TY  - CONF
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Mutić, Tamara
AU  - Lujić, Tamara
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6524
AB  - Microplastic represents one of the major types of pollutants in modern era. Over several years of research in the field of microplastic, there are still many unknown gaps, including the effects and mechanisms of action of these particles on human health. Studies in this field conducted experiments on cells and human tissues or animals like rats and mice. While these studies suggest the toxic effects of microplastic, it is not clear if concentrations used for exposure are relevant for humans. Also, most of the studies used spherical polystyrene, which does not reflect well the diversity of microplastic particles found in nature. Another gap is lack of studies describing direct interactions of microplastics and proteins. While it is generally known that proteins form corona around microplastic particles, affinity studies and consequences on protein structure are usually missing. The aim of this work was to analyze interaction of a major egg white protein and allergen, ovalbumin to several to microplastic particles, including polystyrene (PS) of 120 and 500 μm in size and polyethylene terephthalate (PET) of 120 μm in size. Binding affinity was determined at both acidic, pH 3 and neutral, pH 7 conditions, at the room temperature, by measuring bulk ovalbumin concentration in supernatants at the equilibrium time. Several binding models, including Langmuir,
Freundlich, Redlich–Peterson and Guggenheim-Anderson-de Boer (GAB), were used to determine binding parameters. The formation of soft and hard corona was analyzed according to the published protocol [1]. Structural analysis was performed using near and far-UV CD spectrometry.
Obtained results showed that ovalbumin binds to both PS and PET. All binding models indicated that ovalbumin binds with higher affinity to tested microplastics on pH 3, compared to pH 7, with the highest affinity being calculated for PS 120 μm. Further analysis showed that ovalbumin forms both soft and hard corona onto the surface of all three microplastics. Structural alterations of ovalbumin as a consequence of its interaction with microplastic was shown to be both pH and
microplastic type dependent. Also, more pronounced effect on its tertiary structure was observed, compared to secondary. At pH3, tertiary structure of bulk ovalbumin becomes destabilized, especially in the presence of PET 120 μm and PS 500 μm, while at pH 7, structural stabilization is observed, especially in the presence of PS 120 μm. Considering that the microplastic was discovered in eggs [2], obtained results suggest that direct interactions of native ovalbumin with microplastic particles could have influence on its structure and thus affect its techno-functional properties.
PB  - Belgrade : Serbian Chemical Society
C3  - Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
T1  - Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions
SP  - 137
EP  - 137
UR  - https://hdl.handle.net/21.15107/rcub_cer_6524
ER  - 

@conference{
author = "Gligorijević, Nikola and Stanić-Vučinić, Dragana and Mutić, Tamara and Lujić, Tamara and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Microplastic represents one of the major types of pollutants in modern era. Over several years of research in the field of microplastic, there are still many unknown gaps, including the effects and mechanisms of action of these particles on human health. Studies in this field conducted experiments on cells and human tissues or animals like rats and mice. While these studies suggest the toxic effects of microplastic, it is not clear if concentrations used for exposure are relevant for humans. Also, most of the studies used spherical polystyrene, which does not reflect well the diversity of microplastic particles found in nature. Another gap is lack of studies describing direct interactions of microplastics and proteins. While it is generally known that proteins form corona around microplastic particles, affinity studies and consequences on protein structure are usually missing. The aim of this work was to analyze interaction of a major egg white protein and allergen, ovalbumin to several to microplastic particles, including polystyrene (PS) of 120 and 500 μm in size and polyethylene terephthalate (PET) of 120 μm in size. Binding affinity was determined at both acidic, pH 3 and neutral, pH 7 conditions, at the room temperature, by measuring bulk ovalbumin concentration in supernatants at the equilibrium time. Several binding models, including Langmuir,
Freundlich, Redlich–Peterson and Guggenheim-Anderson-de Boer (GAB), were used to determine binding parameters. The formation of soft and hard corona was analyzed according to the published protocol [1]. Structural analysis was performed using near and far-UV CD spectrometry.
Obtained results showed that ovalbumin binds to both PS and PET. All binding models indicated that ovalbumin binds with higher affinity to tested microplastics on pH 3, compared to pH 7, with the highest affinity being calculated for PS 120 μm. Further analysis showed that ovalbumin forms both soft and hard corona onto the surface of all three microplastics. Structural alterations of ovalbumin as a consequence of its interaction with microplastic was shown to be both pH and
microplastic type dependent. Also, more pronounced effect on its tertiary structure was observed, compared to secondary. At pH3, tertiary structure of bulk ovalbumin becomes destabilized, especially in the presence of PET 120 μm and PS 500 μm, while at pH 7, structural stabilization is observed, especially in the presence of PS 120 μm. Considering that the microplastic was discovered in eggs [2], obtained results suggest that direct interactions of native ovalbumin with microplastic particles could have influence on its structure and thus affect its techno-functional properties.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023",
title = "Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions",
pages = "137-137",
url = "https://hdl.handle.net/21.15107/rcub_cer_6524"
}

