Optimization of Pseudomonas cepacia lipase preparations for catalysis in organic solvents.
The activity of different lipase (from Pseudomonas cepacia) forms, such as crude powder (crude PC), purified and lyophilized with PEG (PEG + PC), covalently linked to PEG (PEG-PC), cross-linked enzyme crystals (CLEC-PC), and immobilized in Sol-Gel-AK (Sol-Gel-AK-PC) was determined, at various water activities (aw), in carbon tetrachloride, benzene and 1,4-dioxane. The reaction of vinyl butyrate with 1-octanol was employed as a model and both transesterification (formation of 1-octyl butyrate) and hydrolysis (formation of butyric acid from vinyl butyrate) rates were determined. Both rates depended on the lipase form, solvent employed, and aw value. Hydrolysis rates always increased as a function of aw, while the optimum of aw for transesterification depended on the enzyme form and nature of the solvent. At proper aw, some lipase forms such as PEG + PC, PEG-PC, and Sol-Gel-AK-PC had a total activity in organic solvents (transesterification plus hydrolysis) which was close to (39 and 48%) or even higher than (130%) that displayed by the same amount of lipase protein in the hydrolysis of tributyrin-one of the substrates most commonly used as standard for the assay of lipase activity-in aqueous buffer. Instead, CLEC-PC and crude PC were much less active in organic solvents (2 and 12%) than in buffer. The results suggest that enzyme dispersion and/or proper enzyme conformation (favored by interaction with PEG or the hydrophobic Sol-Gel-AK matrix) are essential for the expression of high lipase activity in organic media.
Secundo
F
F
Istituto di Biocatalisi e Riconoscimento Molecolare, via Mario Bianco 9, Milano 20131, Italy. secundo@ico.mi.cnr.it
Spadaro
S
S
Carrea
G
G
Overbeeke
P L
PL
D001709:Biotechnology; D002021:Buffers; D016956:Burkholderia cepacia / Q000201:enzymology*; D002384:Catalysis; D004951:Esterification; D006868:Hydrolysis; D008049:Lipase / Q000302:isolation & purification* / Q000378:metabolism; D011092:Polyethylene Glycols; D012997:Solvents; D014867:Water
D002021:Buffers; D012997:Solvents; D014867:Water; D011092:Polyethylene Glycols; D008049:Lipase