Selective Immobilization of Fructosyltransferase onto Glutaraldehyde Modified Support and Its Application in the Production of Fructo-oligosaccharides
2022
Преузимање 🢃
Аутори
Veljković, MilicaSimović, Milica
Ćorović, Marija
Milivojević, Ana
Petrov Ivanković, Anja
Banjanac, Katarina
Bezbradica, Dejan
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
In recent decades, the scientific community has recognized the growing importance of prebiotics, and therefore, numerous studies are focused on their economical production due to their low presence in natural resources. It has been confirmed that prebiotics are a source of energy for probiotics in the gastrointestinal tract (GIT) and enable their proliferation, consequently leading to the normal functioning of the intestinal microbiota. Also, products of their fermentation are short-chain fatty acids (SCFA), which play a key role in maintaining and improving the health not only of the GIT but also of the whole organism. Among several confirmed prebiotics, fructo-oligosaccharides (FOS) are considered interesting candidates for use in a wide range of products in the food industry. They are characterized as low-calorie and non-cariogenic substances that represent an adequate sugar substitute and can be considered suitable for use in products intended for diabetics. The subject of this res...earch will be the production of FOS by transforming sucrose using a fructosyltransferase (FTase) present in commercial preparation Pectinex® Ultra SP-L, with special emphasis on the development of adequate FTase immobilization method that would enable selective isolation of the enzyme responsible for the synthesis of FOS from the complex enzymatic mixture. This would lead to considerable enzyme purification and allow its direct incorporation into different sucrosebased products, without the fear that action of the other hydrolytic enzymes may adversely affect the products functional characteristics. Accordingly, the possibility of selective immobilization of the enzyme using support with primary amino groups, Purolite® A109, which was previously activated and modified using glutaraldehyde (GA), was investigated. In the initial phase of the research, the effects of individual immobilization parameters such as pH, enzyme concentration and immobilization time were investigated to optimize the process using support chemically activated with 15% and 0.5% GA to form dimers and monomers, respectively. It was determined that highly active immobilized preparations (371.8 IU/g of support - dimer and 213.8 IU/g of support – monomer) were achieved under acidic conditions (pH 4) provided that an enzyme concentration was 50 mg/g of support after 7 h and 3 h, respectively. Bearing in mind the obtained results of the expressed activity, it is noticeable that the formation of dimers showed higher reactivity compared to the form of monomers. Also, in the case of support modification using 15% GA, the value of the ratio of FTase and pectinase (as dominant enzyme mixture component) activity immobilization yields was 16.45, indicating the high feasibility of selective immobilization of FTase on modified polystyrene resin. After obtaining immobilized preparations of satisfactory features, they were tested in a reaction of FOS synthesis under determined optimal conditions. The maximum FOS yields of approximately 50% of total carbohydrates in the reaction mixture were recorded after 21 h. Finally, it can be concluded that the examined immobilization method yielded highly active, stable and more importantly refined enzyme preparation that can be further utilized on a larger scale for development of continual processes for FOS synthesis, as well as for modification of different sucrose-based mediums.