Gligorijević, N., Stanić-Vučinić, D., Mutić, T., Lujić, T.,& Ćirković Veličković, T.. (2023). Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
Belgrade : Serbian Chemical Society., 137-137.
https://hdl.handle.net/21.15107/rcub_cer_6524

Gligorijević N, Stanić-Vučinić D, Mutić T, Lujić T, Ćirković Veličković T. Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023. 2023;:137-137.
https://hdl.handle.net/21.15107/rcub_cer_6524 .

Gligorijević, Nikola, Stanić-Vučinić, Dragana, Mutić, Tamara, Lujić, Tamara, Ćirković Veličković, Tanja, "Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions" in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023 (2023):137-137,
https://hdl.handle.net/21.15107/rcub_cer_6524 .

TY  - CONF
AU  - Lujić, Tamara
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6525
AB  - Ovalbumin (OVA) is the most abundant protein in chicken egg white. It is one of the major allergens in eggs. Micro- and nanoplatic particles (MNPs) are a widespread contaminant and have been found in food and water. It is still unclear how MNPs might affect human health. However, due to their large surface area they have been found to bind various biopolymers,
including proteins. These biopolymers can be bound more strongly or loosely, and are referred to as hard and soft corona, respectfully [1]. MPs have been found in eggs, in the size range of 50-100 μm [2]. It is shown that these particles can interact with proteins and induce structural changes, but there is still not enough information on this topic [3]. These structural changes could lead to a decreased digestibility in the gastrointestinal tract, which could increase the immune
response to known allergens. The aim of this study was to determine whether there are structural changes present in the OVA after incubation with two types of MPs – 120 μm polyethylene terephthalate (PET) and 120 μm polystyrene (PS) and whether they could influence
digestion of OVA with gastrointestinal enzymes. 20 mg of MPs were incubated with 1.3 mg/mL ovalbumin for 4 h at room temperature in a 20 mM phosphate buffer at pH 7. Bulk ovalbumin was separated from the MPs by centrifugation and by filtration through a 0.22 μm PVDF filter. Soft corona was obtained by washing the MPs with water, and the MPs were later removed as described with bulk ovalbumin. Formation of amyloids was monitored with a Thioflavin T (ThT) assay at room temperature and after thermal treatment, and additional structural analysis was performed by circular dichroism (CD) spectrometry in the far-UV region. Thermal stability was also determined by spectrofluorimetry. Digestion with two proteases (pepsin and trypsin) was performed to determine whether there is a change in the gastrointestinal digestibility of OVA.
Results from the ThT assay show that at room temperature there is no significant difference between the fluorescence emission obtained for all samples, with bulk OVA from both MPs showing a slight decrease. However, there is an increase of fluorescence after thermal treatment in all OVA samples, where OVA from the soft corona emits significantly less fluorescence
than control and bulk samples for both types of MPs. Additionally, soft coronas have been shown to have more β-sheet content than other samples, which is more pronounced for OVA incubated with PET. For the heated samples there is a sharp change from α-helix to β-sheets in all the samples, but it is the most dramatic in the soft coronas. This could impose rigidity to the tertiary structure, which would explain why the ThT molecule does not bind as strongly. Despite differences in both the secondary and tertiary structure, the thermal stability is almost the same in all samples. Digestion of the samples shows that the soft corona incubated with PS tends to be more resistant to trypsin than other samples after 2 min, but it is not significant. For digestion with pepsin there is no difference between the samples. In conjunction with the previous results, which indicates a structural stabilisation of the soft corona at pH 7, it is not surprising that there is an increased resistance to trypsin, compared to pepsin which is a gastric enzyme and for which digestion is performed at an acidic pH. In conclusion, there is a structural change present in samples upon contact with MPs, particularly in the soft corona, of which the most pronounced is a decrease of α-helix content and increase in β-sheet content as determined by far-UV CD.
This leads to a structural stabilization which could further impact the digestibility of the OVA protein and impact its allergenicity. However, this must be confirmed with further experiments.
PB  - Belgrade : Serbian Chemical Society
C3  - Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
T1  - Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility
SP  - 153
EP  - 153
UR  - https://hdl.handle.net/21.15107/rcub_cer_6525
ER  - 

