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Optimization of the process of biotransformation of hydroquinone into its β-D-glucoside – arbutin, was performed in agitated shoot cultures of Schisandra chinensis. The optimisation involved testing various concentrations of the precursor and different ways of administering it. Arbutin was accumulated mainly in the in vitro cultured biomass (85.2–98.6%). By optimizing the process, a 2.26-fold increase in the overall product content was obtained. The highest amount (17.8 mg·g–1 DW) was found after administering 384 mg·l–1 hydroquinone in a dose divided into two portions. An experiment with the biotransformation of 4-hydro- xybenzoic acid did not produce arbutin but a mixture of two products of glucosylation of the precursor – hydroxybenzoic acid 4-O-β-glucopyranoside and 4-hydroxybenzoic acid β-glucopyranosyl ester. The identity of all biotransformation products was confirmed by 1H-NMR analysis. The results for the production of arbutin by the biotransformation of hydroquinone are of potential practical importance. On the other hand, the fact of confirming the presence of two glucosylation products has a great cognitive value.
Arbutin (hydroquinone β-D-glucoside) is a compound of plant origin possessing valuable therapeutic (urinary tract disinfection) and cosmetic (skin whitening) properties, which can be obtained from in vitro cultures of plants belonging to different taxa via biotransformation of exogenously supplemented hydroquinone. Agitating cultures of Aronia melanocarpa were maintained on the Murashige and Skoog medium containing growth regulators: the cytokinin - BAP (6-benzylaminopurine), 2 mg/l and the auxin NAA (α-naphthaleneacetic acid), 2 mg/l. The biomass was cultured for 2 weeks and then hydroquinone was supplemented at the following doses: 96, 144, 192, 288 and 384 mg/l either undivided or divided into two or three portions added at 24-hour intervals. The content of the reaction product - arbutin, was determined using an HPLC method in methanolic extracts from biomass and lyophilized medium samples collected 24 hours after the addition of the last precursor dose. The total amounts of arbutin were very diverse, from 2.71 to 8.27 g/100g d.w. The production of arbutin rose with increasing hydroquinone concentration. The maximum content of the product was observed after hydroquinone addition at 384 mg/l divided into two portions. Biotransformation efficiency also varied widely, ranging from 37.04% do 73.80%. The identity of the product - arbutin, after its isolation and purification was confirmed by spectral analysis (1H-NMR spectrum). The maximum amount of arbutin obtained was higher than that required by the latest 9th Edition of the Polish Pharmacopoeia and by the newest 8th Edithion of European Pharmacopoeia for Uvae ursi folium (7.0 g/100g d.w.), and is interesting from practical point of view.
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