Decarboxylation of a-Keto Acids
R. W. Hanson
Department of Biological Sciences, Plymouth Polytechnic, Plymouth. Devon, UK
It is well established that some of the most important
reactions that a-keto acids, RCOCOzH (I), undergo during
metabolism involve decarhoxylation ( I ) . For example pyru-
vic acid (1, R = CH3) is decarhoxylated nonoxidatively dur-
ing alcoholic fermentation
PD
CH,COCO, + Hi * CH,CHO + CO,
(PD = pyruvate decarhoxylase) and in the biosyntbesis of
acyloins such as acetolactate,
ALS
2CH,COC02 + Ht = CH,COC(CH),(OHICO,- + CO,
(A1.S = acetolacrate synrhetase~.Additionally, pvruvir arid
i c r l . R = -O,CCI-IqCH?)
nn(l n - k p t w l ~ ~ t n racid . . undwzo oxi-
dative dec~rhoxylationwhen they-are converted to acetyl-
coenzyme A and succinylcoenzyme A, respectively,
CH,COCO,- + NADt + HSCoA
PDH
+ CH,COSCoA + NADH + CO,
~O,CCH,CH,COCO,- + NADt + HSCoA
-
a-KGDH
-O,CCH,CH,COSCoA + NADH + CO,
(PDH = pyruvate dehydrogenase complex; a-KGDH = a -
ketoglutarate dehydrogenase complex). The enzymes that
catalyze reactions 1-4 all have an absolute requirement for
thiamine pyrophosphate (TPP),
as coenzyme. Almost every undergraduate textbook of hio-
chemistry contains an explanation of how T P P is involved in
the decarboxylation of a-keto acids, hut the explanation is
rarely accompanied by a comparative discussion of analo-
gous nonenzymic reactions, which is essential if the student
is to gain a proper understanding of the chemical role of the
coenzyme.
Unfortunately, information regarding the nonenzymic de-
carhoxylation of ru-keto acids is not readily available. The
recent review of the synthesis and properties of a-keto acids
by Cooper, Ginos, and Meister ( 2 ) might he expected to
provide such information. Three of the four papers cited in
the review under the heading "thermal decarboxylation"
refer, however, to decarhoxylations catalyzed by metal ions
(the fourth refers to catalysis by cyanide, below). The mech-
anisms of these reactions are unclear and therefore provide
no insight into the mode of action of TPP. The situation
regarding textbooks is even worse. Most textbooks of organic
chemistry ignore a-keto acids entirely or make no specific
reference to their decarhoxylation. The logical conclusion to
be drawn from this is that the nonenzymic decarboxylation
of a-keto acids is difficult to accomplish and has not been
investigated extensively.' At least two authors, however,
make statements which contradict this. Thus March (3)
includes wketo acids in a table of "some acids which under-
go decarhoxylation fairly easily", while Finar (4) is more
(1) specific, stating that "pyruvic acid is easily decarboxylated
with warm dilute sulphuric acid" and suggests that the reac-
tion proceeds via a carhene (eq 5),
(2)
(3)
As will be shown subsequently available evidence does not
(4)
support these statements. The object of this article is to
rectify the lack of information concerning the nonenzymic
decarhoxylation of a-keto acids found in many textbooks
and to correct errors in others.
Nonoxidative Decarboxylatlon
Thermal Decarboxylation
Laneenbeck and Hutschenreuter ( 5 ) found that heatine
pyruvic acid under an atmosphwr of nitrogen for 15 min a t
13: O C ~ r o d u c e dno acetaldehvde lai the u-nitronhmvlhv-
drazon;). The pyrolysis of hen&ylf&mic &id (phenyl&yok-
cylic acid, I, R = CsH5) under an atmosphere of carhon
dioxide or of nitrogen a t temperatures between 250 OC and
300 OC was investigated by Hurd and Raterink (61, who
found that the identifiable products were carbon monoxide
(20%),carhon dioxide (50%),benzaldehyde (-22%) and heu-
zoic acid (-55%). Phenylpyruvic acid (1, R = C6H5CH2)was
found to give carbon monoxide (-60%), carhon dioxide
(-44%), and phenylacetic acid (-35%), but no aldehyde. In a
more recent study Ahmad and Spencer (7) found that 10 a -
keto acids were not decarboxylated when each was heated in
boiling water for 25 min under nitrogen.
The conditions used in these studies would cause rapid
and complete decarhoxylation of p-keto acids (8). Indeed,
the conditions used by Hurd and Raterink would decarhox-
ylate most types of carboxylic acid, and it is clear that, in
relative terms, a-keto acids are thermally stable. This con-
clusion is reinforced by t h e observation t h a t , if a
dicarboxylic acid, such as 2-oxalopropionic acid (I, R =
HOZCCH(CHJ)),in which one carboxyl group is a and the
other 0 to a carhonyl group, is heated, i t is the 8-carhoxyl
group that is lost (9).
' Cf.,the easy decarboxylation of p-keto acids, which is discussed
in virtually every textbook of organic chemistry.
Volume 64 Number 7 July 1987 591
Decarboxylationin Acid Solotion
In 1884 Beilstein and Wiegand (10) reported that acetal-
dehyde and carhon dioxide were produced when pyruvic
acid was heated with dilute sulfuric acid. This report has
subsequently been quoted by a number of authors and pre-
sumably is the basis of Finar's statement that pyruvic acid is
easily decarboxylated. Reference to Beilstein and Wiegand's
original paper reveals, however, that the reactants were
heated in a sealed tube for 5 h a t 150 O C and that the yield of
products was not determined. Instead, the reaction mixture The free acid undergoes decarboxylation approximately 53
was made weakly alkaline and distilled; the distillate "gab
times faster than its anion ( 8 ) despite the fact that the
eine sehr starke Reaction auf Aldehyd". Because the condi- presence of a proton on the carboxylate group would he
tions used were vigorous, rather than mild, and because a expected to hinder loss of carhon dioxide. In this case it
comparatively small amount of aldehyde could give a "very appears that the proton is probably transferred to the oxy-
strong" reaction, it is clear that the results of the experiment gen of the P-carbonyl group via a low-energy, formally neu-
do not warrant the conclusion that pyruvic acid is easily tral, six-membered transition state that collapses to an enol,
decarboxylated by warm, dilute, sulfuric acid.
At the lower temperature of 100 OC many other a-keto (10).
acids are demonstrably stable in aqueous acid as evidenced
by the fact that they can be synthesized by routes that
involve acid-catalyzed hydrolysis of the immediate precur-
sor of the a-keto acid in question ( 2 , I I ) .
Mechanism of Decarboxylation
The resistance of a-keto acids to thermal decarhoxylation
is explicable in terms of the mechanism that is currently
accepted for the reaction (8, 12). Many carboxylic acids
undergo decarboxylation by unimolecular heterolysis of the
anion,

