Characterization of Β -galactosidase from Lactobacillus acidophilus : stability and kinetic study
2017
Carević, Milica B. (Faculty of Technology and Metallurgy, Belgrade (Serbia). Department of Biochemical Engineering and Biotechnology) | Vukašinović-Sekulić, Maja S. (Faculty of Technology and Metallurgy, Belgrade (Serbia). Department of Biochemical Engineering and Biotechnology) | Banjanac, Katarina M. (Faculty of Technology and Metallurgy, Belgrade (Serbia). Department of Biochemical Engineering and Biotechnology) | Milivojević, Ana D. (Faculty of Technology and Metallurgy, Belgrade (Serbia). Department of Biochemical Engineering and Biotechnology) | Ćorović, Marija M. (Faculty of Technology and Metallurgy, Belgrade (Serbia). Department of Biochemical Engineering and Biotechnology) | Bezbradica, Dejan I. (Faculty of Technology and Metallurgy, Belgrade (Serbia). Department of Biochemical Engineering and Biotechnology)
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 Ca 2+ , Ba 2+ , and Cu 2+ .
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