@conference{
author = "Lujić, Tamara and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Ovalbumin (OVA) is the most abundant protein in chicken egg white. It is one of the major allergens in eggs. Micro- and nanoplatic particles (MNPs) are a widespread contaminant and have been found in food and water. It is still unclear how MNPs might affect human health. However, due to their large surface area they have been found to bind various biopolymers,
including proteins. These biopolymers can be bound more strongly or loosely, and are referred to as hard and soft corona, respectfully [1]. MPs have been found in eggs, in the size range of 50-100 μm [2]. It is shown that these particles can interact with proteins and induce structural changes, but there is still not enough information on this topic [3]. These structural changes could lead to a decreased digestibility in the gastrointestinal tract, which could increase the immune
response to known allergens. The aim of this study was to determine whether there are structural changes present in the OVA after incubation with two types of MPs – 120 μm polyethylene terephthalate (PET) and 120 μm polystyrene (PS) and whether they could influence
digestion of OVA with gastrointestinal enzymes. 20 mg of MPs were incubated with 1.3 mg/mL ovalbumin for 4 h at room temperature in a 20 mM phosphate buffer at pH 7. Bulk ovalbumin was separated from the MPs by centrifugation and by filtration through a 0.22 μm PVDF filter. Soft corona was obtained by washing the MPs with water, and the MPs were later removed as described with bulk ovalbumin. Formation of amyloids was monitored with a Thioflavin T (ThT) assay at room temperature and after thermal treatment, and additional structural analysis was performed by circular dichroism (CD) spectrometry in the far-UV region. Thermal stability was also determined by spectrofluorimetry. Digestion with two proteases (pepsin and trypsin) was performed to determine whether there is a change in the gastrointestinal digestibility of OVA.
Results from the ThT assay show that at room temperature there is no significant difference between the fluorescence emission obtained for all samples, with bulk OVA from both MPs showing a slight decrease. However, there is an increase of fluorescence after thermal treatment in all OVA samples, where OVA from the soft corona emits significantly less fluorescence
than control and bulk samples for both types of MPs. Additionally, soft coronas have been shown to have more β-sheet content than other samples, which is more pronounced for OVA incubated with PET. For the heated samples there is a sharp change from α-helix to β-sheets in all the samples, but it is the most dramatic in the soft coronas. This could impose rigidity to the tertiary structure, which would explain why the ThT molecule does not bind as strongly. Despite differences in both the secondary and tertiary structure, the thermal stability is almost the same in all samples. Digestion of the samples shows that the soft corona incubated with PS tends to be more resistant to trypsin than other samples after 2 min, but it is not significant. For digestion with pepsin there is no difference between the samples. In conjunction with the previous results, which indicates a structural stabilisation of the soft corona at pH 7, it is not surprising that there is an increased resistance to trypsin, compared to pepsin which is a gastric enzyme and for which digestion is performed at an acidic pH. In conclusion, there is a structural change present in samples upon contact with MPs, particularly in the soft corona, of which the most pronounced is a decrease of α-helix content and increase in β-sheet content as determined by far-UV CD.
This leads to a structural stabilization which could further impact the digestibility of the OVA protein and impact its allergenicity. However, this must be confirmed with further experiments.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023",
title = "Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility",
pages = "153-153",
url = "https://hdl.handle.net/21.15107/rcub_cer_6525"
}

Lujić, T., Gligorijević, N., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2023). Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
Belgrade : Serbian Chemical Society., 153-153.
https://hdl.handle.net/21.15107/rcub_cer_6525

Lujić T, Gligorijević N, Stanić-Vučinić D, Ćirković Veličković T. Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023. 2023;:153-153.
https://hdl.handle.net/21.15107/rcub_cer_6525 .

