Izvorni znanstveni članak
Resolution of Racemic Acids, Esters and Amines by Candida rugosa Lipase in Slightly Hydrated Organic Media
Andrés R. Alcántara
; Biotransformations group, Department of Organic & Pharmaceutical Chemistry, Faculty of Pharmacy, Universidad Complutense, Plza. Ramón y Cajal s/n, Ciudad Universitaria, E-28040 Madrid, Spain
Pablo Domínguez de María
; Biotransformations group, Department of Organic & Pharmaceutical Chemistry, Faculty of Pharmacy, Universidad Complutense, Plza. Ramón y Cajal s/n, Ciudad Universitaria, E-28040 Madrid, Spain
María Fernández
; Biotransformations group, Department of Organic & Pharmaceutical Chemistry, Faculty of Pharmacy, Universidad Complutense, Plza. Ramón y Cajal s/n, Ciudad Universitaria, E-28040 Madrid, Spain
María José Hernaíz
; Biotransformations group, Department of Organic & Pharmaceutical Chemistry, Faculty of Pharmacy, Universidad Complutense, Plza. Ramón y Cajal s/n, Ciudad Universitaria, E-28040 Madrid, Spain
José María Sánchez-Montero
; Biotransformations group, Department of Organic & Pharmaceutical Chemistry, Faculty of Pharmacy, Universidad Complutense, Plza. Ramón y Cajal s/n, Ciudad Universitaria, E-28040 Madrid, Spain
José Vincente Sinisterra
; Biotransformations group, Department of Organic & Pharmaceutical Chemistry, Faculty of Pharmacy, Universidad Complutense, Plza. Ramón y Cajal s/n, Ciudad Universitaria, E-28040 Madrid, Spain
Sažetak
Commercial crude lipase from Candida rugosa is widely used as a biocatalyst in the resolution of racemic mixtures and in organic synthesis in slightly hydrated organic solvents. In many cases, reproducible results are not obtained when the same crude lipase is used, but from different suppliers of lots, this being due to the presence of different isoenzymes. The current work addresses this problem and strategies to overcome it. The yeast Candida rugosa ATCC 14380 was cultivated in a minimal culture medium, using different substances as inducers and carbon sources. The percentage of inducer that gave the optimum productivity of extracellular lipases was determined. Lyophilized extracellular enzymes were characterized by SDS-PAGE electrophoresis and isoelectric focusing (IEF). Depending on the nature of the carbon source, different isoenzymes were produced in various proportions. These samples were partially purified by different methodologies, including dialysis, adsorption chromatography and precipitation with ammonium sulfate or organic solvents. These characterizations allowed us to explain the relative catalytic activity of different samples, showing that in biocatalysis enzymes should not be treated simply as a »white magic powder« that can solve all the challenges in organic synthesis. Heptyl oleate synthesis, alcoxycarbonylation of amines and hydrolysis of the ester of ketoprofen are excellent reaction tests for the evaluation of lipase samples as biocatalysts.
Ključne riječi
lipase; isoenzymes; biocatalysis; purification of lipases; resolution of racemics
Hrčak ID:
110911
URI
Datum izdavanja:
15.12.2004.
Posjeta: 1.320 *