Influence of delta-functional groups on the enantiorecognition of secondary alcohols by Candida antarctica lipase B.
The selectivity of acetylation of delta-functionalized secondary alcohols catalyzed by Candida antarctica lipase B has been examined by molecular dynamics. The results from the simulation show that a delta-alcohol functionality forms a hydrogen bond with the carbonyl group of Thr 40. This interaction stabilizes the tetrahedral intermediate and thus leads to selective acetylation of the R enantiomer. A stabilizing interaction of the delta-(R)-acetoxy group with the peptide NH of alanine 282 was also observed. No stabilizing interaction could be found for the delta-keto functionality, and it is proposed that this is the reason for the experimentally observed decrease in enantioselectivity. From these results, it was hypothesized that the enantioselectivity could be restored by mutating the alanine in position 281 for serine. The mutation was made experimentally, and the results show that the E value increased from 9 to 120.
Nyhlén
Jonas
J
Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, 106 91 Stockholm, Sweden.
Martín-Matute
Belén
B
Sandström
Anders G
AG
Bocola
Marco
M
Bäckvall
Jan-E
JE
D000085:Acetates / Q000378:metabolism; D000438:Alcohols / Q000737:chemistry* / Q000378:metabolism*; D002384:Catalysis; D004951:Esterification; D005656:Fungal Proteins; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D011487:Protein Conformation; D013237:Stereoisomerism; D013379:Substrate Specificity
D000085:Acetates; D000438:Alcohols; D005656:Fungal Proteins; D008049:Lipase; C431349:lipase B, Candida antarctica