Lujić, Tamara, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Investigation of structural changes in ovalbumin induced by two types of MPs and its impact on protein digestibility" in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023 (2023):153-153,
https://hdl.handle.net/21.15107/rcub_cer_6525 .

TY  - CONF
AU  - Lujić, Tamara
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Bićanin, Maša
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6651
AB  - Mircoplastics (MPs) are an abundant contaminant in the environment with ingestion being the most common way of exposure for humans. Binding of protein to MPs is proposed to be multilayered with the formation of a soft and hard corona. It has been proven that MPs interact with enzymes present in the digestive system and impact their activity. The aim of this study is to investigate the impact of MPs on the activity of trypsin in simulated intestinal fluid (SIF). For this purpose, two sizes of polypropylene (large – 180-500 μm, small – 63-180 μm) and one size of polyethylene terephthalate (<80 μm) have been studied. Activity in bulk and soft corona was determined in SIF at 405 nm with N-α- Benzoyl-DL-arginine 4-nitroanilide hydrochloride after different times of incubation. Activity in hard corona was determined after 1 h of incubation with the MPs. Although specific activity in the control decreases through time, there is a tendency for all MPs to preserve activity in bulk and soft corona trypsin after 4 h of incubation. Trypsin remains active in the hard corona, with the activity being an order of magnitude lower than in the control, possibly due to significant changes in structure.
PB  - Serbian Biochemical Society
C3  - Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
T1  - Impact of MPs on trypsin activity in simulated intestinal fluid
SP  - 145
EP  - 145
UR  - https://hdl.handle.net/21.15107/rcub_cer_6651
ER  - 

@conference{
author = "Lujić, Tamara and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Bićanin, Maša and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Mircoplastics (MPs) are an abundant contaminant in the environment with ingestion being the most common way of exposure for humans. Binding of protein to MPs is proposed to be multilayered with the formation of a soft and hard corona. It has been proven that MPs interact with enzymes present in the digestive system and impact their activity. The aim of this study is to investigate the impact of MPs on the activity of trypsin in simulated intestinal fluid (SIF). For this purpose, two sizes of polypropylene (large – 180-500 μm, small – 63-180 μm) and one size of polyethylene terephthalate (<80 μm) have been studied. Activity in bulk and soft corona was determined in SIF at 405 nm with N-α- Benzoyl-DL-arginine 4-nitroanilide hydrochloride after different times of incubation. Activity in hard corona was determined after 1 h of incubation with the MPs. Although specific activity in the control decreases through time, there is a tendency for all MPs to preserve activity in bulk and soft corona trypsin after 4 h of incubation. Trypsin remains active in the hard corona, with the activity being an order of magnitude lower than in the control, possibly due to significant changes in structure.",
publisher = "Serbian Biochemical Society",
journal = "Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia",
title = "Impact of MPs on trypsin activity in simulated intestinal fluid",
pages = "145-145",
url = "https://hdl.handle.net/21.15107/rcub_cer_6651"
}

Lujić, T., Gligorijević, N., Stanić-Vučinić, D., Bićanin, M.,& Ćirković Veličković, T.. (2023). Impact of MPs on trypsin activity in simulated intestinal fluid. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia
Serbian Biochemical Society., 145-145.
https://hdl.handle.net/21.15107/rcub_cer_6651

Lujić T, Gligorijević N, Stanić-Vučinić D, Bićanin M, Ćirković Veličković T. Impact of MPs on trypsin activity in simulated intestinal fluid. in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia. 2023;:145-145.
https://hdl.handle.net/21.15107/rcub_cer_6651 .

Lujić, Tamara, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Bićanin, Maša, Ćirković Veličković, Tanja, "Impact of MPs on trypsin activity in simulated intestinal fluid" in Serbian Biochemical Society Twelfth Conference, International scientific meeting, “Biochemistry in Biotechnology,” September 21-23, 2023, Belgrade, Serbia (2023):145-145,
https://hdl.handle.net/21.15107/rcub_cer_6651 .

