Diol oxidation in the presence of oxidoreductases

U ovom radu istražena je enzimatska oksidacija alkohola katalizirana enzimom galaktoza oksidazom. Zanimala nas je specifična aktivnost galaktoza oksidaze prema alkoholnom supstratu, diolu. Provedeno je mjerenje utjecaja koncentracije diola, bakrovih iona i kisika na aktivnost enzima. Kao supstrati su korišteni racemična smjesa diola, ali i čisti R- i S- izomeri diola, kako bi se pokazalo je li enzim enantioselektivan. Postojanje produkta enzimatske reakcije oksidacije alkohola ispitano je kapljevinskom kromatografijom visoke djelotvornosti i kombinirane tehnike kapljevinske kromatografije i masene spektrometrije. Provedena je i kaskadna reakcija u kojoj je oksidacija alkohola kombinirana s aldolnom adicijom uz enzim D-fruktoza-6-fosfat aldolazu (FSA).In this work, enzymatic alcohol oxidation catalyzed by galactose oxidase was studied. We were interested in the specific activity of galactose oxidase to the alcohol substrate. Measurement of the effect of diol, copper ions and oxygen concentration on enzyme activity was performed. The racemic alcohol was tested as substrate, but also the R- and S-isomers. The existence of enzymatic reaction products was tested by high-performance liquid chromatography and combined liquid chromatography and mass spectrometry techniques. Cascade reactions of alcohol oxidation and aldol addition to D-fructose-6-phosphate aldolase (FSA) were also performed