Semi-rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide
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2020
Authors
Balaž, Ana Marija
Stevanović, Jelena
Ostafe, Raluca

Blazić, Marija
Ilić Đurđić, Karla

Fischer, Rainer

Prodanović, Radivoje

Article (Published version)

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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.
Keywords:
Protein engineering / Hydrogen peroxide / Library / Methionine / YeastSource:
Molecular Diversity, 2020, 24, 3, 593-601Publisher:
- Springer
Funding / projects:
- Novel encapsulation and enzyme technologies for designing of new biocatalysts and biologically active compounds targeting enhancement of food quality, safety and competitiveness (RS-46010)
- Allergens, antibodies, enzymes and small physiologically important molecules: design, structure, function and relevance (RS-172049)
- Study of structure-function relationships in the plant cell wall and modifications of the wall structure by enzyme engineering (RS-173017)
DOI: 10.1007/s11030-019-09965-0
ISSN: 1381-1991
WoS: 000547921900001
Scopus: 2-s2.0-85066612562
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Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Balaž, Ana Marija AU - Stevanović, Jelena AU - Ostafe, Raluca AU - Blazić, Marija AU - Ilić Đurđić, Karla AU - Fischer, Rainer AU - Prodanović, Radivoje PY - 2020 UR - https://cherry.chem.bg.ac.rs/handle/123456789/4011 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. T2 - Molecular Diversity T1 - Semi-rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide VL - 24 IS - 3 SP - 593 EP - 601 DO - 10.1007/s11030-019-09965-0 ER -
@article{ author = "Balaž, Ana Marija and Stevanović, Jelena and Ostafe, Raluca and Blazić, 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.", journal = "Molecular Diversity", title = "Semi-rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide", volume = "24", number = "3", pages = "593-601", doi = "10.1007/s11030-019-09965-0" }
Balaž, A. M., Stevanović, J., Ostafe, R., Blazić, 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, 24(3), 593-601. https://doi.org/10.1007/s11030-019-09965-0
Balaž AM, Stevanović J, Ostafe R, Blazić 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(3):593-601. doi:10.1007/s11030-019-09965-0 .
Balaž, Ana Marija, Stevanović, Jelena, Ostafe, Raluca, Blazić, 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, no. 3 (2020):593-601, https://doi.org/10.1007/s11030-019-09965-0 . .