DOI - Mendel University Press

DOI identifiers

DOI: 10.11118/978-80-7509-994-5-0022

MELANIN IN BARLEY: FROM ISOLATION TO A POSSIBILITY TO INFLUENCE THE ACTIVITY OF BIOTRANSFORMATION ENZYMES

V. BATKOVÁ1, L. JOUROVÁ1, Š. ŠATKA1, V. FRÝBORTOVÁ1, P. ANZENBACHER2, E. MRKVICOVÁ3, P. MARTÍNEK4, E. ANZENBACHEROVÁ1
1 Department of Medical Chemistry and Biochemistry Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic
2 Department of Pharmacology, Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic
3 Department of Animal Nutrition and Forage Production, Faculty of AgriSciences, Mendel University in Brno, Zemědělská 1, 613 00, Brno, Czech Republic
4 Agrotest fyto, Ltd., 76701 Kroměříž, Czech Republic

This study describes the isolation and characterization of melanin from plant matrices using a modified version of the alkaline extraction method originally described by Sava et al. (2001). Melanin was isolated from barley (variety Nudimelanocriton) and purified through a series of organic solvent treatments, acidic hydrolysis, and repeated precipitations, yielding approximately 5 mg of pure melanin from 12 g of barley grain. Isolated melanin exhibited its characteristic properties as insolubility in water, acids, and organic solvents, while being soluble in alkaline media and precipitable below pH 3. Consistent with findings from Caldas et al. (2020) and Glagoleva et Shoeva (2020), melanin samples exhibited in alkaline media a broad-band almost monotonous decrease of UV/VIS absorption from initially 200 nm, with unresolved absorption band at about 270 nm, indicative of complex conjugated structures of aromatic character. Additionally, effect of melanin on cytochrome P450 1A1/2 enzyme activity was assessed in HepG2 cells using 7-ethoxyresorufin O-dealkylation and high-performance liquid chromatography. Melanin at three concentrations (10 μg/mL, 1 μg/mL and 0.1 μg/mL) did not significantly induce cytochrome P450 1A1/2 enzyme activity (in contrast to CYP1A1/2 potent inducer, 2,3,7,8-tetrachlordibenzodioxin resulting in a nearly sixtyfold increase). These findings contribute to understanding the physicochemical properties of barley-derived melanin and its interaction with hepatic enzymes of xenobiotic biotransformation (as CYP1A1/2).

Keywords: melanin; allomelanin; barley; cytochrome P450; enzyme activity; cell culture; HepG2; UV-visible spectrophotometry

pages: 22-36, online: 2024



Correction:

On 9th September, page 35 was corrected. The last number was omitted from the number of project NAZV. The correct number is NAZV QL24010230.

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