TY  - CONF
AU  - Gligorijević, Nikola
AU  - Lujić, Tamara
AU  - Mutić, Tamara
AU  - Vasović, Tamara
AU  - de Guzman, Maria Krishna
AU  - Aćimović, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6898
AB  - Ovalbumin (OVA), a main protein of egg white, has characteristic structural fold of a
serpin-family of proteins, propensity to fibril formation and stability to digestion.
Microplastics (MPs) contaminating our food can interact with food proteins in the food
matrix and during digestion. In this study adsorption of OVA to polystyrene (PS) (110 μm
and 260 μm), polyethylene terephthalate (PET) (140 μm) MPs were investigated in acidic
(pH 3) and neutral (pH 7) conditions. Formations of corona on MPs were investigated
using isolated OVA and egg white protein extract comparatively. OVA adsorption depends
on MPs size, polymer chemistry and pH, being highest in acidic pH and higher for PS.
Adsorption of OVA to PS and PET reaches dynamic equilibrium after 4h resulting in
disruption of tertiary structure and formation of hard and soft corona around MPs. Shorter
fragments of OVA populate hard corona, while soft corona exclusively consist of full
length OVA, albeit in its non-native conformation. The conformational changes resemble
those induced by heat treatment with re-arrangement of α-β secondary structures.
Structural changes are striking for the OVA in corona around MPs. Soft corona OVA
preserves thermal and proteolytic stability, but loses ability to form fibrils upon heating.
OVA is abundantly present in corona around MPs also in the presence of other egg white
proteins. MPs contaminating food may bind and change structure and functional properties
of main egg white protein.
PB  - Kragujevac, Srbija : Prirodno-matematički fakultet, Univerzitet u Kragujevcu /  Kragujevac, Serbia : Faculty of Science, University of Kragujevac
C3  - VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs,
T1  - Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate
SP  - OP10
EP  - OP10
UR  - https://hdl.handle.net/21.15107/rcub_cer_6898
ER  - 

@conference{
author = "Gligorijević, Nikola and Lujić, Tamara and Mutić, Tamara and Vasović, Tamara and de Guzman, Maria Krishna and Aćimović, Jelena and Stanić-Vučinić, Dragana and Ćirković Veličković, Tanja",
year = "2023",
abstract = "Ovalbumin (OVA), a main protein of egg white, has characteristic structural fold of a
serpin-family of proteins, propensity to fibril formation and stability to digestion.
Microplastics (MPs) contaminating our food can interact with food proteins in the food
matrix and during digestion. In this study adsorption of OVA to polystyrene (PS) (110 μm
and 260 μm), polyethylene terephthalate (PET) (140 μm) MPs were investigated in acidic
(pH 3) and neutral (pH 7) conditions. Formations of corona on MPs were investigated
using isolated OVA and egg white protein extract comparatively. OVA adsorption depends
on MPs size, polymer chemistry and pH, being highest in acidic pH and higher for PS.
Adsorption of OVA to PS and PET reaches dynamic equilibrium after 4h resulting in
disruption of tertiary structure and formation of hard and soft corona around MPs. Shorter
fragments of OVA populate hard corona, while soft corona exclusively consist of full
length OVA, albeit in its non-native conformation. The conformational changes resemble
those induced by heat treatment with re-arrangement of α-β secondary structures.
Structural changes are striking for the OVA in corona around MPs. Soft corona OVA
preserves thermal and proteolytic stability, but loses ability to form fibrils upon heating.
OVA is abundantly present in corona around MPs also in the presence of other egg white
proteins. MPs contaminating food may bind and change structure and functional properties
of main egg white protein.",
publisher = "Kragujevac, Srbija : Prirodno-matematički fakultet, Univerzitet u Kragujevcu /  Kragujevac, Serbia : Faculty of Science, University of Kragujevac",
journal = "VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs,",
title = "Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate",
pages = "OP10-OP10",
url = "https://hdl.handle.net/21.15107/rcub_cer_6898"
}

Gligorijević, N., Lujić, T., Mutić, T., Vasović, T., de Guzman, M. K., Aćimović, J., Stanić-Vučinić, D.,& Ćirković Veličković, T.. (2023). Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate. in VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs,
Kragujevac, Srbija : Prirodno-matematički fakultet, Univerzitet u Kragujevcu /  Kragujevac, Serbia : Faculty of Science, University of Kragujevac., OP10-OP10.
https://hdl.handle.net/21.15107/rcub_cer_6898

Gligorijević N, Lujić T, Mutić T, Vasović T, de Guzman MK, Aćimović J, Stanić-Vučinić D, Ćirković Veličković T. Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate. in VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs,. 2023;:OP10-OP10.
https://hdl.handle.net/21.15107/rcub_cer_6898 .

