Innovation Project / LAVGLU - Innovative processes of production cereals-based functional products enriched with non-allergenic proteins and bioactive peptides

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Innovation Project / LAVGLU - Innovative processes of production cereals-based functional products enriched with non-allergenic proteins and bioactive peptides

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Publications

Optimization of submerged fermentation conditions for gluten-degrading enzyme production using B. subtilis isolate

Gazikalović, Ivana; Jovanović, Jelena; Šekuljica, Nataša; Luković, Nevena; Jakovetić Tanasković, Sonja; Knežević-Jugović, Zorica

(Skopje : Consulting and training center - KEY, 2020)

TY  - CONF
AU  - Gazikalović, Ivana
AU  - Jovanović, Jelena
AU  - Šekuljica, Nataša
AU  - Luković, Nevena
AU  - Jakovetić Tanasković, Sonja
AU  - Knežević-Jugović, Zorica
PY  - 2020
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/6254
AB  - In modern times wheat gluten has drawn attention to many research groups.
Wheat gluten represents one of the most widely used proteins in the food
industry. It is a byproduct of the starch industry and has a higher percentage
of protein content compared to other plant-based protein sources. In order to
help reduce the allergenicity of wheat gluten, bacterial enzymes have been
proven to have beneficial results in wheat gluten treatment. In search for
an extracellular peptidase producing strain we have tested Bacillus subtilis
TMF-1 isolate, which has previously been proven to have several enzyme
activities. B. subtilis TMF-1 isolate has a food grade status, making it safe
for application in the food industry. Thus, the aim of this research was to
examine the possibility of utilizing mentioned strain in terms of glutendegrading
enzyme production.
Tested strain was first streaked onto several agar plates in order to detect
extracellular peptidase activity. Bacterial isolate has then been sequentially
transferred to the same growth medium several times. Conditions varied for
the submerged fermentation in 25 mL flasks were pH value of fermentation
broth, concentration of gluten powder (0 - 10 g/L) in fermentation broth
and concentration of peptone (0 - 1 g/L). Shaking flasks containing the
fermentation broth with the bacterial strain were kept for 48 h at 37 0C.
The results obtained show that the isolate has the possibility of thriving
in low acidic to neutral pH values of the fermentation broth. Varied gluten concentrations showed that even 1 g/L of gluten powder was sufficient for the
bacterial strain to manifest extracellular proteolytic enzyme activity. Peptone
concentrations were also varied, but even the minimal presence of peptone
has proven beneficial for bacterial growth and proteolytic activity.
This research show that the B. subtilis TMF-1 isolate has proteolytic activity
specific for wheat gluten as substrate and that it may be used in further
research in order to utilize its enzymatic production abilities for lowering
wheat gluten allergenicity.
PB  - Skopje : Consulting and training center - KEY
C3  - Book of abstracts / Nutricon 2020 - Food Quality and Safety, Health and Nutrition Congress
T1  - Optimization of submerged fermentation conditions for gluten-degrading enzyme production using B. subtilis isolate
EP  - 86
SP  - 85
UR  - https://hdl.handle.net/21.15107/rcub_technorep_6254
ER  - 
@conference{
author = "Gazikalović, Ivana and Jovanović, Jelena and Šekuljica, Nataša and Luković, Nevena and Jakovetić Tanasković, Sonja and Knežević-Jugović, Zorica",
year = "2020",
abstract = "In modern times wheat gluten has drawn attention to many research groups.
Wheat gluten represents one of the most widely used proteins in the food
industry. It is a byproduct of the starch industry and has a higher percentage
of protein content compared to other plant-based protein sources. In order to
help reduce the allergenicity of wheat gluten, bacterial enzymes have been
proven to have beneficial results in wheat gluten treatment. In search for
an extracellular peptidase producing strain we have tested Bacillus subtilis
TMF-1 isolate, which has previously been proven to have several enzyme
activities. B. subtilis TMF-1 isolate has a food grade status, making it safe
for application in the food industry. Thus, the aim of this research was to
examine the possibility of utilizing mentioned strain in terms of glutendegrading
enzyme production.
Tested strain was first streaked onto several agar plates in order to detect
extracellular peptidase activity. Bacterial isolate has then been sequentially
transferred to the same growth medium several times. Conditions varied for
the submerged fermentation in 25 mL flasks were pH value of fermentation
broth, concentration of gluten powder (0 - 10 g/L) in fermentation broth
and concentration of peptone (0 - 1 g/L). Shaking flasks containing the
fermentation broth with the bacterial strain were kept for 48 h at 37 0C.
The results obtained show that the isolate has the possibility of thriving
in low acidic to neutral pH values of the fermentation broth. Varied gluten concentrations showed that even 1 g/L of gluten powder was sufficient for the
bacterial strain to manifest extracellular proteolytic enzyme activity. Peptone
concentrations were also varied, but even the minimal presence of peptone
has proven beneficial for bacterial growth and proteolytic activity.
This research show that the B. subtilis TMF-1 isolate has proteolytic activity
specific for wheat gluten as substrate and that it may be used in further
research in order to utilize its enzymatic production abilities for lowering
wheat gluten allergenicity.",
publisher = "Skopje : Consulting and training center - KEY",
journal = "Book of abstracts / Nutricon 2020 - Food Quality and Safety, Health and Nutrition Congress",
title = "Optimization of submerged fermentation conditions for gluten-degrading enzyme production using B. subtilis isolate",
pages = "86-85",
url = "https://hdl.handle.net/21.15107/rcub_technorep_6254"
}
Gazikalović, I., Jovanović, J., Šekuljica, N., Luković, N., Jakovetić Tanasković, S.,& Knežević-Jugović, Z.. (2020). Optimization of submerged fermentation conditions for gluten-degrading enzyme production using B. subtilis isolate. in Book of abstracts / Nutricon 2020 - Food Quality and Safety, Health and Nutrition Congress
Skopje : Consulting and training center - KEY., 85-86.
https://hdl.handle.net/21.15107/rcub_technorep_6254
Gazikalović I, Jovanović J, Šekuljica N, Luković N, Jakovetić Tanasković S, Knežević-Jugović Z. Optimization of submerged fermentation conditions for gluten-degrading enzyme production using B. subtilis isolate. in Book of abstracts / Nutricon 2020 - Food Quality and Safety, Health and Nutrition Congress. 2020;:85-86.
https://hdl.handle.net/21.15107/rcub_technorep_6254 .
Gazikalović, Ivana, Jovanović, Jelena, Šekuljica, Nataša, Luković, Nevena, Jakovetić Tanasković, Sonja, Knežević-Jugović, Zorica, "Optimization of submerged fermentation conditions for gluten-degrading enzyme production using B. subtilis isolate" in Book of abstracts / Nutricon 2020 - Food Quality and Safety, Health and Nutrition Congress (2020):85-86,
https://hdl.handle.net/21.15107/rcub_technorep_6254 .

