Enzymatic glycosylation is a versatile and sustainable biotechnological approach that plays a pivotal role in the production of bioactive compounds. This process involves the enzymatic transfer of sugar moieties onto various acceptor molecules, such as small molecules, peptides, or proteins, resulting in the synthesis of glycosides. These glycosides often exhibit enhanced bioactivity, improved solubility, and enhanced stability, making them valuable in pharmaceuticals, nutraceuticals, and the food industry. This review explores the diverse enzymatic glycosylation strategies employed in the synthesis of bioactive compounds. It highlights the enzymatic catalysts involved, including glycosyltransferases, glycosidases, glycophosphorylases, and glycosynthases. It considers the advantages and disadvantages of these biocatalysts in the stereoselective and regioselective synthesis of different types of glycosylated molecules, phenolic and aliphatic alcohols, oligosaccharides, polysaccharides, glycoderivatives, glycopeptides, and glycoproteins with a clear focus on food and pharmaceutical chemistry. Furthermore, the review outlines various sources of sugar donors, activated glycosides, and sugar nucleotides, as well as the utilization of engineered enzymes and microorganisms for glycosylation reactions. The advantages of enzymatic glycosylation, including its high regioselectivity, stereoselectivity, and sustainability, are emphasized. Therefore, these approaches combining the use of different catalytic systems, the improvement of tools such as immobilization technology or chemical or genetic modification to improve the glycosylation process, could be useful tools in continuous biotechnological advancements.

Background: Blackcurrant is a rich source of polyphenols with proven physiological activity, that have lately been classified as emerging prebiotics, making its extracts suitable for application in functional food and bioactive cosmetics. However, these biomolecules are trapped in cell wall structures and nowadays, enzymatic hydrolysis is recognised as powerful tool for extraction efficiency improvement in an environmentally sound manner. This study aimed to optimize enzyme-assisted extraction to maximize yields of blackcurrant polyphenols. In addition, their prebiotic potential was tested by examining their influence on typical skin microbiota representatives as model microorganisms—beneficial coagulase-negative Staphylococcus epidermidis and two opportunistic pathogens, Staphylococcus aureus and Cutibacterium acnes. Results: Among four examined commercial enzyme preparations, Viscozyme^® L and Rohapect^® MC, demonstrated the highest potential as extraction enhancers by increasing antioxidant activity, total polyphenol, and flavonoid contents. Furthermore, their synergetic effect enabled 95.3% increase in polyphenol concentration compared to conventional extraction with a maximum of 27.6 mg gallic acid equivalents (GAE)/g dry matter (DM) at a total enzyme concentration of 0.05 mL/g DM (Viscozyme^® L to Rohapect^® MC ratio 2:1) after 60 min. HPLC profiles of two samples showed differences in polyphenol composition indicating simultaneous extraction–biotransformation. At a concentration of 0.025 mg GAE/mL, PAS (prebiotic activity score) values were 0.096 and 0.172 for conventionally and enzymatically obtained extract, respectively. Moderate inhibitory effect of optimal extract concentration on Cutibacterium acnes was shown, as well. Hydrogel prepared with optimal extract concentration showed improved hydration effect, decreased irritability and increased dermatological compatibility comparing to basic formulation. Conclusions: Overall results demonstrated that blackcurrant extract obtained under optimized conditions using enzymes possesses an emerging prebiotic potential for selective skin microbiota stimulation. Graphical

Blackcurrant pomace is a widely available waste stream derived from the industrial production of juice rich in pectin and unextracted polyphenols. Since pectin, an emerging class of gastrointestinal prebiotics, is also a common cosmetic ingredient, the aim of this work was to evaluate blackcurrant pomace as a source of pectin-rich fractions suitable for application in prebiotic cosmetics. Hereby, this raw material was valorised by sequential extraction of acid-soluble (by citric acid, CAP) and Ca-bound (by ammonium oxalate, AOPP) pectic polysaccharides. Both fractions had favourable physicochemical features and a similar degree of methyl-esterification between low- and high-methoxyl pectin (approx. 50%), but CAP had significantly higher galacturonic acid content (72.3%), branching, and purity. Regardless of that, both had very high oil (18.96 mL/g for CAP and 19.32 mL/g for AOPP) and water (9.97 mL/g for CAP and 7.32 mL/g for AOPP)-holding capacities and excellent emulsifying properties, making them promising cosmetic ingredients. The polyphenol content was 10 times higher in CAP, while corresponding antioxidant activity was 3-fold higher. Finally, the influence of varying CAP and AOPP concentrations on common skin pathogen, Staphylococcus aureus, and beneficial skin bacteria, Staphylococcus epidermidis, was examined. The results show significant prebiotic potential of two pectic fractions since they were capable of selectively stimulating S. epidermidis, while S. aureus growth was inhibited, whereas CAP demonstrated a particularly high capacity of up to 2.2, even with methicillin-resistant S. aureus.

Blackcurrant is an anthocyanin-rich berry with proven antioxidant and photoprotective activity and emerging prebiotic potential, widely applied in cosmetic products. Hereby, highly efficient enzyme-assisted extraction of blackcurrant polyphenols was performed, giving extract with very high antioxidant activity. Obtained extract was characterized in terms of anthocyanin composition, incorporated into three different cosmetic formulations and subjected to Franz cell diffusion study. Experimental values obtained using cellulose acetate membrane for all four dominant anthocyanins (delphinidin 3-glucoside, delphinidin 3-rutinoside, cyanidin 3-glucoside and cyanidin 3-rutinoside) were successfully fitted with the Korsmeyer–Peppas diffusion model. Calculated effective diffusion coefficients were higher for hydrogel compared to oil-in-water cream gel and oil-in-water emulsion, whereas the highest value was determined for cyanidin 3-rutinoside. On the other hand, after a 72 h long experiment with transdermal skin diffusion model (Strat-M^® membrane), no anthocyanins were detected in the receptor fluid, and only 0.5% of the initial quantity from the donor compartment was extracted from the membrane itself after experiment with hydrogel. Present study revealed that hydrogel is a suitable carrier system for the topical delivery of blackcurrant anthocyanins, while dermal and transdermal delivery of these molecules is very limited, which implies its applicability for treatments targeting skin surface (i.e., prebiotic, photoprotective).