Yong-Joon Yang

Production of protocatechualdehyde using tyrosinase purified from potato

Tyrosinase is responsible for the enzymatic browning of fruits and vegetables and usually catalyze the conversion of monophenols to o-diphenols and oxidation of diphenols to corresponding quinines. However, when 3,4-dihydroxymandelic acid is provided as the substrate, it catalyzes the oxidative decarboxylation reaction to generate 3,4-dihydroxybenzaldehyde (protocatechualdehyde, PA. In the present study, tyrosinase was purified from potato with 27.9 purification fold by salting out with ammonium sulfate, DEAE-Sepharose and Sephadex G-150 column chromatography. The purified tyrosinase was confirmed by tyrosinase active staining following SDS-PAGE. Tyrosinase activity was visualized in the gel as a dark band. The molecular weight of tyrosinase from potato was 38kDa as determined by SDS-PAGE. Purified tyrosinase mediated oxidative decarboxylation of 3,4-dihydroxymandelate. The identity of the reaction product, PA was confirmed by high-performance liquid chromatography (HPLC) as well as ultraviolet spectral studies. Phenol oxidase inhibitors such as potassium cyanide, sodium azide, and phenylthiourea inhibited the participation of the active site copper of the enzyme in the catalysis.

Tyrosinase inhibitory activity and antimicrobial activity by mixtures of ultrasonicated chitosan and Maillard reaction products

The aim of this study was to investigate the tyrosinase inhibitory activity and antimicrobial activity by mixtures of ultrasonicated chitosan and Maillard reaction products. Analysis of tyrosinase was purified from potato and confirmed by active staining after SDS-PAGE. Maillard reaction products (MRPs) were formed from various sugars (glucose, fructose, galactose, xylose, arabinose or ribose) and cystein. MRPs inhibited the tyrosinase purified from potato. The highest tyrosinase inhibitory activity was shown by MRP from glucose and cystein. Ultrasonicated chitosan (over 1 hr) showed antimicrobial activity at the concentration of 1% against E. coli and S. aureus. For the development of antibrowning agent with antimicrobial activity, tyrosinase inhibitory and antimicrobial activity by the mixtures of ultrasonicated chitosan and MRP were tested. 1:1 mixture of ultrasonicated chitosan and MRP from glucose and cystein was the best antibrowning agent having antimicrobial activity.