Effects of organic solvents on immobilized lipase in pectin microspheres.
Lipase from Brevibacillus agri 52 was found stable up to 90% diethylenglycol (DEG), glycerol (GLY), and 1,2 propanediol (1,2 PRO) at 37 degrees C for 1 h and the stability was reduced only approximately 20% after 12 h incubation, but in 40% dimethylsulfoxide (DMSO), lipase activity was stable only for 1 h. Inhibition of the biocatalysts with dimethylformamide (DMF) was detected at 20% solvent concentration. In water immiscible systems, the stability of lipase in n-hexane, n-tetradecane and n-heptane resembles the water activity, but in the presence of isobutanol, 1-hexanol, and butylbutirate, the stability was significantly reduced. Lipase 52 precipitates in the presence of 50% acetone or ethanol/water mixtures, but enzymatic activity was partially recovered by adding 20% GLY, DEG, 1,2 PRO, or DMSO to the reaction mixture. Furthermore, by increasing DEG in 70% DMF/DEG mixtures, the lipase activity was protected. Encapsulation of lipase in pectin gels cross-linked with calcium ions brings three to four times more enzymatic activity in 70% water miscible organic solvents compared to aqueous systems.
Facultad de Ciencias Exactas, CINDEFI, Institute of Applied Biotechnology and INIFTA, UNLP-CCT La Plata-CONICET, Universidad Nacional de La Plata, La Plata, Argentina.
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D000438:Alcohols / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004126:Dimethylformamide / Q000494:pharmacology; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000187:drug effects*; D005026:Ethylene Glycols / Q000494:pharmacology; D005990:Glycerol / Q000494:pharmacology; D016937:Gram-Positive Endospore-Forming Rods / Q000201:enzymology; D008049:Lipase / Q000037:antagonists & inhibitors / Q000187:drug effects*; D008863:Microspheres; D010368:Pectins; D019946:Propylene Glycol / Q000494:pharmacology; D012997:Solvents / Q000494:pharmacology*
D000438:Alcohols; D004800:Enzymes, Immobilized; D005026:Ethylene Glycols; D012997:Solvents; C013484:diethylene glycol; D019946:Propylene Glycol; D004126:Dimethylformamide; D010368:Pectins; D008049:Lipase; D005990:Glycerol; D004121:Dimethyl Sulfoxide