R?
:a C4 = R:- + COz
Other carhoxylic acids which are easily decarboxylated due
to the presence of unsaturation p to the carboxyl group are
The intermediate produced during the reaction is a carban- cyanoacetic acid,
ion; decarhoxylation would, therefore, be expected to be
facilitated by the presence of a group, within the side chain
of the acid, capable of stabilizing the carhanion by charge
delocalization, and this is found to be so. Thus, whereas
sodium acetate must he fused with soda lime to effect its
decarboxylation
and 2- and 4-pyridylacetic acid and other similar heterocy-
clic acids. From the fact that the last two acids are so easily
decarboxylated as to he difficult to isolate, i t is clear that
protonated nitrogen, either in the Bposition or conjugated to
CO, + HO- = HCO; it, is a very effective electron sink. The reactive species in the
case of 4-pyridylacetic acid is thought to he the zwitterion,

nitroacetic acid is rapidly and quantitatively decarboxylated

by heating an aqueous solution of the sodium salt at 80 "C,

A suitably placed carbonyl group is also very effective in

promoting decarboxlation as evidenced by the chemistry of
0-keto acids such as acetoacetic acid. In this case both the
anion and the free acid loose carhon dioxide. Decarhoxyl-
ation of the anion is thought to proceed via a resonance-
stabilized enolate,
while, in the case of the 2-pyridyl acid, the proton chelate
may he involved. As pointed out by Walsh, (13)the carhonyl
group is an a-keto acid is in a position where i t cannot
delocalize negative charge arising from decarboxylation.
The anion of an a-keto acid would generate an acyl anion 3,

