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Rational design of enantioselective enzymes requires considerations of entropy
Author(s) -
Ottosson Jenny,
RotticciMulder Johanna C.,
Rotticci Didier,
Hult Karl
Publication year - 2001
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.13501
Subject(s) - delta , enantioselective synthesis , chemistry , enthalpy , enantiomer , configuration entropy , entropy (arrow of time) , candida antarctica , enzyme , stereochemistry , thermodynamics , physics , lipase , biochemistry , catalysis , astronomy
Abstract Entropy was shown to play an equally important role as enthalpy for how enantioselectivity changes when redesigning an enzyme. By studying the temperature dependence of the enantiomeric ratio E of an enantioselective enzyme, its differential activation enthalpy (Δ R‐S Δ H ‡ ) and entropy (Δ R‐S Δ S ‡ ) components can be determined. This was done for the resolution of 3‐methyl‐2‐butanol catalyzed by Candida antarctica lipase B and five variants with one or two point mutations. Δ R‐S Δ S ‡ was in all cases equally significant as Δ R‐S Δ H ‡ to E. One variant, T103G, displayed an increase in E, the others a decrease. The altered enantioselectivities of the variants were all related to simultaneous changes in Δ R‐S Δ H ‡ and Δ R‐S Δ S ‡ . Although the changes in Δ R‐S Δ H ‡ and Δ R‐S Δ S ‡ were of a compensatory nature the compensation was not perfect, thereby allowing modifications of E. Both the W104H and the T103G variants displayed larger Δ R‐S Δ H ‡ than wild type but exhibited a decrease or increase, respectively, in E due to their different relative increase in Δ R‐S Δ S ‡ .