| The biocatalysis of a crude chlorophyllase extract, obtained from the biomass culture of Phaeodactylum tricornutum, in neat organic solvent media was investigated. The addition of selected excipients, including crown ether (enzyme:crown ether, 135:1--2.7:1, w/w), dextran (enzyme:dextran, 1:2--1:0.25, w/w), Span 40 and Span 60 (enzyme:Span, 1:2, w/w) and sodium bis (2-ethylhexyl) sulfosuccinate (enzyme:AOT, 1:12.6, w/w), to the crude solid enzyme preparation decreased the chlorophyllase activity. The effects of selected parameters, including solvent hydrophobicity (Log P, 2.40--4.45), initial water activity (aw, 0.44--0.97), agitation speed (0--200 rpm), reaction temperature (25--45°C) and enzyme concentration (1.67--5.3 mg solid enzyme/mL) on chlorophyllase activity, were investigated using a crude solid enzyme. The experimental findings showed that the highest chlorophyllase specific activity of 362.4 nmol hydrolyzed chlorophyll/g solid enzyme/min and bioconversion yield of 90.7% were obtained with the hexane/octanone mixture (98.7:1.3, v/v), aw of 0.90, agitation speed of 200 rpm, reaction temperature of 35°C and enzyme concentration of 3.33 mg solid enzyme/mL. (Abstract shortened by UMI.)... |
