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Appl Environ Microbiol, June 1998, p. 2026-2031, Vol. 64, No. 6
Microbiology Department, University College,
Cork, Ireland,1 and
Division of
Industrial Microbiology, Department of Food Science, Wageningen
Agricultural University, 6700 EV Wageningen, The
Netherlands2
Received 29 January 1998/Accepted 23 March 1998
Laccase is a copper-containing phenoloxidase, involved in lignin
degradation by white rot fungi. The laccase substrate range can be
extended to include nonphenolic lignin subunits in the presence of a
noncatalytic cooxidant such as
2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) (ABTS), with ABTS
being oxidized to the stable cation radical, ABTS·+,
which accumulates. In this report, we demonstrate that the
ABTS·+ can be efficiently reduced back to ABTS
by physiologically occurring organic acids such as oxalate, glyoxylate,
and malonate. The reduction of the radical by oxalate results in the
formation of H2O2, indicating the formation of
O2·
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Reduction of the 2,2'-Azinobis(3-Ethylbenzthiazoline-6-Sulfonate)
Cation Radical by Physiological Organic Acids in the Absence and
Presence of Manganese
as an intermediate.
O2·
itself was shown to act as an
ABTS·+ reductant. ABTS·+ reduction and
H2O2 formation are strongly stimulated by the
presence of Mn2+, with accumulation of Mn3+
being observed. Additionally,
4-methyl-O-isoeugenol, an unsaturated lignin monomer model,
is capable of directly reducing ABTS·+. These data
suggest several mechanisms for the reduction of ABTS·+
which would permit the effective use of ABTS as a laccase cooxidant at
catalytic concentrations.
*
Corresponding author. Mailing address: Microbiology
Department, University College, Cork, Ireland. Phone: 353-21-902743. Fax: 353-21-903101. E-mail: a.dobson{at}ucc.ie.
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