Gligorijević, Nikola, Lujić, Tamara, Mutić, Tamara, Vasović, Tamara, de Guzman, Maria Krishna, Aćimović, Jelena, Stanić-Vučinić, Dragana, Ćirković Veličković, Tanja, "Biocorona formation of hen egg white proteins onto the surface of polystyrene and polyethylene terephthalate" in VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike,” Zbornik apstrakata, 2. jun 2023. godine, Kragujevac / VI Symposium of a Serbian proteomic society: „Discussion and Application of  New Methods of Proteomics,“ Book of abstratcs, (2023):OP10-OP10,
https://hdl.handle.net/21.15107/rcub_cer_6898 .

TY  - CONF
AU  - Lujić, Tamara
AU  - Gligorijević, Nikola
AU  - Jovanović, Vesna B.
AU  - Aćimović, Jelena
AU  - Mitić, Dragana
AU  - Vasović, Tamara
AU  - Stojadinović, Marija
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/7473
AB  - Microplastics is abundant in the environment, food and beverages and get ingested by humans.
Its complex interplay with proteins lead to formation of corona. Tightly bound proteins represent
hard corona, while weaker binding partners are found in soft corona. Separation of hard and soft
corona of allergenic proteins of shrimps, eggs and cow’s milk, tropomyosin (TPM), ovalbumin
(OVA) and beta-lactoglobulin (BLG) and identification of binding partners by proteomics was
aim of our study.
Allergenic proteins were purified from egg white, shrimps and cow’s milk. Binding to
polyethylene terephthalate microplastics (PET) (70-100 m) was probed at pH 7 for purified
allergens and egg white proteins. After establishment of binding equilibrium, soft and hard
corona were separated and analyzed by SDS PAGE, followed by identification of bound
proteins by nanoLC-HRMS. Binding of all allergenic proteins was observed in both soft and
hard corona. Soft corona contains exclusively intact, full length OVA, TPM and BLG. Hard
corona is enriched for truncated OVA and oligomers of TPM. OVA fragments are partially or
fully enfolded and have higher level of exposed hydrophobic patches resulting in higher affinity
for PET microplastics. In comparison to OVA and TPM, hard corona of BLG is less abundant
under similar conditions. BLG is compact globular protein with lower level of exposed
hydrophobic patches in comparison to ovalbumin and tropomyosin. In hard corona, trace
amounts of contaminating alfa-lactalbumin become enriched. In the presence of egg white
protein extract OVA forms both SC and HC on microplastics, being the dominant protein of
hard corona (with ovotransferrin). Lysozyme and ovomucin are present only in hard corona.
Both proteins are known for their strong bioactivity and represent a small fraction of total egg
white proteins.
Our results show that allergenic proteins form hard corona on PET microplastics. Among egg
white proteins, minor proteins such as lysozyme and ovomucin become enriched. Denaturing
effect of strong binding to microplastics may change functional characteristics of allergens and
bioactive proteins of foods and should be further investigated in functional assays.
PB  - Italian Proteomics Association
C3  - XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy
T1  - Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics
SP  - 11
EP  - 11
UR  - https://hdl.handle.net/21.15107/rcub_cer_7473
ER  - 