Кључне речи:
Chemical modification / fructo-oligosaccharides / glutaraldehyde / immobilization of fructosyltransferaseИзвор:
XVI. international conference on advances in probiotics and prebiotics, Copenhagen Denmark, Proceedings, 2022, 83-Издавач:
- The world academy of science, engineering and technology
Финансирање / пројекти:
- PrIntPrEnzy - Prebiotics for functional food and bioactive cosmetics produced in intensified enzymatic processes (RS-ScienceFundRS-Ideje-7750109)
Колекције
Институција/група
Inovacioni centarTY - CONF AU - Veljković, Milica AU - Simović, Milica AU - Ćorović, Marija AU - Milivojević, Ana AU - Petrov Ivanković, Anja AU - Banjanac, Katarina AU - Bezbradica, Dejan PY - 2022 UR - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/5995 AB - In recent decades, the scientific community has recognized the growing importance of prebiotics, and therefore, numerous studies are focused on their economical production due to their low presence in natural resources. It has been confirmed that prebiotics are a source of energy for probiotics in the gastrointestinal tract (GIT) and enable their proliferation, consequently leading to the normal functioning of the intestinal microbiota. Also, products of their fermentation are short-chain fatty acids (SCFA), which play a key role in maintaining and improving the health not only of the GIT but also of the whole organism. Among several confirmed prebiotics, fructo-oligosaccharides (FOS) are considered interesting candidates for use in a wide range of products in the food industry. They are characterized as low-calorie and non-cariogenic substances that represent an adequate sugar substitute and can be considered suitable for use in products intended for diabetics. The subject of this research will be the production of FOS by transforming sucrose using a fructosyltransferase (FTase) present in commercial preparation Pectinex® Ultra SP-L, with special emphasis on the development of adequate FTase immobilization method that would enable selective isolation of the enzyme responsible for the synthesis of FOS from the complex enzymatic mixture. This would lead to considerable enzyme purification and allow its direct incorporation into different sucrosebased products, without the fear that action of the other hydrolytic enzymes may adversely affect the products functional characteristics. Accordingly, the possibility of selective immobilization of the enzyme using support with primary amino groups, Purolite® A109, which was previously activated and modified using glutaraldehyde (GA), was investigated. In the initial phase of the research, the effects of individual immobilization parameters such as pH, enzyme concentration and immobilization time were investigated to optimize the process using support chemically activated with 15% and 0.5% GA to form dimers and monomers, respectively. It was determined that highly active immobilized preparations (371.8 IU/g of support - dimer and 213.8 IU/g of support – monomer) were achieved under acidic conditions (pH 4) provided that an enzyme concentration was 50 mg/g of support after 7 h and 3 h, respectively. Bearing in mind the obtained results of the expressed activity, it is noticeable that the formation of dimers showed higher reactivity compared to the form of monomers. Also, in the case of support modification using 15% GA, the value of the ratio of FTase and pectinase (as dominant enzyme mixture component) activity immobilization yields was 16.45, indicating the high feasibility of selective immobilization of FTase on modified polystyrene resin. After obtaining immobilized preparations of satisfactory features, they were tested in a reaction of FOS synthesis under determined optimal conditions. The maximum FOS yields of approximately 50% of total carbohydrates in the reaction mixture were recorded after 21 h. Finally, it can be concluded that the examined immobilization method yielded highly active, stable and more importantly refined enzyme preparation that can be further utilized on a larger scale for development of continual processes for FOS synthesis, as well as for modification of different sucrose-based mediums. PB - The world academy of science, engineering and technology C3 - XVI. international conference on advances in probiotics and prebiotics, Copenhagen Denmark, Proceedings T1 - Selective Immobilization of Fructosyltransferase onto Glutaraldehyde Modified Support and Its Application in the Production of Fructo-oligosaccharides SP - 83 UR - https://hdl.handle.net/21.15107/rcub_technorep_5995 ER -
