Selectivity of celite-immobilized patatin (lipid acyl hydrolase) from potato (Solanum tuberosum L.) tubers in esterification reactions As influenced by water activity and glycerol analogues as alcohol acceptors.
Lipid acyl hydrolase (LAH; patatin) was purified from potato tubers by ammonium sulfate fractionation followed by anion-exchange and affinity chromatography. The major protein band of 40-43 kDa on SDS-PAGE appeared to be patatin, and it stained positive for lipase activity on native PAGE. Selectivity of a Celite-immobilized potato LAH in esterification reactions with n-acyl fatty acids (FA; C4, C6, C8, C10, C12, C14, C16, and C18) and alcohol acceptors (n-propanol, 2-propanol, 1,3-propanediol, and glycerol; 1,2-propanediol was not sufficiently reactive) was studied in isooctane. Immobilized LAH was highly selective for medium chain FAs (C8/C10) with a secondary optimum for chain lengths of C14/16. Water activity (a(w)) influenced activity and FA selectivity of the enzyme. Initial rates of ester synthesis were greatest at a(w) of 0.90 for all alcohol acceptors except for glycerol, where greatest initial rates were observed at a(w) of 0.19. Immobilized LAH preparations exhibited a bell-shape pH profile with optimum activity at pH 6-7 for ester synthesis, and no effect of pH on FA selectivity was observed.
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Department of Food Science, 1605 Linden Drive, University of Wisconsin, Madison, Wisconsin 53706, USA.
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D000438:Alcohols / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D007692:Diatomaceous Earth; D004591:Electrophoresis, Polyacrylamide Gel; D005227:Fatty Acids / Q000378:metabolism; D005990:Glycerol / Q000031:analogs & derivatives*; D010940:Plant Proteins / Q000378:metabolism*; D011198:Solanum tuberosum / Q000201:enzymology*; D014867:Water
D000438:Alcohols; D005227:Fatty Acids; D010940:Plant Proteins; C043222:patatin protein, Solanum tuberosum; D014867:Water; D007692:Diatomaceous Earth; D002265:Carboxylic Ester Hydrolases; D005990:Glycerol