@conference{
author = "Lujić, Tamara and Gligorijević, Nikola and Jovanović, Vesna B. and Aćimović, Jelena and Mitić, Dragana and Vasović, Tamara and Stojadinović, Marija and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Microplastics is abundant in the environment, food and beverages and get ingested by humans.
Its complex interplay with proteins lead to formation of corona. Tightly bound proteins represent
hard corona, while weaker binding partners are found in soft corona. Separation of hard and soft
corona of allergenic proteins of shrimps, eggs and cow’s milk, tropomyosin (TPM), ovalbumin
(OVA) and beta-lactoglobulin (BLG) and identification of binding partners by proteomics was
aim of our study.
Allergenic proteins were purified from egg white, shrimps and cow’s milk. Binding to
polyethylene terephthalate microplastics (PET) (70-100 m) was probed at pH 7 for purified
allergens and egg white proteins. After establishment of binding equilibrium, soft and hard
corona were separated and analyzed by SDS PAGE, followed by identification of bound
proteins by nanoLC-HRMS. Binding of all allergenic proteins was observed in both soft and
hard corona. Soft corona contains exclusively intact, full length OVA, TPM and BLG. Hard
corona is enriched for truncated OVA and oligomers of TPM. OVA fragments are partially or
fully enfolded and have higher level of exposed hydrophobic patches resulting in higher affinity
for PET microplastics. In comparison to OVA and TPM, hard corona of BLG is less abundant
under similar conditions. BLG is compact globular protein with lower level of exposed
hydrophobic patches in comparison to ovalbumin and tropomyosin. In hard corona, trace
amounts of contaminating alfa-lactalbumin become enriched. In the presence of egg white
protein extract OVA forms both SC and HC on microplastics, being the dominant protein of
hard corona (with ovotransferrin). Lysozyme and ovomucin are present only in hard corona.
Both proteins are known for their strong bioactivity and represent a small fraction of total egg
white proteins.
Our results show that allergenic proteins form hard corona on PET microplastics. Among egg
white proteins, minor proteins such as lysozyme and ovomucin become enriched. Denaturing
effect of strong binding to microplastics may change functional characteristics of allergens and
bioactive proteins of foods and should be further investigated in functional assays.",
publisher = "Italian Proteomics Association",
journal = "XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy",
title = "Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics",
pages = "11-11",
url = "https://hdl.handle.net/21.15107/rcub_cer_7473"
}

Lujić, T., Gligorijević, N., Jovanović, V. B., Aćimović, J., Mitić, D., Vasović, T., Stojadinović, M., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2023). Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics. in XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy
Italian Proteomics Association., 11-11.
https://hdl.handle.net/21.15107/rcub_cer_7473

Lujić T, Gligorijević N, Jovanović VB, Aćimović J, Mitić D, Vasović T, Stojadinović M, Stanić-Vučinić D, Ćirković-Veličković T. Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics. in XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy. 2023;:11-11.
https://hdl.handle.net/21.15107/rcub_cer_7473 .

Lujić, Tamara, Gligorijević, Nikola, Jovanović, Vesna B., Aćimović, Jelena, Mitić, Dragana, Vasović, Tamara, Stojadinović, Marija, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics" in XVII International Italian Proteomics Association Annual Meeting in partnership with the Hellenic Proteomics Society and Serbian Proteomics Association, "Proteomics and Metabolomics towards Global Health", November 29th-December 1st, 2023, Ospedale Isola Tiberina - Gemelli Isola,  Roma, Italy (2023):11-11,
https://hdl.handle.net/21.15107/rcub_cer_7473 .

TY  - JOUR
AU  - de Guzman, Maria Krishna
AU  - Stanić-Vučinić, Dragana
AU  - Gligorijević, Nikola
AU  - Wimmer, Lukas
AU  - Gasparyan, Manvel
AU  - Lujić, Tamara
AU  - Vasović, Tamara
AU  - Dailey, Lea Ann
AU  - Van Haute, Sam
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/6466
AB  - Human ingestion of microplastics (MPs) is common and inevitable due to the widespread contamination of food items, but implications on the gastric digestion of food proteins are still unknown. In this study, the interactions between pepsin and polystyrene (PS) MPs were evaluated by investigating enzyme activity and conformation in a simulated human gastric environment in the presence or absence of PS MPs. The impact on food digestion was also assessed by monitoring the kinetics of protein hydrolysis through static in vitro gastric digestion of cow's milk contaminated with PS. The binding of pepsin to PS showed that the surface chemistry of MPs dictates binding affinity. The key contributor to pepsin adsorption seems to be π−π interactions between the aromatic residues and the PS phenyl rings. During quick exposure (10 min) of pepsin to increasing concentrations (222, 2219, 22188 particles/mL) of 10 μm PS (PS10) and 100 μm PS (PS100), total enzymatic activities were not affected remarkably. However, upon prolonged exposure at 1 and 2 h, preferential binding of pepsin to the small, low zeta-potential PS caused structural changes in the protein which led to a significant reduction of its activity. Digestion of cow's milk mixed with PS10 resulted in transient accumulation of larger peptides (10–35 kDa) and reduced bioavailability of short peptides (2–9 kDa) in the gastric phase. This, however, was only observed at extremely high PS10 concentration (0.3 mg/mL or 5.46E+05 particles/mL). The digestion of milk peptides, bound preferentially over pepsin within the hard corona on the PS10 surface, was delayed up to 15 min in comparison to bulk protein digestion. Intact caseins, otherwise rapidly digested, remained bound to PS10 in the hard corona for up to 15 min. This work presents valuable insights regarding the interaction of MPs, food proteins, and pepsin, and their dynamics during gastric digestion.
PB  - Elsevier Ltd.
T2  - Environmental Pollution
T1  - Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion
VL  - 335
SP  - 122282
DO  - 10.1016/j.envpol.2023.122282
ER  - 

