6268
J . Org. Chem. 1997, 62, 6268-6273
A Th r ee-Dim en sion a l P r ed ictive Active Site Mod el for Lip a se fr om
P seu d om on a s cepa cia
Karin Lemke, Michael Lemke, and Fritz Theil*,†
Institut fu¨r Angewandte Chemie Berlin-Adlershof e.V., Rudower Chaussee 5, D-12484 Berlin, Germany
Received May 9, 1997X
A three-dimensional active site model of lipase from Pseudomonas cepaciasone of the most popular
lipases in organic synthesisswas developed on the basis of the kinetic resolution of 3-(aryloxy)pro-
pan-2-ols. Size and shape of both hydrophobic binding pockets of the active site of this lipase were
determined by substrate mapping in combination with molecular modeling for substrates and
nonsubstrates. This model explains and predicts whether a compound is accepted as a substrate
or not and allows to assess the enantiomer selectivity of the lipase-catalyzed reaction.
In tr od u ction
ester carbonyl groups. Furthermore, these structure
determinations show a common catalytic machinery for
all lipases. Despite this fact, substrate acceptance and
the degree of enantiodifferentiation is very different
depending upon the natural source of the lipase. It is
accepted in general that substrate recognition, stabiliza-
tion, and enantioselective transformation is determined
by two hydrophobic binding regions or pockets which are
not separated from the catalytic site.3f
One of the most popular lipases used in organic
synthesis is lipase from Pseudomonas cepacia from
Amano Pharmaceutical Co., Ltd. (Nagoya, J apan), called
lipase Amano PS. Before reidentification of the bacterial
source it was called lipase from P. fluorescens (lipase P).
Due to the flexibility of its binding sites lipase PS accepts
a broad range of substrates. This lipase has been used
for regio- and stereoselective hydrolysis4 and alcoholysis5
of carboxylic esters and anhydrides6 and for the regio-
and stereoselective transesterification of alcohols.4a,b,d,7
There seems to be almost no restriction regarding the
structure of compounds which are accepted as substrate
by lipase PS.
The use of enzymes has been established as an
important tool in organic synthesis during the past
decade. Due to their ability to discriminate between
enantiomers and enantiotopic groups, they are utilized
in kinetic resolutions of racemates and asymmetrizations
of prostereogenic or meso compounds to provide an easy
access to enantiomerically pure building blocks, synthetic
and natural products.1
Among the biocatalysts used in organic synthesis,
lipases (triacylglycerol hydrolases, EC 3.1.1.3) have been
used most frequently because they are cheap, available
from many sources, easy to handle, and accept a broad
range of substrates.2 Furthermore, they are active in
aqueous solution and in practically water-free organic
solvents. Particularly, in organic solvents lipases remain
their activity up to 100 °C. Lipases catalyze hydrolysis
and formation of carboxylic esters and formation of
amides upon the reaction conditions.
X-ray analyses of some lipases3 evidence that their
active sites are similar to those of serine proteases in
which the primary hydroxy function of serine of the
catalytic triade acts as a nucleophile to attack amide or
In order to rationalize and to predict reactivity and
selectivity of lipase-catalyzed biotransformations, a deeper
(4) (a) Klempier, N.; Faber, K.; Griengl, H. Synthesis 1989, 933. (b)
Foelsche, E.; Hickel, A.; Ho¨nig, H.; Seufer-Wasserthal, P. J . Org. Chem.
1990, 55, 1749. (c) Goergens, U.; Schneider, M. P. J . Chem. Soc., Chem.
Commun. 1991, 1064. (d) Goergens, U.; Schneider, M. P. J . Chem.
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Org. Chem. 1993, 58, 1054. (f) Hirose, Y.; Kariya, K.; Nakanishi, Y.;
Kurono, Y.; Achiwa, K. Tetrahedron Lett. 1995, 36, 1063. (g) Tanaka,
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Bianchi, D.; Bosetti, A.; Cesti, P.; Golini, P. Tetrahedron Lett. 1992,
33, 3231. (c) Koichi, Y.; Suginaka, K.; Yamamoto, Y. J . Chem. Soc.,
Perkin Trans. 1 1995, 1645.
† New address as of Sept. 1, 1997: University of Liverpool, Depart-
ment of Chemistry, P.O. Box 147, Liverpool L69 3BX, U.K.
X Abstract published in Advance ACS Abstracts, August 1, 1997.
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