Characterization of β-galactosidase from Lactobacillus acidophilus: Stability and kinetic study
Produkcija i karakterizacija β-galaktozidaze iz Lactobacillus acidophilus
Samo za registrovane korisnike
2017
Autori
Simović, MilicaVukašinović-Sekulić, Maja
Banjanac, Katarina
Milivojević, Ana
Ćorović, Marija
Bezbradica, Dejan
Članak u časopisu (Objavljena verzija)
Metapodaci
Prikaz svih podataka o dokumentuApstrakt
β-Galactosidase is the industrially important enzyme that catalyzes both, lactose hydrolysis and synthesis of different bioactive galactosides. In this study, optimal conditions (fermentation temperature, inoculum concentration and lactose concentration) for accomplishing high yields of β-galactosidase activity from Lactobacillus acidophilus ATCC 4356, bacteria regarded as safe for human consumption, were investigated. Using the response surface methodology (RSM), it was concluded that the highest activity and specific activity were obtained by 2-day shake-flask culture fermentation at 28°C, provided that the lactose content in the fermentation medium was 1.48%, and the inoculum concentration was 2.8%. The optimum temperature and pH for the obtained enzyme were 45°C and 6.5, respectively. More importantly, these conditions simultaneously ensure a high enzyme stability. The Km and Vmax values were 0.44 mM and 25.64 mM/h (for o-nitrophenyl-β-D-galactopyranoside), and 3.79 mM and 3.10 mM/...h (for lactose), and the substrate excess inhibition was not observed. On the other hand, the enzyme was inactivated in the presence of Ca2+, Ba2+, and Cu2+.
β-Galaktozidaza je značajan industrijski enzim koji se tradicionalno koristi za katalizovanje reakcije hidroloze laktoze, ali poslednjih godina nalazi i sve značajniju primenu u reakcijama sinteze različitih bioaktivnih galaktozida. U ovoj studiji, ispitani su optimalni uslovi za postizanje visokih prinosa β-galaktozidaze iz Lactobacillus acidophilus ATCC 4356, bakterije mlečne kiseline koja se smatra bezbednom za korišćenje u prehrambenim proizvodima. Statističkim planiranjem eksperimenata i metodom odzivnih površina, zaključeno je da se najveća aktivnost i specifična aktivnost dobijaju nakon dvodnevne fermentacije na temperaturi od 28°S, kada je koncentracija laktoze 1,48%, a koncentracija inokuluma 2,80%. Temperaturni i rN optimum dobijenog enzima iznose 45°S i 6,8, redom, pri čemu ovi uslovi istovremeno omogućuju i izuzetnu stabilnost enzima. U kinetičkoj studiji određene su vrednosti kinetičkih parametara, Km i Vmax: 0,44 mM i 25,64 mM/h (za o-nitrofenil- β-D-galaktopiranozid), od...nosno 3,79 mM i 3,10 mM/h (za laktozu). Takođe, nije primećena inhibicija supstratom u višku, dok se enzim inaktivira u prisustvu jona Ca2+, Ba2+, i Cu2+. PR Projekat Ministarstva nauke Republike Srbije, br. III 46010 .
Ključne reči:
β-galactosidase / Lactobacillus acidophilus / enzyme production / response surface methodology (RSM) / β-galaktozidaza / Lactobacillus acidophilus / proizvodnja enzima / metoda odzivnih površinaIzvor:
Advanced Technologies, 2017, 6, 1, 5-13Izdavač:
- Univerzitet u Nišu - Tehnološki fakultet, Leskovac
Finansiranje / projekti:
- Razvoj novih inkapsulacionih i enzimskih tehnologija za proizvodnju biokatalizatora i biološki aktivnih komponenata hrane u cilju povećanja njene konkurentnosti, kvaliteta i bezbednosti (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-46010)
Kolekcije
Institucija/grupa
Tehnološko-metalurški fakultetTY - JOUR AU - Simović, Milica AU - Vukašinović-Sekulić, Maja AU - Banjanac, Katarina AU - Milivojević, Ana AU - Ćorović, Marija AU - Bezbradica, Dejan PY - 2017 UR - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3505 AB - β-Galactosidase is the industrially important enzyme that catalyzes both, lactose hydrolysis and synthesis of different bioactive galactosides. In this study, optimal conditions (fermentation temperature, inoculum concentration and lactose concentration) for accomplishing high yields of β-galactosidase activity from Lactobacillus acidophilus ATCC 4356, bacteria regarded as safe for human consumption, were investigated. Using the response surface methodology (RSM), it was concluded that the highest activity and specific activity were obtained by 2-day shake-flask culture fermentation at 28°C, provided that the lactose content in the fermentation medium was 1.48%, and the inoculum concentration was 2.8%. The optimum temperature and pH for the obtained enzyme were 45°C and 6.5, respectively. More importantly, these conditions simultaneously ensure a high enzyme stability. The Km and Vmax values were 0.44 mM and 25.64 mM/h (for o-nitrophenyl-β-D-galactopyranoside), and 3.79 mM and 3.10 mM/h (for lactose), and the substrate excess inhibition was not observed. On the other hand, the enzyme was inactivated in the presence of Ca2+, Ba2+, and Cu2+. AB - β-Galaktozidaza je značajan industrijski enzim koji se tradicionalno koristi za katalizovanje reakcije hidroloze laktoze, ali poslednjih godina nalazi i sve značajniju primenu u reakcijama sinteze različitih bioaktivnih galaktozida. U ovoj studiji, ispitani su