TY  - CONF
AU  - Vukoičić, Ana
AU  - Milivojević, Ana
AU  - Banjanac, Katarina
AU  - Petrov Ivanković, Anja
AU  - Ćorović, Marija
AU  - Marinković, Aleksandar
AU  - Palomo, Jose Miguel
AU  - Bezbradica, Dejan
PY  - 2023
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/6844
AB  - Flavonoids are a big class of plants’ secondary metabolites which possess antimicrobial, antioxidative, anti-inflammatory, etc. activities. However, their wider application is limited due to their poor solubility and stability in different environments. In order to overcome this problem and improve their biological and physicochemical characteristics, enzymatic oligomerization of flavonoids can be performed by using laccases as biocatalyst. In this study, laccase from Trametes versicolor was successfully immobilized onto two nanocarriers and used as a catalyst for the oligomerization of two flavonoids, esculin and phloridzin. For immobilization, nanocelulose prepared by ultrasonic treatment of commercial microcellulose and nanosilica from rice were used. Both carriers were modified in order to introduce amino groups on their surface: nanocelulose with poly(ethyleneimine) (NC-PEI) and nanosilica with organosylane (NS-OS). Most active preparation for NC-PEIlaccase and NS-OS-laccase showed activities of 10.05 IU/g of support and 9.08 IU/g of support, respectively. The oligomerization of flavonoids was performed using immobilized laccase and obtained results were compared with those obtained with free laccase (same amount of activity units) as biocatalyst. Immobilized preparations showed promising activities in reaction of esculin oligomerization, since after 24h of reaction the conversions of 73.3% and 61.6% with NC-PEI-laccase and NS-OS-laccase, respectively was achieved, while the soluble enzyme gave the conversion of 92.4%. On the other hand, some lower activity of NC-PEI-laccase was observed in reaction of phloridzin oligomerization, since the achieved conversion was 43.3% compared to 80.7% of phloridzin conversion obtained with soluble laccase. The lower activity could be explained by different reaction medium, since, compared to the esculin oligomerization, the phloridzin oligomerization was performed in 30% methanol, due to its low water solubility. However, obtained results indicate that novel immobilized laccase preparations can be successfully applied in the production of various flavonoid oligomers.
PB  - Belgrade : University, Faculty of Geography
C3  - Book of Abstracts / International Scientific Conference Green Agenda for Western Balkans, Belgrade, June 20-22, 2023
T1  - Immobilization of laccase from Trametes versicolor on different nanocarriers and its application in the synthesis of flavonoid oligomers
SP  - 27
UR  - https://hdl.handle.net/21.15107/rcub_technorep_6844
ER  - 

@conference{
author = "Vukoičić, Ana and Milivojević, Ana and Banjanac, Katarina and Petrov Ivanković, Anja and Ćorović, Marija and Marinković, Aleksandar and Palomo, Jose Miguel and Bezbradica, Dejan",
year = "2023",
abstract = "Flavonoids are a big class of plants’ secondary metabolites which possess antimicrobial, antioxidative, anti-inflammatory, etc. activities. However, their wider application is limited due to their poor solubility and stability in different environments. In order to overcome this problem and improve their biological and physicochemical characteristics, enzymatic oligomerization of flavonoids can be performed by using laccases as biocatalyst. In this study, laccase from Trametes versicolor was successfully immobilized onto two nanocarriers and used as a catalyst for the oligomerization of two flavonoids, esculin and phloridzin. For immobilization, nanocelulose prepared by ultrasonic treatment of commercial microcellulose and nanosilica from rice were used. Both carriers were modified in order to introduce amino groups on their surface: nanocelulose with poly(ethyleneimine) (NC-PEI) and nanosilica with organosylane (NS-OS). Most active preparation for NC-PEIlaccase and NS-OS-laccase showed activities of 10.05 IU/g of support and 9.08 IU/g of support, respectively. The oligomerization of flavonoids was performed using immobilized laccase and obtained results were compared with those obtained with free laccase (same amount of activity units) as biocatalyst. Immobilized preparations showed promising activities in reaction of esculin oligomerization, since after 24h of reaction the conversions of 73.3% and 61.6% with NC-PEI-laccase and NS-OS-laccase, respectively was achieved, while the soluble enzyme gave the conversion of 92.4%. On the other hand, some lower activity of NC-PEI-laccase was observed in reaction of phloridzin oligomerization, since the achieved conversion was 43.3% compared to 80.7% of phloridzin conversion obtained with soluble laccase. The lower activity could be explained by different reaction medium, since, compared to the esculin oligomerization, the phloridzin oligomerization was performed in 30% methanol, due to its low water solubility. However, obtained results indicate that novel immobilized laccase preparations can be successfully applied in the production of various flavonoid oligomers.",
publisher = "Belgrade : University, Faculty of Geography",
journal = "Book of Abstracts / International Scientific Conference Green Agenda for Western Balkans, Belgrade, June 20-22, 2023",
title = "Immobilization of laccase from Trametes versicolor on different nanocarriers and its application in the synthesis of flavonoid oligomers",
pages = "27",
url = "https://hdl.handle.net/21.15107/rcub_technorep_6844"
}

Vukoičić, A., Milivojević, A., Banjanac, K., Petrov Ivanković, A., Ćorović, M., Marinković, A., Palomo, J. M.,& Bezbradica, D.. (2023). Immobilization of laccase from Trametes versicolor on different nanocarriers and its application in the synthesis of flavonoid oligomers. in Book of Abstracts / International Scientific Conference Green Agenda for Western Balkans, Belgrade, June 20-22, 2023
Belgrade : University, Faculty of Geography., 27.
https://hdl.handle.net/21.15107/rcub_technorep_6844

Vukoičić A, Milivojević A, Banjanac K, Petrov Ivanković A, Ćorović M, Marinković A, Palomo JM, Bezbradica D. Immobilization of laccase from Trametes versicolor on different nanocarriers and its application in the synthesis of flavonoid oligomers. in Book of Abstracts / International Scientific Conference Green Agenda for Western Balkans, Belgrade, June 20-22, 2023. 2023;:27.
https://hdl.handle.net/21.15107/rcub_technorep_6844 .

Vukoičić, Ana, Milivojević, Ana, Banjanac, Katarina, Petrov Ivanković, Anja, Ćorović, Marija, Marinković, Aleksandar, Palomo, Jose Miguel, Bezbradica, Dejan, "Immobilization of laccase from Trametes versicolor on different nanocarriers and its application in the synthesis of flavonoid oligomers" in Book of Abstracts / International Scientific Conference Green Agenda for Western Balkans, Belgrade, June 20-22, 2023 (2023):27,
https://hdl.handle.net/21.15107/rcub_technorep_6844 .

TY  - 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 .