STUDY AND PREPARATION OF ARTIFICIAL MANGANESE METALLOENZYMES WITH LACCASE LIKE ACTIVITY
Samo za registrovane korisnike
2023
Autori
García-Sanz, CarlaAndreu, Alicia
de las Rivas, Blanca
Muñoz, Rosario
Vukoičić, Ana
Milivojević, Ana
Bezbradica, Dejan
Palomo, Jose Miguel
Konferencijski prilog (Objavljena verzija)
Metapodaci
Prikaz svih podataka o dokumentuApstrakt
Laccase (EC 1.10.3.2, p-diphenol: dioxygen oxidoreductase) is a blue copper oxidase
of great industrial interest due to its ability to catalyseoxidation processes of phenols and
persistent organic pollutants. However, it is susceptible to denaturation by high temperatures,
and sensitive to pH and the presence of high concentrations of solvents, which is a problem
for industrial use. To solve this problem, this project develops the synthesis in aqueous
medium of a new Mn metalloenzyme with laccase oxidase mimetic catalytic activity. To do
this, Geobacillus thermocatenulatus lipase (GTL) is used as a "scaffold" enzyme, which is
mixed with a manganese salt at 50 ºC in an aqueous medium. In this way, the in situ
formation of manganese (IV) oxide nanowires is generated, interacting with the enzyme and
obtaining the GTL-Mn bioconjugate. On the other hand, its oxidative activity was evaluated
using the ABTS assay, obtaining a specificity 300 times greater than the laccase from
Tram...etes versicolor and 2 times more than the laccase from Myceliophthora thermophila
expressed in Aspergillus oryzae (Novozym 51003®). In addition, the new metalloenzyme
turned out to be 2 times more stable at 40 ºC, 3 times more stable in thepresence of 10% can,
and 10 times more stable at 20% and 30% AcN than laccase (Novozym 51003®) by
evaluating it at 2 hour incubation. Moreover, it was shown that the use of immobilization
strategies improves the stabilization of this artificial metalloenzyme two times more. Finally,
the novel Mn metalloenzymes were effective in the reaction of phlorizin oligomerization.
Future research will aim to extend this study for the use of flavonoids as prebiotic molecules
in the cosmetics industry.
Ključne reči:
metalloenzyme / Mn / laccase like activity / oxidantIzvor:
Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade, 2023, 35-Izdavač:
- Belgrade : University, Faculty of Technology and Metallurgy
Finansiranje / projekti:
- Spanish National Research Council (CSIC)
- Ministarstvo nauke, tehnološkog razvoja i inovacija Republike Srbije, institucionalno finansiranje - 200135 (Univerzitet u Beogradu, Tehnološko-metalurški fakultet) (RS-MESTD-inst-2020-200135)
- Ministarstvo nauke, tehnološkog razvoja i inovacija Republike Srbije, institucionalno finansiranje - 200287 (Inovacioni centar Tehnološko-metalurškog fakulteta u Beogradu doo) (RS-MESTD-inst-2020-200287)
- European Commission, Horizon Europe 2021-2027 research and innovation programme under grant agreement ID 101060130 (TwinPrebioEnz)
- COST Action CA18132 (GlycoNanoProbes)
Kolekcije
Institucija/grupa
Inovacioni centarTY - CONF AU - García-Sanz, Carla AU - Andreu, Alicia AU - de las Rivas, Blanca AU - Muñoz, Rosario AU - Vukoičić, Ana AU - Milivojević, Ana AU - Bezbradica, Dejan AU - Palomo, Jose Miguel PY - 2023 UR - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/6949 AB - Laccase (EC 1.10.3.2, p-diphenol: dioxygen oxidoreductase) is a blue copper oxidase of great industrial interest due to its ability to catalyseoxidation processes of phenols and persistent organic pollutants. However, it is susceptible to denaturation by high temperatures, and sensitive to pH and the presence of high concentrations of solvents, which is a problem for industrial use. To solve this problem, this project develops the synthesis in aqueous medium of a new Mn metalloenzyme with laccase oxidase mimetic catalytic activity. To do this, Geobacillus thermocatenulatus lipase (GTL) is used as a "scaffold" enzyme, which is mixed with a manganese salt at 50 ºC in an aqueous medium. In this way, the in situ formation of manganese (IV) oxide nanowires is generated, interacting with the enzyme and obtaining the GTL-Mn bioconjugate. On the other hand, its oxidative activity was evaluated using the ABTS assay, obtaining a specificity 300 times greater than the laccase from Trametes versicolor and 2 times more than the laccase from Myceliophthora thermophila expressed in