Secondary structure by circular dichroism spectroscopy of β-N-acetylglucosaminidase from Lecanicillium lecanii and its relationship with hydrolytic and transglycosylation activities at different pH values

Abstract

N-acetylglucosaminidase produced and purified from Lecanicillium lecanii was analyzed using circular dichroism spectroscopy. The enzyme is mainly an (α + β)-type protein, having 76 % α-helix, 4 % parallel β-sheet, 2 % antiparallel β-sheet, 8 % β-turns, and 10 % unordered structure at pH 10 (condition leading to the highest secondary structure content, 100% ellipticity). The circular dichroism spectra analysis as a function of pH shows a decrease in α-helix content at pH < 8. The structure-activity relationship as a function of pH to hydrolytic or transglycosylation reactions was studied. The maximum transglycosylation activity for four donor substrates was at pH 7, while hydrolytic activity was at pH 6. The highest activity pH values do not correspond to the maximum percentage of ellipticity for both. The maximum hydrolytic or transglycosylation activity observed is associated with conformational changes in the enzyme when pH diminishes.

Keywords: N-acetylglucosaminidase, Lecanicillium lecanii, circular dichroism, hydrolytic, transglycosylation.

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