592 Journal of Chemical Education


while an internally hydrogen bonded free acid would give
rise to a formally neutral intermediate akin to an ylide, 4.
Both would he high-energy species because delocalization of
change to oxygen would create a partially electron deficient
carbon atom and hence would be severely restricted, making
decarboxylation difficult. Intermediate 4 would also be ex-
pected to be produced during decarhoxylation in acidic solu-
tion (eq 5). Here, an additional harrier to reaction would he
the low basicity of oxygen.
Catalysis of Nonoxldatlve Decarboxylatlon
Catalysis by Amines (14, 15)
The extensive studies of Langenheck and his co-workers,
and of others. have shown that ~ r i m a r vamines facilitate the
~~ ~~
decarhoxylation of a-keto acids in aqueous solution, or in
anhvdrous nhenol. a t tem~eraturesbetween 50 'C and 170
"C. Simple primary amines, ahirh are poor carnlystli, proba-
hlv orodure s raitterionir Schiff hase in which the posirivrly
chaiged nitrogen forms an effective electron sink,
5
RCHO + HxNRL
5 + RCHO
Because nitrogen is considerably more basic than oxygen,
the concentration of the nitrogen ylide produced would he
considerably larger than the concentration of the analogous
oxygen ylide (4), thus accounting for the catalytic effect of
the amine. In one sense, however, the term catalytic is inap-
propriate because, regardless of whether the reaction is per-
formed in water or in phenol, the amine is quickly consumed
via side reactions. Support for the involvement of a nitrogen
ylide in the catalysis is provided by the fact that species of
this type have been trapped during the decarhoxylation of a
number of heterocyclic acids in which the carboxyl group is P
to a nitrogen atom, the Hammick reaction (81,
Primary amines that possess a p-carhonyl group were found
to he much more effective than simple amines as catalysts
'for the decarhoxylation of a-keto acids. The most active
compounds of this type are derivatives of 3-amino-2-oxin-
dole,
The parent compound is claimed to he about 650 times more
active than ethylamine. Again, catalysis is thought to be
effected by formation of a Schiff base. In these derivatives,
however, the p-carbonyl group is thought to promote a pro-
totropic shift, possibly via a cyclic transition state. The
product of the shift (8) contains an unsaturated system B to
the carhoxyl group, and it is this structural feature that
facilitates decarboxylation:
8 6 + RCHO
- RCHO
In comparison with primary amines the effect of secondary
and tertiary amines on the decarhoxylation of a-keto acids
has not been extensively investigated. Brown (16) claims
that secondary amines produce catalysis hut cites no sup-
porting evidence. It has long been known that quinoline
catalyzes the decarboxylation of many types of carboxylic
acid (17). Clark and others (18) have shown that the tertiary
amine promotes the decarhoxylation of both a- and 0-keto
acids and their anions. The results of thestudies suggest that
the base facilitates loss of carbon dioxide by formation of a
bimolecular complex with the carbon atom of the carboxyl
group of the acid (9),
Catalysis by Cyanide (14, 15)
Catalysis of the decarhoxylation of a-keto acids, and of
their conversion to acyloins, by cyanide was first investigat-
Volume 64 Number 7 July 1987 593
ed by Franzen and Fikentscher. Cyanide is a good nucleo-
~ h i l and
e readily adds to the carbonyl group
- - of the acid. The
adduct an electron s i n k i n a position 6 to the
carboxyl group and hence is susceptible to easy decarboxyl-
ation. Finally, cyanide is a good leaving group; the aldehyde
which results from loss of cyanide can react with the carban-
ion (lo), to give an acyloin, or can he trapped as the dime-
done derivative:
dimedone derivative
'V
CKCHO (i) dimedone, aq. dioxan.
10%
CHRCOCH(OH) CHJ + CN-
Catalysis by Thiamine Pyrophosphate-Dependent Enzymes
(13-15)
The studies of the nonenzymic decarboxylation of a-keto
acids reviewd in P r e ~ ~ i o t ~ ~ indicateihat n ~this type
s r ~ ~ t i ~ ~11)
ut acid is resistant tar thermal decarboxylation and (2) the
process can be faciliated by introducing & electron sink into
the acid, preferably bound to a 0 carbon by a multiple bond.%
It appears that because none of the 20 amino acid residues
commonly found in enzymes are capable of generating a
suitable electron sink. evolution has nroduced a snecific co-
enzyme, T P P , to do so. Studies with'^^^-dependent holo:
enzvmes.
" . with T P P itself. and with model thiazolium salts
