Mechanism of enantioselectivity of lipases and other synthetically useful hydrolases

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Abstract

Lipases are biocatalysts most widely used in organic synthesis. Unusual but attractive feature of lipases is that, in addition to high catalytic activity and thermostability in organic solvents, lipases show high enantioselectivity and broad substrate specificity simultaneously. They show excellent enantioselectivity especially toward a wide range of secondary alcohols. The mechanistic details or stereoselective organic reactions are relatively well understood, and knowledge has been used to develop new chemical reagents. In contrast, biocatalysts are behind chemical reagents in rational design approaches, which is partly due to the mechanistic ambiguity or enzymatic reactions. The mechanistic aspects of enantioselective biocatalysts are nevertheless becoming clear. This review provides an overview of the studies aimed at understanding the mechanisms of enantioselectivity of synthetically useful hydrolases such as lipases, subtilisins and chymotrypsins toward unnatural chiral substrates. Several methods for addressing the mechanism are introduced: (i) substrate mapping, (ii) X-ray crystallographic analysis, (iii) computational calculations, (iv) kinetic analysis, (v) thermodynamic analysis, (vi) site-directed or random mutagenesis, (vii) spectroscopic methods such as fluorescence, ESR, and mass spectroscopy. Different models and mechanisms proposed so far are selected and explained. The chemical principles revealed by the mechanistic studies will be useful for (i) using the enzymes in organic synthesis efficiently, (ii) altering the features or the enzymes rationally, (iii) utilizing them as a tool for determining the absolute stereochemistry of molecules, and (iv) designing new artificial catalysts mimicking the catalytic machinery of the enzymes.

Original languageEnglish
Pages (from-to)1009-1025
Number of pages17
JournalCurrent Organic Chemistry
Volume8
Issue number11
DOIs
Publication statusPublished - Jul 2004

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Enantioselectivity
Hydrolases
Lipase
Enzymes
Substrates
Subtilisins
Mutagenesis
Stereochemistry
Chymotrypsin
Organic solvents
Machinery
Paramagnetic resonance
Catalyst activity
Fluorescence
Alcohols
Spectroscopy
Thermodynamics
X rays
Catalysts
Molecules

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Mechanism of enantioselectivity of lipases and other synthetically useful hydrolases. / Ema, Tadashi.

In: Current Organic Chemistry, Vol. 8, No. 11, 07.2004, p. 1009-1025.

Research output: Contribution to journalArticle

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