Condensed Tannins: A Proposed Route to 2R,3R-(2,3-cis)-Proanthocyanidins

Richard W. Hemingway. and Peter E. Laksb ..

a Southern Forest Experiment Station, USDA-Forest Service, 2500 Shreveport Highway, Pineville, Louisiana 71360,
U.S.A.
b Institute of Wood Research, Michigan Technological University, Houghton, Michigan 49931, U.S.A.

The biogenesisof 2R,3R-(2,3-cis)-proanthocyanidins

from 2R,3R-(2,3-trans)-dihydrofiavonols
can be accountedfo!
by tautomerism between quinone methide and flav-3-en-3-olintermediates.

Reprinted from the Journal of The Chemical Society

Chemical Communications 1985
746 J. CHEM.sac CHEM.COMMUN 1985

Condensed Tannins: A Proposed Route to 2R,3R-(2,3-c;s)-Proanthocyanidins

Richard W. Hemingway. and Peter E. Laksb
a Southern ForestExperiment Station, USDA-ForestService,2500 Shreveport Highway, Pineville, Louisiana 71360,
U.S.A.
b Institute of Wood Research,Michigan Technological University, Houghton, Michigan 49931, U.S.A.

The biogenesisof 2R,3R-(2,3-cis)-proanthocyanidins

from 2R,3R-(2,3-trans)-dihydrofiavonols
can be accounted for
by tautomerism between quinone methide and flav-3-en-3-olintermediates.

ROUX,IHaslam,2-4 and Stafford5--8have proposed differing

biogenetic sc~emesfor the formation of proanthocyanidins
(condensedtannins) in plants. All three schemessuffer from
difficulties in explaining the formation of the 2R,3R-(2,3-cis)-
proanthocyanidins that predominate in plant tannins. Staf-
ford5--8 has postulated that a C-3 epimerase acts on (+)-
dihydroquercetin to provide the 2R ,3S-(2,3-cis)-di-
hydroflavanol that would then be reduced to the correspond-
ing 2R,3R,4S- (or 4R-) flavan-3.4-diol. However, enzyme
preparations from Douglas-fir callus do not produce any

,/
2,3-cis-procyanidins.5--8Haslam2-4 proposed a flav-3-en-3-o1
--
~
(1)
intermediate that might be derived from an a-hydroxychal-
cone. A stereospecificreduction of the flav-3-en-3-01to either OH OH
2,3-cis-or 2,3-trans-flavan-4-carbocationsisthen postulated to O~H ~OOH OH 4'
account for the formation of the proanthocyanidins. but there
is debate about the presenceof a carbocation. ROUXIproposes \~.A,."'~I""J~
S. S .
OH
5
that the a-hydroxychalcone can give either 2.3-cis- or 2,3-
trans-dihydroflavonols through the familiar chalcone- CHz CHZ CHz
Ph. Ph Ph
flavonone interconversion. but the stereochemistry of the
dihydroflavonols in many plants is 2,3-trans. while that of the
co-occurring proanthocyanidins is predominantly 2 .3-cis,asin
Douglas-fir .5-9
Procyanidins are readily synthesised from flavan-3,4-diols
by mild base-or acid-catalysedgeneration of quinone methide
0
I ~ ~ ~
OH
~t;JJ°H~ I OHHO
~H OH ~
OH

~~
H
I OH
HO>~~ H
OH

h
cO

intermediates.I~12 Platt et al.4 have alluded to the tautomeric

relationships of these quinone methides with flav-3-en-3-0Is. 5
Chemical evidence supporting these interconversions was !r CHz
Ph tt
obtained in recent studies of the reactions of condensed
tannins in alkaline solutions.13 At pH 12.0 and ambient
temperature. in the presenceof an excessof phenylmethane-
thiol, procyanidins (I) are cleaved to form mono- or di-
benzylsulphide derivatives from the lower and upper units,
Q~~r'(JII
HO
~
OH OH
~ ~
OH
H HO~OH
~)1
<?H
~",,~
~OH
OH

