Kinetics of p-nitrophenyl acetate hydrolysis catalyzed by Mucor javanicus lipase in AOT reverse micellar solutions formulated in different organic solvents.
The rate of hydrolysis of p-nitrophenyl acetate (PNPA) catalyzed by Mucor javanicus lipase has been measured in AOT reverse micellar solutions formulated in aliphatic hydrocarbons, aromatic hydrocarbons and a chlorinated compound. The study has been performed at a single value of W=([water]/[AOT])=6.0. Fluorescence decay measurements of intrinsic enzyme fluorescence, mainly due to tryptophan residues, in the different reverse micellar systems were also carried out, in an attempt to obtain some insight on the effect of the organic solvent on the protein conformation. Differences observed in the kinetics of the fluorescence decays of tryptophan residues of the lipase incorporated to the micelles with the different external organic solvents were also found in the catalytic behaviour of the enzyme. In particular, it is observed that the contribution of the long lived component of the fluorescence decay is considerably higher (ca. 40%) in aliphatic than in aromatic solvents (ca. 15%), indicating significant differences in the protein conformation. This effect of the organic solvent on the protein conformation is also observed in the enzymatic activity, which is higher in the aromatic than in the aliphatic solvents.
Abuin
Elsa
E
Facultad de Química y Biología, Universidad de Santiago de Chile, Casilla 40-Correo 33, Santiago, Chile. eabuin@lauca.usach.cl
Lissi
Eduardo
E
Biasutti
M Alicia
MA
Duarte
Roxanna
R
D002384:Catalysis; D006868:Hydrolysis; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D008823:Micelles; D009089:Mucor / Q000201:enzymology*; D009596:Nitrophenols / Q000737:chemistry*; D009930:Organic Chemicals / Q000737:chemistry / Q000378:metabolism; D012997:Solvents / Q000737:chemistry*; D013386:Succinates / Q000737:chemistry* / Q000378:metabolism
C512554:1,4-bis(2-ethylhexyl) sodium sulfosuccinate; D008823:Micelles; D009596:Nitrophenols; D009930:Organic Chemicals; D012997:Solvents; D013386:Succinates; C008642:4-nitrophenyl acetate; D008049:Lipase