Alcohol inhibition and specificity studies of lipase B from candida antarctica in organic solvents

Alcohol inhibition of the lipase B from Candida antarctica has been studied through two different approaches: using the same inhibitor (1-butanol) in different organic solvents and using different inhibitors (differing in chain length) in the same solvent. The competitive inhibition constant values obtained in each case correlate with the calculated activity coefficients of the substrate, suggesting that desolvation of the alcohol is the major force changed. Data dispersion observed using the second approach has been interpreted to come from contributions of enzyme-inhibitor interactions to the binding energy. On the other hand, deacylation has been found to be much less influenced by the solvent variation than the acylation step, despite of the fact that solvation of the substrate involved in this step (the alcohol) is expected to change more than for the ester. Concerning the specificity behavior of the enzyme, a bimodal pattern was observed for the deacylation rate dependence on the alcohol chain length, with the highest values for hexanol (C6) and decanol (C10). With regard to the ester specificity, ethyl caproate (C6) is the preferred one. These results have been confronted with those reported for the lipase from Candida rugosa. Copyright 1998 John Wiley & Sons, Inc.

[1]  P. Wangikar,et al.  Active‐site titration of serine proteases in organic solvents , 1996, Biotechnology and bioengineering.

[2]  A. Klibanov,et al.  THE MECHANISTIC DISSECTION OF THE PLUNGE IN ENZYMATIC ACTIVITY UPON TRANSITION FROM WATER TO ANHYDROUS SOLVENTS , 1996 .

[3]  P. Halling,et al.  Substrate specificity and kinetics of Candida rugosa lipase in organic media. , 1996, Enzyme and microbial technology.

[4]  G J Kleywegt,et al.  Crystallographic and molecular-modeling studies of lipase B from Candida antarctica reveal a stereospecificity pocket for secondary alcohols. , 1995, Biochemistry.

[5]  P. Wangikar,et al.  Probing enzymic transition state hydrophobicities. , 1995, Biochemistry.

[6]  J. Jongejan,et al.  Do organic solvents affect the catalytic properties of lipase? Intrinsic kinetic parameters of lipases in ester hydrolysis and formation in various organic solvents , 1995, Biotechnology and bioengineering.

[7]  Miroslaw Cygler,et al.  A Structural Basis for the Chiral Preferences of Lipases , 1995 .

[8]  P. Halling,et al.  Specificities of Enzymes Corrected for Solvation Depend on the Choice of the Standard State , 1994 .

[9]  R. Kazlauskas,et al.  Elucidating structure-mechanism relationships in lipases: prospects for predicting and engineering catalytic properties. , 1994, Trends in biotechnology.

[10]  D. Clark,et al.  Transition state stabilization of subtilisins in organic media , 1994, Biotechnology and bioengineering.

[11]  P. Wangikar,et al.  Protein and solvent engineering of subtilisin BPN' in nearly anhydrous organic media , 1993 .

[12]  A. Russell,et al.  Kinetic analysis of the mechanism for subtilisin in essentially anhydrous organic solvents , 1993 .

[13]  V. Partali,et al.  Enzymatic resolution of butanoic esters of 1-phenyl, 1-phenylmethyl, 1-[2-phenylethyl] and 1-12-phenoxyethyl] ethers of 3-methoxy-1,2-propanediol , 1993 .

[14]  J. Dordick,et al.  Tailoring lipase specificity by solvent and substrate chemistries , 1993 .

[15]  A. Klibanov,et al.  Solvent variation inverts substrate specificity of an enzyme , 1993 .

[16]  K. Hult,et al.  S-ethyl thiooctanoate as acyl donor in lipase catalysed resolution of secondary alcohols , 1993 .

[17]  J. Baratti,et al.  Multi-competitive enzymatic reactions in organic media: Application to the determination of lipase alcohol specificity , 1992 .

[18]  V. Gotor,et al.  Vinyl carbonates as novel alkoxycarbonylation reagents in enzymatic synthesis of carbonates. , 1992 .

[19]  A. Russell,et al.  Determination of equilibrium and individual rate constants for subtilisin‐catalyzed transesterification in anhydrous environments , 1992, Biotechnology and bioengineering.

[20]  K. Breddam,et al.  Interdependency of the binding subsites in subtilisin. , 1992, Biochemistry.

[21]  M. Reuss,et al.  A kinetic study of immobilized lipase catalysing the synthesis of isoamyl acetate by transesterification in n-hexane. , 1992, Enzyme and microbial technology.

[22]  F. Malcata,et al.  Kinetics and mechanisms of reactions catalysed by immobilized lipases. , 1992, Enzyme and microbial technology.

[23]  J. Dordick,et al.  How do organic solvents affect peroxidase structure and function? , 1992, Biochemistry.

[24]  Aviva Rappaport,et al.  A rule to predict which enantiomer of a secondary alcohol reacts faster in reactions catalyzed by cholesterol esterase, lipase from Pseudomonas cepacia, and lipase from Candida rugosa , 1991 .

[25]  A. Klibanov,et al.  Kinetic isotope effect investigation of enzyme mechanisms in organic solvents , 1990 .

[26]  A. Klibanov,et al.  Hammett analysis of enzyme action in organic solvents , 1989 .

[27]  J. Baratti,et al.  Multi-competitive enzymatic reactions in organic media: a simple test for the determination of lipase fatty acid specificity. , 1989, Biochimica et biophysica acta.

[28]  J. Kraut,et al.  How do enzymes work? , 1988, Science.

[29]  T. Sakurai,et al.  Control of enzyme enantioselectivity by the reaction medium , 1988 .

[30]  A. Klibanov,et al.  Enzymatic catalysis in nonaqueous solvents. , 1988, The Journal of biological chemistry.

[31]  Alexander M. Klibanov,et al.  Enzyme-catalyzed processes in organic solvents. , 1985 .

[32]  K. Kawai,et al.  Microbially mediated enantioselective ester hydrolyses utilizing Rhizopus nigricans. A new method of assigning the absolute stereochemistry of acyclic 1-arylalkanols , 1983 .