Modification of emulsifying properties and metal-ion chelating ability of gluten hydrolysates by partial enzymatic hydrolysis

Jovanović, Jelena; Stefanović, Andrea; Šekuljica, Nataša; Gazikalović, Ivana; Luković, Nevena; Jakovetić Tanasković, Sonja; Knežević-Jugović, Zorica

(Bratislava, SK : Slovak Society of Chemical Engineering, 2019)

TY  - CONF
AU  - Jovanović, Jelena
AU  - Stefanović, Andrea
AU  - Šekuljica, Nataša
AU  - Gazikalović, Ivana
AU  - Luković, Nevena
AU  - Jakovetić Tanasković, Sonja
AU  - Knežević-Jugović, Zorica
PY  - 2019
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/6260
AB  - Wheat gluten represents the major protein fraction present in wheat flour and as protein-rich
material posses cohesive and viscoelastic properties which permit the retention of gas bubbles in the dough. It could be obtained as a by-product during the separation of starch from wheat flour and as such may be utilized like a functional protein additive in various non-bakery foodstuffs due to its desirable structure-enhancing properties. However, wheat gluten is hardly soluble in water, which limits their expanding utilization. In the regard, the aim of this research was to investigate the correlation between process parameters of wheat gluten hydrolysis and emulsification activity and stability of the prepared hydrolysates. The hydrolysates showing the greatest enhancement of the emulsifying properties which are closed correlated with solubility improvement was further separated by sequential ultrafiltration to obtain molecular weight distribution profile and peptide fraction with higher metal-ion chelating ability. The hydrolysis was performed by using commercial endopeptidase from Bacillus licheniformis and process was followed by monitoring the degree of hydrolysis, emulsifying properties and metal-ion chelating activity. The effects of some relevant process parameters such as gluten concentration (1-9% w/v), temperature (40-60 °C), pH (7-9), enzyme/substrate ratio (0.25-0.75 AU g-¹ of protein) and their interactions were investigated by the means of the four-factor Box-Behnken experimental design with 29 experimental points (5 central points). The obtained results showed that the second-order models developed for emulsification activity and stability of gluten hydrolysates were significant (p<0.05) with a high value of the coefficients of determination (0.965-0.995). The statistical analysis showed that each variable had a significant effect on the emulsifying properties of the tested system. In terms of
emulsifying properties results showed that gluten concentration and temperature have had a positive effect on the increase of emulsification activity, while the enhancement of emulsification stability was achieved with the highest gluten concentration and pH 9. It appeared that the hydrolysate with great emulsifying properties had the highest percentage of peptides with medium molecular weight (3-10 kDa) which had the ability to strongly chelating prooxidant metal ions such as Fe2+ at level 99.3%. Results are substantial because they give useful information for the design an efficient process of gluten hydrolysis for production in high peptide yields with improved emulsifying properties. Also, may be suggesting that there are peptides with considerable size presenting a remarkable metal-ion chelating ability.
PB  - Bratislava, SK : Slovak Society of Chemical Engineering
C3  - PROCEEDINGS / 46th International Conference of the Slovak Society of Chemical Engineering
T1  - Modification of emulsifying properties and metal-ion chelating ability of gluten hydrolysates by partial enzymatic hydrolysis
EP  - 101-14
SP  - 101-1
UR  - https://hdl.handle.net/21.15107/rcub_technorep_6260
ER  - 
@conference{
author = "Jovanović, Jelena and Stefanović, Andrea and Šekuljica, Nataša and Gazikalović, Ivana and Luković, Nevena and Jakovetić Tanasković, Sonja and Knežević-Jugović, Zorica",
year = "2019",
abstract = "Wheat gluten represents the major protein fraction present in wheat flour and as protein-rich
