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Learning from Directed Evolution: Theoretical Investigations into Cooperative Mutations in Lipase Enantioselectivity
Author(s) -
Bocola Marco,
Otte Nikolaj,
Jaeger KarlErich,
Reetz Manfred T.,
Thiel Walter
Publication year - 2004
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200300731
Subject(s) - lipase , steric effects , molecular dynamics , hydrogen bond , enantiomer , chemistry , hydrolysis , mutant , directed evolution , stereochemistry , hydrophobic effect , hydrolase , enzyme , computational chemistry , molecule , biochemistry , organic chemistry , gene
Molecular modeling with classical force‐fields has been used to study the reactant complex and the tetrahedral intermediate in lipase‐catalyzed ester hydrolysis in 20 enzyme/substrate combinations. The R and S enantiomers of α‐methyldecanoic acid ester served as substrates for the wild‐type lipase from Pseudomonas aeruginosa and nine selected mutants. After suitable preparation of initial structures from an available wild‐type crystal structure, each system was subjected to 1 ns CHARMM force‐field molecular dynamics simulations. The resulting geometric and energetic changes allow interpretation of some experimentally observed effects of mutations, particularly with regard to the “hot spots” at residues 155 and 162. The replacement S155F enhances S enantiopreference through a steric relay involving Leu162. The double mutation S53P + L162G improves S enantioselectivity by creating a new binding pocket for the S enantiomer with an additional stabilizing hydrogen bond to His83. The simulations provide insight into remote and cooperative effects of mutations.