Aspergillus oryzae (Novozym 51003®). In addition, the new metalloenzyme turned out to be 2 times more stable at 40 ºC, 3 times more stable in thepresence of 10% can, and 10 times more stable at 20% and 30% AcN than laccase (Novozym 51003®) by evaluating it at 2 hour incubation. Moreover, it was shown that the use of immobilization strategies improves the stabilization of this artificial metalloenzyme two times more. Finally, the novel Mn metalloenzymes were effective in the reaction of phlorizin oligomerization. Future research will aim to extend this study for the use of flavonoids as prebiotic molecules in the cosmetics industry. PB - Belgrade : University, Faculty of Technology and Metallurgy C3 - Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade T1 - STUDY AND PREPARATION OF ARTIFICIAL MANGANESE METALLOENZYMES WITH LACCASE LIKE ACTIVITY SP - 35 UR - https://hdl.handle.net/21.15107/rcub_technorep_6949 ER -
@conference{ author = "García-Sanz, Carla and Andreu, Alicia and de las Rivas, Blanca and Muñoz, Rosario and Vukoičić, Ana and Milivojević, Ana and Bezbradica, Dejan and Palomo, Jose Miguel", year = "2023", abstract = "Laccase (EC 1.10.3.2, p-diphenol: dioxygen oxidoreductase) is a blue copper oxidase of great industrial interest due to its ability to catalyseoxidation processes of phenols and persistent organic pollutants. However, it is susceptible to denaturation by high temperatures, and sensitive to pH and the presence of high concentrations of solvents, which is a problem for industrial use. To solve this problem, this project develops the synthesis in aqueous medium of a new Mn metalloenzyme with laccase oxidase mimetic catalytic activity. To do this, Geobacillus thermocatenulatus lipase (GTL) is used as a "scaffold" enzyme, which is mixed with a manganese salt at 50 ºC in an aqueous medium. In this way, the in situ formation of manganese (IV) oxide nanowires is generated, interacting with the enzyme and obtaining the GTL-Mn bioconjugate. On the other hand, its oxidative activity was evaluated using the ABTS assay, obtaining a specificity 300 times greater than the laccase from Trametes versicolor and 2 times more than the laccase from Myceliophthora thermophila expressed in Aspergillus oryzae (Novozym 51003®). In addition, the new metalloenzyme turned out to be 2 times more stable at 40 ºC, 3 times more stable in thepresence of 10% can, and 10 times more stable at 20% and 30% AcN than laccase (Novozym 51003®) by evaluating it at 2 hour incubation. Moreover, it was shown that the use of immobilization strategies improves the stabilization of this artificial metalloenzyme two times more. Finally, the novel Mn metalloenzymes were effective in the reaction of phlorizin oligomerization. Future research will aim to extend this study for the use of flavonoids as prebiotic molecules in the cosmetics industry.", publisher = "Belgrade : University, Faculty of Technology and Metallurgy", journal = "Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade", title = "STUDY AND PREPARATION OF ARTIFICIAL MANGANESE METALLOENZYMES WITH LACCASE LIKE ACTIVITY", pages = "35", url = "https://hdl.handle.net/21.15107/rcub_technorep_6949" }
García-Sanz, C., Andreu, A., de las Rivas, B., Muñoz, R., Vukoičić, A., Milivojević, A., Bezbradica, D.,& Palomo, J. M.. (2023). STUDY AND PREPARATION OF ARTIFICIAL MANGANESE METALLOENZYMES WITH LACCASE LIKE ACTIVITY. in Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade Belgrade : University, Faculty of Technology and Metallurgy., 35. https://hdl.handle.net/21.15107/rcub_technorep_6949
García-Sanz C, Andreu A, de las Rivas B, Muñoz R, Vukoičić A, Milivojević A, Bezbradica D, Palomo JM. STUDY AND PREPARATION OF ARTIFICIAL MANGANESE METALLOENZYMES WITH LACCASE LIKE ACTIVITY. in Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade. 2023;:35. https://hdl.handle.net/21.15107/rcub_technorep_6949 .
García-Sanz, Carla, Andreu, Alicia, de las Rivas, Blanca, Muñoz, Rosario, Vukoičić, Ana, Milivojević, Ana, Bezbradica, Dejan, Palomo, Jose Miguel, "STUDY AND PREPARATION OF ARTIFICIAL MANGANESE METALLOENZYMES WITH LACCASE LIKE ACTIVITY" in Book of Abstracts / International Conference Biochemical Engineering and Biotechnology for Young Scientists, 7-8 December, 2023, Belgrade (2023):35, https://hdl.handle.net/21.15107/rcub_technorep_6949 .