material posses cohesive and viscoelastic properties which permit the retention of gas bubbles in the dough. It could be obtained as a by-product during the separation of starch from wheat flour and as such may be utilized like a functional protein additive in various non-bakery foodstuffs due to its desirable structure-enhancing properties. However, wheat gluten is hardly soluble in water, which limits their expanding utilization. In the regard, the aim of this research was to investigate the correlation between process parameters of wheat gluten hydrolysis and emulsification activity and stability of the prepared hydrolysates. The hydrolysates showing the greatest enhancement of the emulsifying properties which are closed correlated with solubility improvement was further separated by sequential ultrafiltration to obtain molecular weight distribution profile and peptide fraction with higher metal-ion chelating ability. The hydrolysis was performed by using commercial endopeptidase from Bacillus licheniformis and process was followed by monitoring the degree of hydrolysis, emulsifying properties and metal-ion chelating activity. The effects of some relevant process parameters such as gluten concentration (1-9% w/v), temperature (40-60 °C), pH (7-9), enzyme/substrate ratio (0.25-0.75 AU g-¹ of protein) and their interactions were investigated by the means of the four-factor Box-Behnken experimental design with 29 experimental points (5 central points). The obtained results showed that the second-order models developed for emulsification activity and stability of gluten hydrolysates were significant (p<0.05) with a high value of the coefficients of determination (0.965-0.995). The statistical analysis showed that each variable had a significant effect on the emulsifying properties of the tested system. In terms of
emulsifying properties results showed that gluten concentration and temperature have had a positive effect on the increase of emulsification activity, while the enhancement of emulsification stability was achieved with the highest gluten concentration and pH 9. It appeared that the hydrolysate with great emulsifying properties had the highest percentage of peptides with medium molecular weight (3-10 kDa) which had the ability to strongly chelating prooxidant metal ions such as Fe2+ at level 99.3%. Results are substantial because they give useful information for the design an efficient process of gluten hydrolysis for production in high peptide yields with improved emulsifying properties. Also, may be suggesting that there are peptides with considerable size presenting a remarkable metal-ion chelating ability.",
publisher = "Bratislava, SK : Slovak Society of Chemical Engineering",
journal = "PROCEEDINGS / 46th International Conference of the Slovak Society of Chemical Engineering",
title = "Modification of emulsifying properties and metal-ion chelating ability of gluten hydrolysates by partial enzymatic hydrolysis",
pages = "101-14-101-1",
url = "https://hdl.handle.net/21.15107/rcub_technorep_6260"
}
Jovanović, J., Stefanović, A., Šekuljica, N., Gazikalović, I., Luković, N., Jakovetić Tanasković, S.,& Knežević-Jugović, Z.. (2019). Modification of emulsifying properties and metal-ion chelating ability of gluten hydrolysates by partial enzymatic hydrolysis. in PROCEEDINGS / 46th International Conference of the Slovak Society of Chemical Engineering
Bratislava, SK : Slovak Society of Chemical Engineering., 101-1-101-14.
https://hdl.handle.net/21.15107/rcub_technorep_6260
Jovanović J, Stefanović A, Šekuljica N, Gazikalović I, Luković N, Jakovetić Tanasković S, Knežević-Jugović Z. Modification of emulsifying properties and metal-ion chelating ability of gluten hydrolysates by partial enzymatic hydrolysis. in PROCEEDINGS / 46th International Conference of the Slovak Society of Chemical Engineering. 2019;:101-1-101-14.
https://hdl.handle.net/21.15107/rcub_technorep_6260 .
Jovanović, Jelena, Stefanović, Andrea, Šekuljica, Nataša, Gazikalović, Ivana, Luković, Nevena, Jakovetić Tanasković, Sonja, Knežević-Jugović, Zorica, "Modification of emulsifying properties and metal-ion chelating ability of gluten hydrolysates by partial enzymatic hydrolysis" in PROCEEDINGS / 46th International Conference of the Slovak Society of Chemical Engineering (2019):101-1-101-14,
https://hdl.handle.net/21.15107/rcub_technorep_6260 .