@article{
author = "de Guzman, Maria Krishna and Stanić-Vučinić, Dragana and Gligorijević, Nikola and Wimmer, Lukas and Gasparyan, Manvel and Lujić, Tamara and Vasović, Tamara and Dailey, Lea Ann and Van Haute, Sam and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Human ingestion of microplastics (MPs) is common and inevitable due to the widespread contamination of food items, but implications on the gastric digestion of food proteins are still unknown. In this study, the interactions between pepsin and polystyrene (PS) MPs were evaluated by investigating enzyme activity and conformation in a simulated human gastric environment in the presence or absence of PS MPs. The impact on food digestion was also assessed by monitoring the kinetics of protein hydrolysis through static in vitro gastric digestion of cow's milk contaminated with PS. The binding of pepsin to PS showed that the surface chemistry of MPs dictates binding affinity. The key contributor to pepsin adsorption seems to be π−π interactions between the aromatic residues and the PS phenyl rings. During quick exposure (10 min) of pepsin to increasing concentrations (222, 2219, 22188 particles/mL) of 10 μm PS (PS10) and 100 μm PS (PS100), total enzymatic activities were not affected remarkably. However, upon prolonged exposure at 1 and 2 h, preferential binding of pepsin to the small, low zeta-potential PS caused structural changes in the protein which led to a significant reduction of its activity. Digestion of cow's milk mixed with PS10 resulted in transient accumulation of larger peptides (10–35 kDa) and reduced bioavailability of short peptides (2–9 kDa) in the gastric phase. This, however, was only observed at extremely high PS10 concentration (0.3 mg/mL or 5.46E+05 particles/mL). The digestion of milk peptides, bound preferentially over pepsin within the hard corona on the PS10 surface, was delayed up to 15 min in comparison to bulk protein digestion. Intact caseins, otherwise rapidly digested, remained bound to PS10 in the hard corona for up to 15 min. This work presents valuable insights regarding the interaction of MPs, food proteins, and pepsin, and their dynamics during gastric digestion.",
publisher = "Elsevier Ltd.",
journal = "Environmental Pollution",
title = "Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion",
volume = "335",
pages = "122282",
doi = "10.1016/j.envpol.2023.122282"
}

de Guzman, M. K., Stanić-Vučinić, D., Gligorijević, N., Wimmer, L., Gasparyan, M., Lujić, T., Vasović, T., Dailey, L. A., Van Haute, S.,& Ćirković-Veličković, T.. (2023). Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution
Elsevier Ltd.., 335, 122282.
https://doi.org/10.1016/j.envpol.2023.122282

de Guzman MK, Stanić-Vučinić D, Gligorijević N, Wimmer L, Gasparyan M, Lujić T, Vasović T, Dailey LA, Van Haute S, Ćirković-Veličković T. Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution. 2023;335:122282.
doi:10.1016/j.envpol.2023.122282 .

de Guzman, Maria Krishna, Stanić-Vučinić, Dragana, Gligorijević, Nikola, Wimmer, Lukas, Gasparyan, Manvel, Lujić, Tamara, Vasović, Tamara, Dailey, Lea Ann, Van Haute, Sam, Ćirković-Veličković, Tanja, "Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion" in Environmental Pollution, 335 (2023):122282,
https://doi.org/10.1016/j.envpol.2023.122282 . .