J . Org. Chem. 1997, 62, 7841-7843
7841
En zym a tic Resolu tion of Ch ir a l 2-Hyd r oxy Ca r boxylic Acid s by
En a n tioselective Oxid a tion w ith Molecu la r Oxygen Ca ta lyzed by
th e Glycola te Oxid a se fr om Sp in a ch (Spin a cia oler a cea )
Waldemar Adam,*,† Michael Lazarus,‡ Barbara Boss,‡ Chantu R. Saha-Mo¨ller,†
Hans-Ulrich Humpf,‡ and Peter Schreier‡
Institute of Organic Chemistry and the Institute of Pharmacy and Food Chemistry,
University of Wu¨rzburg, Am Hubland, D-97074 Wu¨rzburg, Germany
Received J uly 15, 1997X
The enzymatic oxidation of a variety of saturated and unsaturated aliphatic derivatives of racemic
2-hydroxy acids 1 to their 2-oxo acids 2 with molecular oxygen catalyzed by the glycolate oxidase
from spinach (Spinacia oleracea) was shown to proceed highly enantioselectively. Thus, the glycolate
oxidase-catalyzed kinetic resolution provides a convenient biocatalytic method for the preparation
of enantiomerically pure (R)-2-hydroxy acids. The absolute configuration of the (R)-2-hydroxy acid
1b was assigned by comparison of the measured optical rotation value with that of the literature
data and by application of the exciton-coupled circular dichroism method (ECCD) on its bichro-
mophoric 2-naphthoate 9-methylanthryl derivative 3b. These results establish the ECCD method
as a convenient microscale chirooptic tool for the configurational assignment of 2-hydroxy acids.
Sch em e 1. Oxid a tion of 2-Hyd r oxy Acid s w ith
Molecu la r Oxygen Ca ta lyzed by Glycola te Oxid a se
fr om Sp in a ch (S. oler a cea )
In tr od u ction
The glycolate oxidase (EC 1.1.3.15) is a peroxisomal
enzyme, which is found in the leaves of many green
plants and in the liver of mammalians. This enzyme has
been isolated from a variety of biological sources, includ-
ing the leaves of spinach,1a pea,1b pumpkin,1c and cucum-
ber cotyledons1d and the liver of pigs,1e rats,1f and
humans.1g,h The glycolate oxidase was reported to be
catalytically active only as tetramers or octamers of
identical subunits, which have a molecular mass of
43 000. The extensively studied enzyme has been al-
ready expressed in genetically manipulated micro-
organisms.2a,b The enzyme converts glycolic acid to
glyoxylic acid (Scheme 1, R ) H) and has been employed
in organic synthesis as a biocatalyst for the production
of glyoxylic acid by using the soluble enzyme,3a the
enzyme immobilized on a solid support,3b or the geneti-
cally engineered protein.3c Already in the 1960s, it was
reported4a,b that the glycolate oxidase accepts also 2-hy-
droxy carboxylic acids as substrates, and the kinetic
studies revealed that L-lactic acid is preferentially con-
verted by the catalytically active protein.4a However, to
date there is no report on the preparation of optically
active 2-hydroxy acids by enantioselective oxidation of
racemic acids with molecular oxygen catalyzed by the
glycolate oxidase.
* Corresponding author. Fax: 0049-931-8884756. E-mail: Adam@
uni-wuerzburg.de.
Optically active 2-hydroxy acids are important building
blocks for the asymmetric synthesis of glycols,5a halo
esters,5b and epoxides.5c Several chemical6 and enzy-
matic7,8 methods have been reported previously on the
synthesis of optically active R-hydroxy acids. The enzy-
† Institute of Organic Chemistry.
‡ Institute of Pharmacy and Food Chemistry.
X Abstract published in Advance ACS Abstracts, October 1, 1997.
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