optimalni uslovi za postizanje visokih prinosa β-galaktozidaze iz Lactobacillus acidophilus ATCC 4356, bakterije mlečne kiseline koja se smatra bezbednom za korišćenje u prehrambenim proizvodima. Statističkim planiranjem eksperimenata i metodom odzivnih površina, zaključeno je da se najveća aktivnost i specifična aktivnost dobijaju nakon dvodnevne fermentacije na temperaturi od 28°S, kada je koncentracija laktoze 1,48%, a koncentracija inokuluma 2,80%. Temperaturni i rN optimum dobijenog enzima iznose 45°S i 6,8, redom, pri čemu ovi uslovi istovremeno omogućuju i izuzetnu stabilnost enzima. U kinetičkoj studiji određene su vrednosti kinetičkih parametara, Km i Vmax: 0,44 mM i 25,64 mM/h (za o-nitrofenil- β-D-galaktopiranozid), odnosno 3,79 mM i 3,10 mM/h (za laktozu). Takođe, nije primećena inhibicija supstratom u višku, dok se enzim inaktivira u prisustvu jona Ca2+, Ba2+, i Cu2+. PR Projekat Ministarstva nauke Republike Srbije, br. III 46010 . PB - Univerzitet u Nišu - Tehnološki fakultet, Leskovac T2 - Advanced Technologies T1 - Characterization of β-galactosidase from Lactobacillus acidophilus: Stability and kinetic study T1 - Produkcija i karakterizacija β-galaktozidaze iz Lactobacillus acidophilus EP - 13 IS - 1 SP - 5 VL - 6 DO - 10.5937/savteh1701005C ER -
@article{ author = "Simović, Milica and Vukašinović-Sekulić, Maja and Banjanac, Katarina and Milivojević, Ana and Ćorović, Marija and Bezbradica, Dejan", year = "2017", abstract = "β-Galactosidase is the industrially important enzyme that catalyzes both, lactose hydrolysis and synthesis of different bioactive galactosides. In this study, optimal conditions (fermentation temperature, inoculum concentration and lactose concentration) for accomplishing high yields of β-galactosidase activity from Lactobacillus acidophilus ATCC 4356, bacteria regarded as safe for human consumption, were investigated. Using the response surface methodology (RSM), it was concluded that the highest activity and specific activity were obtained by 2-day shake-flask culture fermentation at 28°C, provided that the lactose content in the fermentation medium was 1.48%, and the inoculum concentration was 2.8%. The optimum temperature and pH for the obtained enzyme were 45°C and 6.5, respectively. More importantly, these conditions simultaneously ensure a high enzyme stability. The Km and Vmax values were 0.44 mM and 25.64 mM/h (for o-nitrophenyl-β-D-galactopyranoside), and 3.79 mM and 3.10 mM/h (for lactose), and the substrate excess inhibition was not observed. On the other hand, the enzyme was inactivated in the presence of Ca2+, Ba2+, and Cu2+., β-Galaktozidaza je značajan industrijski enzim koji se tradicionalno koristi za katalizovanje reakcije hidroloze laktoze, ali poslednjih godina nalazi i sve značajniju primenu u reakcijama sinteze različitih bioaktivnih galaktozida. U ovoj studiji, ispitani su optimalni uslovi za postizanje visokih prinosa β-galaktozidaze iz Lactobacillus acidophilus ATCC 4356, bakterije mlečne kiseline koja se smatra bezbednom za korišćenje u prehrambenim proizvodima. Statističkim planiranjem eksperimenata i metodom odzivnih površina, zaključeno je da se najveća aktivnost i specifična aktivnost dobijaju nakon dvodnevne fermentacije na temperaturi od 28°S, kada je koncentracija laktoze 1,48%, a koncentracija inokuluma 2,80%. Temperaturni i rN optimum dobijenog enzima iznose 45°S i 6,8, redom, pri čemu ovi uslovi istovremeno omogućuju i izuzetnu stabilnost enzima. U kinetičkoj studiji određene su vrednosti kinetičkih parametara, Km i Vmax: 0,44 mM i 25,64 mM/h (za o-nitrofenil- β-D-galaktopiranozid), odnosno 3,79 mM i 3,10 mM/h (za laktozu). Takođe, nije primećena inhibicija supstratom u višku, dok se enzim inaktivira u prisustvu jona Ca2+, Ba2+, i Cu2+. PR Projekat Ministarstva nauke Republike Srbije, br. III 46010 .", publisher = "Univerzitet u Nišu - Tehnološki fakultet, Leskovac", journal = "Advanced Technologies", title = "Characterization of β-galactosidase from Lactobacillus acidophilus: Stability and kinetic study, Produkcija i karakterizacija β-galaktozidaze iz Lactobacillus acidophilus", pages = "13-5", number = "1", volume = "6", doi = "10.5937/savteh1701005C" }
Simović, M., Vukašinović-Sekulić, M., Banjanac, K., Milivojević, A., Ćorović, M.,& Bezbradica, D.. (2017). Characterization of β-galactosidase from Lactobacillus acidophilus: Stability and kinetic study. in Advanced Technologies Univerzitet u Nišu - Tehnološki fakultet, Leskovac., 6(1), 5-13. https://doi.org/10.5937/savteh1701005C
Simović M, Vukašinović-Sekulić M, Banjanac K, Milivojević A, Ćorović M, Bezbradica D. Characterization of β-galactosidase from Lactobacillus acidophilus: Stability and kinetic study. in Advanced Technologies. 2017;6(1):5-13. doi:10.5937/savteh1701005C .
Simović, Milica, Vukašinović-Sekulić, Maja, Banjanac, Katarina, Milivojević, Ana, Ćorović, Marija, Bezbradica, Dejan, "Characterization of β-galactosidase from Lactobacillus acidophilus: Stability and kinetic study" in Advanced Technologies, 6, no. 1 (2017):5-13, https://doi.org/10.5937/savteh1701005C . .