This work established an integrated utilization of dairy whey in β-galactosidase production from Lactobacillus bulgaricus and prebiotics synthesis by the probiotic enzyme. A cost-effective whey-based medium was newly developed for culturing Lactobacillus bulgaricus to produce β-galactosidase. The medium was optimized through response surface methodology (RSM) involving a series of statistical designs, such as the Plackett–Burman design, steepest ascent experiment, and central composite design. Under the optimized medium, the β-galactosidase activity of L. bulgaricus reached 2034 U/L, which was twice that produced from the traditional MRS medium. The cells of L. bulgaricus harvested from the whey-based medium were subsequently treated with lysozyme. The resulting crude enzyme was used as an efficient catalyst, which catalyzed the synthesis of the prebiotic galacto-oligosaccharides (GOS) in a high yield of 44.7% by using whey (200 g/L) as the substrate. The sugar mixture was further purified by activated charcoal adsorption, thereby yielding a high-purity level of 77.6% GOS.
Herein, ethanol], which is rich in olive oil and red wine, was converted to a novel bioactive galactoside by enzymic glycosylation. The gene of α-galactosidase from Geobacillus stearothermophilus 23 was cloned and expressed in Escherichia coli as catalytically active inclusion bodies. The catalytically active inclusion bodies efficiently catalyzed the galactosylation of tyrosol using either melibiose or raffinose family oligosaccharides as glycosyl donors, resulting in a glycoside with 42.2 or 14.2% yields. The glycoside product was purified and identified as p-hydroxyphenethyl α-D-galactopyranoside by mass spectrometry and NMR analyses. The inclusion bodies can be recycled and reused for at least 10 batch reactions of galactoside synthesis. Moreover, the galactoside showed 11-fold increased water solubility and reduced cytotoxicity as compared to tyrosol. Also, it exhibited higher antioxidative and anti-inflammatory activities than tyrosol based on lipopolysaccharide-induced activated BV2 cells. These results provided important insights into the application of tyrosol derivatives in functional foods.
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