suggest that the coenzyme functions in a manner analogous
to cyanide, as shown in the reaction,
R; , RS
? An interesting variation of this prlncnpie is described in Ahmad
and Spencer's paper (71These workers foundthat the oximes of 10
reoresenlarives o-keto acmdswere oecarooxvlaled to aive carbon -
triies in excellent yield when heated for 25 min in boilingwater. They
suggest that the zwitterions react as shown,
the negative change resulting from decarboxylation being acccom-
modated by the leaving group. This type of reaction may have bioiogi-
cal significance in view of the fact that, recently, both Chlorella and
spinach leaves have been shown to possess an enzyme that cata-
lyzes the production of hydrogen cyanide from glyoxylate oxime (R =
H) (2).
594 Journal of Chemical Education
The hydrogen atom a t position two in T P P has been shown
to be weakly acidic and the evidence that is available sug-
gests that the coenzyme is present in the active site of its
apoenzymes as the 2-anion (11).This anion is a good nucleo-
phile and, under the catalytic influence of the apoenzyme,
adds to the carbonyl group of the a-keto acid. The adduct
(12), which possesses an electron sink (positively charged
nitrogen) attached by a double bond to the carbon @ t othe
carhoxyl group, undergoes rapid enzymr-catalyzed derar-
b o x h t i o n . l'hr resonanre-stabilized carbanmn (13) that
results from h s s of eurhon dioxide can then have at least
three different fates depending on the specificity of the
apoenzyme (or, in cases where the reaction is catalyzed by an
enzyme complex, on the enzyme composition of the c&-
plex), viz.:
(1) The carbanion may be protonated, and the 2-anion of TPP may
then act as a leaving group, allowing the aldehydic decarboxyl-
ation product to diffuseaway from the enzyme (i, eq 6).
(2) The apoenzyme may catalyze the addition of the intermediate
carbanion to a carbanvl cram oresent in either the substrate a-
krro arid. ~healdehydir'drt~~rl;t>xyinric,n
produrr, or an aldehv-
dir second st~bsrntr;rhar is, the carhanion moy pnrticipole in
the formnrioll ofd!lv acyluin such as acelolactate ri, eq GI.
(3) The carbanion may be protonated and the protonated form may
then undergo two-electron oxidation as described in the next
section.
Oxldative Decarboxylatlon ( 13)
As evidenced by the n u m l m of papers cited in the re\,ieu.
hy Cooper. Cinos, and hleister (21 the nonenzymic oxidnttur
decarhoxylation ufwketu acids has received more attention
from researchers than has nonoxidative decarboxylation.
Several mild oxidants, including ceric sulfate, lead tetraace-
tate, potassium permanganate, peroxyphthalic acid, and hy-
drogen peroxide (the last either alone or in the presence of
base or of ferrous ions) are known to cause rapid, quantita-
tive decarboxylation of the title compounds.
 It would he interesting to relate the mechanisms of these
nonenzymic reactions with the mechanisms of correspond-
ine.. enzvme-catalvzed
, reactions as was done in the previous
iectiun for nonohidative decarhouylatiun. This, however, is
not Dossible beca11.i~ the mechanisms hy which the nonen'y-
mic oxidative reactions occur appear, in many cases, to be
complex and cannot be specified in detail.
In contrast, the mechanism by which the enzyme-cata-
lyzed oxidative decarhoxylation of a-keto acids occurs ap-
pears to he well authenticated. The two reactions (eqs 3,4)
given in the first section that involve oxidation are catalyzed 0
by similar enzyme complexes. Both contain three catalytic CoASH
components, viz., a decarboxylase, a dihydrolipoamide trans- R H~ ~ J + 11 RC0ShA +
. . and a dihvdrolivamide dehvdropenase.
acvlase. . - The TPP-
dependent decarboxylase components catalyze decarboxyl-
ation of their substrates as described in the previous section.
The carbanions (13) that are produced arethen protooated R1, R2, R3, R4.m in eq 6: R5 = (CH2)&ONHtransacylase;
and the protonated materials (14) act as the substrates of CoASH = coenzyme A
the appropriate transacylase.
The transacylases both utilize lipoic acid Literature Clted
1. Strver. L. Biachemistrv. 2nd od.: Freeman; Ssn Frsneixo. 1975: Lehnineu, A. L.
~ i o ~ h r m i s l rW&
y: New York, 1970.
2. Cooiler,A. J. L.: Cinoa, J. 2.; Meirter, A. Chom. Reus. 1983,83,321.
3. ~ a r e hJ., Orgonie Chemiaiy. 3rd ed.:Wiloy: New York. 1985; p 563.
4. Finar, I. L. Organic Chemistry, 6th ed.:Longmans: London, 1973; p 305.
5. Laneenheck. W.: Hutschenreuter.R. 2.Anorn Alleam. Chemie 1930.186.1.

