Influence of the hydrophobicity of lipase isoenzymes from Candida rugosa on its hydrolytic activity in reverse micelles.
Two isoenzymes of Candida rugosa lipase, having the same mol.wt., size and similar aminoacid sequence, were studied in reverse micelles of AOT. The results demonstrated the relevance of lipase hydrophobicity in reactions in anionic micelles. This is a key factor in mitigating the inhibition effect of charged micelles. The more hydrophobic isolipase A was a better biocatalyst for hydrolytic processes in these systems. Its alpha-helix content increased from 31% to 49% of the total structure in reverse micelles. A fluorescence study indicated a more apolar environment for the more hydrophobic isolipase A. Emission spectra of this isolipase in the AOT systems were blue shifted. At omega 0 values where each isolipase presented its maximum activity, a decrease of the emission intensity of Trp was found. An enzyme and substrate dependence of optimal omega 0 is reported. The different interaction of isolipases A and B with the micellar system produced an opposite omega 0 dependence to their stabilities. The more hydrophobic lipase A had higher stability at higher droplet sizes.
Otero
C
C
Instituto de Catálisis, CSIC, Cantoblanco, Madrid, Spain.
Rúa
M L
ML
Robledo
L
L
D002175:Candida / Q000201:enzymology*; D002942:Circular Dichroism; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D008823:Micelles; D012997:Solvents; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity
D007527:Isoenzymes; D008823:Micelles; D012997:Solvents; D008049:Lipase