used saturation mutagenesis to alter two amino acids in the catalytic tryptophan environment of VP (V160 and A260). Library screening with three azo dyes revealed that these two positions had a significant influence on substrate specificity. We were able to isolate and sequence VP variants with up to 16-fold higher catalytic efficiency for different azo dyes. The same approach could be used to select for VP variants that catalyze the ...degradation of many other types of pollutants. To allow multiple cycles of dye degradation, we immobilized VP on the surface of yeast cells and used washed cell wall fragments after lysis. VP embedded in the cell wall retained ∼70 % of its initial activity after 10 cycles of dye degradation each lasting 12 h, making this platform ideal for the bioremediation of environments contaminated with azo dyes.
Кључне речи:
Cell wall fragments / Dye degradation / Pleurotus eryngii / Protein engineering / Yeast surface displayИзвор:
Enzyme and Microbial Technology, 2020, 136, e109509-Издавач:
- Elsevier
Финансирање / пројекти:
- Испитивања односа структура-функција у ћелијском зиду биљака и измене структуре зида ензимским инжењерингом (RS-173017)
- Алергени, антитела, ензими и мали физиолошки значајни молекули: дизајн, структура, функција и значај (RS-172049)
- Развој нових инкапсулационих и ензимских технологија за производњу биокатализатора и биолошки активних компонената хране у циљу повећања њене конкурентности, квалитета и безбедности (RS-46010)
Напомена:
- Peer-reviewed manuscript: http://cherry.chem.bg.ac.rs/handle/123456789/3835
- Supplementary material: http://cherry.chem.bg.ac.rs/handle/123456789/3836
DOI: 10.1016/j.enzmictec.2020.109509
ISSN: 0141-0229
WoS: 000528248400012
Scopus: 2-s2.0-85078512492
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Ilić Đurđić, Karla AU - Ostafe, Raluca AU - Đurđević Đelmaš, Aleksandra AU - Popović, Nikolina AU - Schillberg, Stefan AU - Fischer, Rainer AU - Prodanović, Radivoje PY - 2020 UR - https://cherry.chem.bg.ac.rs/handle/123456789/3834 AB - Azo dyes are toxic and carcinogenic synthetic pigments that accumulate as pollutants in aquatic bodies near textile factories. The pigments are structurally diverse, and bioremediation is mostly limited to single dye compounds or related groups. Versatile peroxidase (VP) from Pleurotus eryngii is a heme-containing peroxidase with a broad substrate spectrum that can break down many structurally distinct pollutants, including azo dyes. The utilization of this enzyme could be facilitated by engineering to modify its catalytic activity and substrate range. We used saturation mutagenesis to alter two amino acids in the catalytic tryptophan environment of VP (V160 and A260). Library screening with three azo dyes revealed that these two positions had a significant influence on substrate specificity. We were able to isolate and sequence VP variants with up to 16-fold higher catalytic efficiency for different azo dyes. The same approach could be used to select for VP variants that catalyze the degradation of many other types of pollutants. To allow multiple cycles of dye degradation, we immobilized VP on the surface of yeast cells and used washed cell wall fragments after lysis. VP embedded in the cell wall retained ∼70 % of its initial activity after 10 cycles of dye degradation each lasting 12 h, making this platform ideal for the bioremediation of environments contaminated with azo dyes. PB - Elsevier T2 - Enzyme and Microbial Technology T1 - Saturation mutagenesis to improve the degradation of azo dyes by versatile peroxidase and application in form of VP-coated yeast cell walls VL - 136 SP - e109509 DO - 10.1016/j.enzmictec.2020.109509 ER -
@article{ author = "Ilić Đurđić, Karla and Ostafe, Raluca and Đurđević Đelmaš, Aleksandra and Popović, Nikolina and Schillberg, Stefan and Fischer, Rainer and Prodanović, Radivoje", year = "2020", abstract = "Azo dyes are toxic and carcinogenic synthetic pigments that accumulate as pollutants in aquatic bodies near textile factories. The pigments are structurally diverse, and bioremediation is mostly limited to single dye compounds or related groups. Versatile peroxidase (VP) from Pleurotus eryngii is a heme-containing peroxidase with a broad substrate spectrum that can break down many structurally distinct pollutants, including azo dyes. The utilization of this enzyme could be facilitated by engineering to modify its catalytic activity and substrate range. We used saturation mutagenesis to alter two amino acids in the catalytic tryptophan environment of VP (V160 and A260). Library screening with three azo dyes revealed that these two positions had a significant influence on substrate specificity. We were able to isolate and sequence VP variants with up to 16-fold higher catalytic efficiency for different azo dyes. The same approach could be used to select for VP variants that catalyze the degradation of many other types of pollutants. To allow multiple cycles of dye degradation, we immobilized VP on the surface of yeast cells and used washed cell wall fragments after lysis. VP embedded in the cell wall retained ∼70 % of its initial activity after 10 cycles of dye degradation each lasting 12 h, making this platform ideal for the bioremediation of environments contaminated with azo dyes.", publisher = "Elsevier", journal = "Enzyme and Microbial Technology", title = "Saturation mutagenesis to improve the degradation of azo dyes by versatile peroxidase and application in form of VP-coated yeast cell walls", volume = "136", pages = "e109509", doi = "10.1016/j.enzmictec.2020.109509" }
Ilić Đurđić, K., Ostafe, R., Đurđević Đelmaš, A., Popović, N., Schillberg, S., Fischer, R.,& Prodanović, R.. (2020). Saturation mutagenesis to improve the degradation of azo dyes by versatile peroxidase and application in form of VP-coated yeast cell walls. in Enzyme and Microbial Technology Elsevier., 136, e109509. https://doi.org/10.1016/j.enzmictec.2020.109509
Ilić Đurđić K, Ostafe R, Đurđević Đelmaš A, Popović N, Schillberg S, Fischer R, Prodanović R. Saturation mutagenesis to improve the degradation of azo dyes by versatile peroxidase and application in form of VP-coated yeast cell walls. in Enzyme and Microbial Technology. 2020;136:e109509. doi:10.1016/j.enzmictec.2020.109509 .
Ilić Đurđić, Karla, Ostafe, Raluca, Đurđević Đelmaš, Aleksandra, Popović, Nikolina, Schillberg, Stefan, Fischer, Rainer, Prodanović, Radivoje, "Saturation mutagenesis to improve the degradation of azo dyes by versatile peroxidase and application in form of VP-coated yeast cell walls" in Enzyme and Microbial Technology, 136 (2020):e109509, https://doi.org/10.1016/j.enzmictec.2020.109509 . .