Semi‑rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide
Само за регистроване кориснике
2020
Аутори
Balaž, Ana MarijaStevanović, Jelena
Ostafe, Raluca
Blažić, Marija
Ilić Đurđić, Karla
Fischer, Rainer
Prodanović, Radivoje
Чланак у часопису (Објављена верзија)
,
Springer Nature Switzerland AG 2019
Метаподаци
Приказ свих података о документуАпстракт
Cellobiose dehydrogenase (CDH, EC 1.1.99.18) from white rot fungi Phanerochaete chrysosporium can be used for constructing biosensors and biofuel cells, for bleaching cotton in textile industry, and recently, the enzyme has found an important application in biomedicine as an antimicrobial and antibiofilm agent. Stability and activity of the wild-type (wt) CDH and mutants at methionine residues in the presence of hydrogen peroxide were investigated. Saturation mutagenesis libraries were made at the only methionine in heme domain M65 and two methionines M685 and M738 in the flavin domain that were closest to the active site. After screening the libraries, three mutants with increased activity and stability in the presence of peroxide were found, M65F with 70% of residual activity after 6 h of incubation in 0.3 M hydrogen peroxide, M738S with 80% of residual activity and M685Y with over 90% of residual activity compared to wild-type CDH that retained 40% of original activity. Combined mut...ants showed no activity. The most stable mutant M685Y with 5.8 times increased half-life in the presence of peroxide showed also 2.5 times increased kcat for lactose compared to wtCDH and could be good candidate for applications in biofuel cells and biocatalysis for lactobionic acid production.
Кључне речи:
Protein engineering / Library / Hydrogen peroxide / Methionine / YeastИзвор:
Molecular Diversity, 2020, 24, 593-601Издавач:
- Springer International Publishing
Финансирање / пројекти:
- Развој нових инкапсулационих и ензимских технологија за производњу биокатализатора и биолошки активних компонената хране у циљу повећања њене конкурентности, квалитета и безбедности (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-46010)
- Алергени, антитела, ензими и мали физиолошки значајни молекули: дизајн, структура, функција и значај (RS-MESTD-Basic Research (BR or ON)-172049)
- Испитивања односа структура-функција у ћелијском зиду биљака и измене структуре зида ензимским инжењерингом (RS-MESTD-Basic Research (BR or ON)-173017)
DOI: 10.1007/s11030-019-09965-0
ISSN: 1573-501; 1381-1991
PubMed: 31154590
WoS: 000547921900001
Scopus: 2-s2.0-85066612562
Институција/група
IHTMTY - JOUR AU - Balaž, Ana Marija AU - Stevanović, Jelena AU - Ostafe, Raluca AU - Blažić, Marija AU - Ilić Đurđić, Karla AU - Fischer, Rainer AU - Prodanović, Radivoje PY - 2020 UR - https://cer.ihtm.bg.ac.rs/handle/123456789/3128 AB - Cellobiose dehydrogenase (CDH, EC 1.1.99.18) from white rot fungi Phanerochaete chrysosporium can be used for constructing biosensors and biofuel cells, for bleaching cotton in textile industry, and recently, the enzyme has found an important application in biomedicine as an antimicrobial and antibiofilm agent. Stability and activity of the wild-type (wt) CDH and mutants at methionine residues in the presence of hydrogen peroxide were investigated. Saturation mutagenesis libraries were made at the only methionine in heme domain M65 and two methionines M685 and M738 in the flavin domain that were closest to the active site. After screening the libraries, three mutants with increased activity and stability in the presence of peroxide were found, M65F with 70% of residual activity after 6 h of incubation in 0.3 M hydrogen peroxide, M738S with 80% of residual activity and M685Y with over 90% of residual activity compared to wild-type CDH that retained 40% of original activity. Combined mutants showed no activity. The most stable mutant M685Y with 5.8 times increased half-life in the presence of peroxide showed also 2.5 times increased kcat for lactose compared to wtCDH and could be good candidate for applications in biofuel cells and biocatalysis for lactobionic acid production. PB - Springer International Publishing T2 - Molecular Diversity T1 - Semi‑rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide VL - 24 SP - 593 EP - 601 DO - 10.1007/s11030-019-09965-0 ER -
@article{ author = "Balaž, Ana Marija and Stevanović, Jelena and Ostafe, Raluca and Blažić, Marija and Ilić Đurđić, Karla and Fischer, Rainer and Prodanović, Radivoje", year = "2020", abstract = "Cellobiose dehydrogenase (CDH, EC 1.1.99.18) from white rot fungi Phanerochaete chrysosporium can be used for constructing biosensors and biofuel cells, for bleaching cotton in textile industry, and recently, the enzyme has found an important application in biomedicine as an antimicrobial and antibiofilm agent. Stability and activity of the wild-type (wt) CDH and mutants at methionine residues in the presence of hydrogen peroxide were investigated. Saturation mutagenesis libraries were made at the only methionine in heme domain M65 and two methionines M685 and M738 in the flavin domain that were closest to the active site. After screening the libraries, three mutants with increased activity and stability in the presence of peroxide were found, M65F with 70% of residual activity after 6 h of incubation in 0.3 M hydrogen peroxide, M738S with 80% of residual activity and M685Y with over 90% of residual activity compared to wild-type CDH that retained 40% of original activity. Combined mutants showed no activity. The most stable mutant M685Y with 5.8 times increased half-life in the presence of peroxide showed also 2.5 times increased kcat for lactose compared to wtCDH and could be good candidate for applications in biofuel cells and biocatalysis for lactobionic acid production.", publisher = "Springer International Publishing", journal = "Molecular Diversity", title = "Semi‑rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide", volume = "24", pages = "593-601", doi = "10.1007/s11030-019-09965-0" }
Balaž, A. M., Stevanović, J., Ostafe, R., Blažić, M., Ilić Đurđić, K., Fischer, R.,& Prodanović, R.. (2020). Semi‑rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide. in Molecular Diversity Springer International Publishing., 24, 593-601. https://doi.org/10.1007/s11030-019-09965-0
Balaž AM, Stevanović J, Ostafe R, Blažić M, Ilić Đurđić K, Fischer R, Prodanović R. Semi‑rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide. in Molecular Diversity. 2020;24:593-601. doi:10.1007/s11030-019-09965-0 .
Balaž, Ana Marija, Stevanović, Jelena, Ostafe, Raluca, Blažić, Marija, Ilić Đurđić, Karla, Fischer, Rainer, Prodanović, Radivoje, "Semi‑rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide" in Molecular Diversity, 24 (2020):593-601, https://doi.org/10.1007/s11030-019-09965-0 . .