@conference{ author = "Veljković, Milica and Simović, Milica and Ćorović, Marija and Milivojević, Ana and Petrov Ivanković, Anja and Banjanac, Katarina and Bezbradica, Dejan", year = "2022", abstract = "In recent decades, the scientific community has recognized the growing importance of prebiotics, and therefore, numerous studies are focused on their economical production due to their low presence in natural resources. It has been confirmed that prebiotics are a source of energy for probiotics in the gastrointestinal tract (GIT) and enable their proliferation, consequently leading to the normal functioning of the intestinal microbiota. Also, products of their fermentation are short-chain fatty acids (SCFA), which play a key role in maintaining and improving the health not only of the GIT but also of the whole organism. Among several confirmed prebiotics, fructo-oligosaccharides (FOS) are considered interesting candidates for use in a wide range of products in the food industry. They are characterized as low-calorie and non-cariogenic substances that represent an adequate sugar substitute and can be considered suitable for use in products intended for diabetics. The subject of this research will be the production of FOS by transforming sucrose using a fructosyltransferase (FTase) present in commercial preparation Pectinex® Ultra SP-L, with special emphasis on the development of adequate FTase immobilization method that would enable selective isolation of the enzyme responsible for the synthesis of FOS from the complex enzymatic mixture. This would lead to considerable enzyme purification and allow its direct incorporation into different sucrosebased products, without the fear that action of the other hydrolytic enzymes may adversely affect the products functional characteristics. Accordingly, the possibility of selective immobilization of the enzyme using support with primary amino groups, Purolite® A109, which was previously activated and modified using glutaraldehyde (GA), was investigated. In the initial phase of the research, the effects of individual immobilization parameters such as pH, enzyme concentration and immobilization time were investigated to optimize the process using support chemically activated with 15% and 0.5% GA to form dimers and monomers, respectively. It was determined that highly active immobilized preparations (371.8 IU/g of support - dimer and 213.8 IU/g of support – monomer) were achieved under acidic conditions (pH 4) provided that an enzyme concentration was 50 mg/g of support after 7 h and 3 h, respectively. Bearing in mind the obtained results of the expressed activity, it is noticeable that the formation of dimers showed higher reactivity compared to the form of monomers. Also, in the case of support modification using 15% GA, the value of the ratio of FTase and pectinase (as dominant enzyme mixture component) activity immobilization yields was 16.45, indicating the high feasibility of selective immobilization of FTase on modified polystyrene resin. After obtaining immobilized preparations of satisfactory features, they were tested in a reaction of FOS synthesis under determined optimal conditions. The maximum FOS yields of approximately 50% of total carbohydrates in the reaction mixture were recorded after 21 h. Finally, it can be concluded that the examined immobilization method yielded highly active, stable and more importantly refined enzyme preparation that can be further utilized on a larger scale for development of continual processes for FOS synthesis, as well as for modification of different sucrose-based mediums.", publisher = "The world academy of science, engineering and technology", journal = "XVI. international conference on advances in probiotics and prebiotics, Copenhagen Denmark, Proceedings", title = "Selective Immobilization of Fructosyltransferase onto Glutaraldehyde Modified Support and Its Application in the Production of Fructo-oligosaccharides", pages = "83", url = "https://hdl.handle.net/21.15107/rcub_technorep_5995" }
Veljković, M., Simović, M., Ćorović, M., Milivojević, A., Petrov Ivanković, A., Banjanac, K.,& Bezbradica, D.. (2022). Selective Immobilization of Fructosyltransferase onto Glutaraldehyde Modified Support and Its Application in the Production of Fructo-oligosaccharides. in XVI. international conference on advances in probiotics and prebiotics, Copenhagen Denmark, Proceedings The world academy of science, engineering and technology., 83. https://hdl.handle.net/21.15107/rcub_technorep_5995
Veljković M, Simović M, Ćorović M, Milivojević A, Petrov Ivanković A, Banjanac K, Bezbradica D. Selective Immobilization of Fructosyltransferase onto Glutaraldehyde Modified Support and Its Application in the Production of Fructo-oligosaccharides. in XVI. international conference on advances in probiotics and prebiotics, Copenhagen Denmark, Proceedings. 2022;:83. https://hdl.handle.net/21.15107/rcub_technorep_5995 .
Veljković, Milica, Simović, Milica, Ćorović, Marija, Milivojević, Ana, Petrov Ivanković, Anja, Banjanac, Katarina, Bezbradica, Dejan, "Selective Immobilization of Fructosyltransferase onto Glutaraldehyde Modified Support and Its Application in the Production of Fructo-oligosaccharides" in XVI. international conference on advances in probiotics and prebiotics, Copenhagen Denmark, Proceedings (2022):83, https://hdl.handle.net/21.15107/rcub_technorep_5995 .