Chemical and enzymatic oxidation of Cbz-N-3-amino-1,2-propanediol

U ovom radu je provedena kemijska i enzimatska oksidacija aminoalkohola. Kemijska oksidacija Cbz-N-3-amino-1,2-propandiola provedena je uz prisustvo jodoksibenzojeve kiseline, a enzimatska uz alkohol dehidrogenazu. Alkohol dehidrogenaza za svoje katalitičko djelovanje zahtjeva koenzim NAD+. Kako ga ne bi bilo potrebno dodavati u reaktor u stehiometrijskim količinama, regeneriran je uz korištenje NADH oksidaze iz Lactococcus lactis, koja je za potrebe ovog rada izolirana i pročišćena. Određena je kinetika reakcije oksidacije aminoalkohola katalizirana s alkohol dehidrogenazom te su procijenjeni kinetički parametri. Ispitan je utjecaj omjera količina enzima alkohol dehidrogenaze i NADH oksidaze na ishod oksidacije. Najpovoljniji omjer je korišten u kaskadnoj reakciji oksidacije aminoalkohola i slijedne reakcije aldolne adicije u kojoj je sintetiziran aldolni produkt.Chemical and enzymatic oxidation of the amino alcohol was carried out in this work. Chemical oxidation of Cbz-N-3-amino-1,2-propanediol was carried out by iodoxybenzoic acid and enzymatic oxidation by alcohol dehydrogenase. Alcohol dehydrogenase requires coenzyme NAD+ for its catalytic activity. Since it would be too expensive to add it in the reactor in stoichiometric amounts, it was regenerated by using NADH oxidase from Lactococcus lactis. This enzyme was isolated and purified for the purpose of this work. Kinetics of the amino alcohol oxidation catalyzed by alcohol dehydrogenase was determined and the kinetic parameters were estimated. The influence of the ratio of the amount of alcohol dehydrogenase and NADH oxidase on the outcome of the oxidation was examined. The most appropriate ratio was used in the cascade oxidation of the amino alcohol and sequential reaction of aldol addition in which aldol adduct was formed