Improved catalytic properties of immobilized lipases by the presence of very low concentrations of detergents in the reaction medium.
The addition of a very small concentration of a detergent (in many instances under the critical micellar concentration (cmc)) has been found to greatly increase the activity of immobilized lipases, using those from Pseudomonas fluorescens (PFL) and Candida antarctica (isoform B) as model enzymes. However, the detergents may also have a negative effect on enzyme activity; in fact, for all enzyme preparations and substrates the activity/detergent concentration curve reached a maximum value and started to decrease, in many instances even under the initial value. The concentration and nature of the detergent (SDS, CTAB, Triton X-100, or X-45) that permitted the maximum hyperactivation was different depending on the substrate. The best hyperactivation values promoted by the presence of detergent were over a 20-fold factor. The presence of detergents permitted the inhibition of lipases by irreversible covalent inhibitors (e.g., 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride) (AEBSF) while the enzyme, in the absence of detergent, is not inhibited by these irreversible inhibitors. This suggested that the main effect of the detergents is to shift the conformational equilibrium of lipases toward the open form. Moreover, the presence of detergents also permitted to improve the enantioselectivity exhibited by the immobilized lipases in some cases. For example, the enantioselectivity of PFL-glyoxyl agarose increased from 40 to more than 100 in the hydrolysis of (+/-)-2-hydroxy-4-phenylbutyric acid ethyl ester by using 0.1% CTAB.
Fernandez-Lorente
Gloria
G
Departamento de Biocatálisis, Instituto de Catálisis (CSIC), Campus UAM Cantoblanco, 28049 Madrid, Spain.
Palomo
Jose M
JM
Cabrera
Zaida
Z
Fernandez-Lafuente
Roberto
R
Guisán
José M
JM
D001672:Biocompatible Materials / Q000737:chemistry*; D001709:Biotechnology / Q000379:methods*; D002384:Catalysis; D003902:Detergents / Q000032:analysis / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000737:chemistry*; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D011551:Pseudomonas fluorescens / Q000201:enzymology
D001672:Biocompatible Materials; D003902:Detergents; D004800:Enzymes, Immobilized; D008049:Lipase