respectively. Loss of phenylmethanethiol and tautomeric OH OH

rearrangement leads to the formation of two propan-2-one
derivatives, (2} and (3) (Scheme 1). These ketones are CHz
Ii Ph l1
analogous to Haslam's2-4 proposed flav-3-en-3-ol. which, in
solution, would be in the keto form. Since these reactions are
at high pH, all products are produced through cleavageof the OH OH
pyran ring. However, it seems plausible that enzyme- lQf ~Jt"rM'
H 0 '/ I HOr?'J1OH R r"'"']I°H
catalysed reactions on flavan-3,4-diols could involve the
formation of a quinone methide (6)1(1-12followed by a ~ ~ ~,~A)J
tautomeric rearrangement analogous to the experimentally
OH OH
observed reactions described in Scheme 1 to produce the 5
CHz (3)
flav-3-en-3-01or its corresponding ketone without cleavageof Ph
the heterocyclic ring. (2)
Based on these results, the following explanation is pro-
posed for the biogenesis of either 2.3-cis- or 2.3-trans- Scheme1. Ketones produced from the reaction of procyanidins with
proanthocyanidins from 2R.3R-(2,3-trans)-dihydroflavanols phenylmethanetbiol and base at ambient temperature.
(Scheme2). In agreement with Stafford.5-11enzymatic reduc-
tion of the dihydroflavonol (4) provides the 2R.3S,4S-flavan-
3,4-diol (5). But rather than the formation of a flavan-4-
carbocation, the diol eliminates water to form a 2,3-trans- catalysed tautomerism observed in the reactions of the
quinone methide (6). In agreement with Haslam.I-3 it is diarylpropanoidsl3 (Scheme 1). The flav-3-en-3-01 is then
proposed that a flav-3-en-3-ol is a central intermediate. This stereospecifically converted back into a quinone methide (6)
compound (7) is formed from the quinone metbide by an or to (8) that will then polymerise to form the condensed
enzyme-catalysed rearrangement parallel to the base- tannins (9). Rearrangements()f the flavan-3-0Is analogousto
J. CHEM. SOC., CHEM. COMMUN 1985 747

OH

Ql O ,(:DOH

HO
~I ,..H

OH OH

(4)

~°"r'&
~,~
OH x~OH
(6)

¥
t~

~ OH
~H
'"'r""'¥ 0'r ,-0""
~...~A HO«I'l.-JJ
OHOH ~~...~~
(1)
(9)
'\
11
""
OM

ow o ,(y'
~ ~ ..OH
H
(8)

Scheme2. Proposed route for biosynthesis of proanthocyanidins.

the formation of the ketone (3) (Scheme 1) could account for References
the formation of 2,3-cis-fiavan-3-olsfrom a two step reduction 1 D. G. Roux and D. Ferreira. Phytochemistry. 1974.13;2039.
2 D. Jacques,C. T. Opie. L. J. Porter, and E. Haslam,J. Chern.
of the 2,3-trans-dihydroflavanols.
This proposal is in full agreement with Haslam's labelling Soc.,Perkin Trans.I, 1977.1637.
studies that show retention of the proton at C-2 and loss of a 3 E. Haslam,Phytochemis,ry, 1977,16. 1{'25.
4 R. V. Platt,C. T. Opie.andE. Haslam.Ph.vtochemistry,1984,23,
proton at C-3 of the fIavan unit.2-4 By assuming different
enzyme sites for the synthesis of the procyanidin and 2211.
5 H. A. StaffordandH. H. Lester, Plan' Physiol..1981,68.1035.
fIavan-3-01units, one can account for the differences in their 6 H. A. Stafford.Phytochemistry,1983.22.2643.
hydroxylation and/or stereochemistry. Most importantly, 7 H. A. Staffordand H. H. Lester.PlantPhysiol..1984.76.184.
interconversions of the quinone methides and compounds 9 J.1. Karchesy. P. M. Loveland. M. L. Laver, D. F. Barofsky. and
8
H.
A.
Stafford
and
L.
J.
Porter.
Phytochemistry,
in
the
pres
analogous to fIav-3-en-3-0Is have been demonstrated chem-
ically in the formation of (2) and (3) from reactions of E. Barofsky, Phytochemistry, 1976. IS. 200).
10 R. W. Hemingway and l. Y. Foo, J. Chern. Soc., Chern.
condensed tannins with phenylmethanethiol at alkaline pH.
Our results suggest that one should look for enzymes Commun., 1983. l-O35.
11 L. Y. Foo and R. W. Hemingway. J. Chern. Soc., Chern.
mediating these interconversions rather than for dihydro-
Commun.. 1984,85.
flavonol C-3 epimerases. 12 M. R. Attwood, B. R. Brown, S. G. Lisseter. C. L. Torrero. and
P. M. Weaver. J. Chern. Soc., Chern. Commun., 1984.177.
13 P. E. Laks and~. W. Hemingway. manuscript in preparation.
Reaived. 11th February 1'985;Com. 170