7. Ahrnad, A.:Speneer.I.D.Con. J. Cham. 1961.39.1340.

as coenzyme; the cyclic disulfide is hound to the apoenzymes
by an amide bond involving the 6-amino group of a lysine
residue. In both cases the carbanion (13) is regenerated and
the transacylase catalyzes its addition to lipoic acid. Expul-
3. Brown. B. R.Quwt.Re". Chom. Soc. 1951.5,131.
9. Kubsia.G.: Marteii. A.E.J.Am. Chem Soe. 1981.103.7M9.
. .
10. Beilst& F.: wie&d,E, Ber. 1884. 17,841.
11. Water.K.L. Chem.Reua. 1947.42,585.
12. Could, E. S.Mechanism and Structure in Organic Chemistry; Halt: London, 1959: p
3"a
"7".

13. Wslsh, C . EnrymoticRmction Mechanism; heeman: San Franeirreo. 1979:p 683.

sion of the T P P anion from the adduct generates an S-
acyldihydrolipoamide and completes two-electron oxidation
of the decarboxylated a-keto acid. Finally, the acyl group is
transferred from the dihydrolipoamide to coenzyme A (eq
7). The remaining component of each enzyme complex, the
dehydrogenase, is concerned with reoxidation of reduced
lipoamide.
14. Bruiee, T. C.; Benkovie, S. Bioorgonic Mechonirms: Benjamin: Now Vork, 1966; Val.
11. Chaptor 8.
15. Bender, M . L. Meehonianu of Homogrnsous Catalysis from Protons to Pmfeim:
Wiloy: New York, 1971: Chapter 6.
16. Brown, J. M. In Comprehenaiua Organic Chamisfry; Barton. D.: Ollis, W . D.. Eda.;
Pergsmon: Oxford, 1979.
17. Fi8ser.L. F.;Fieser, M . Raogenfsfor OigonicSynfhasis: Wiley: New Y a k , 1962~975.
13. Clark. L. W.In The Chemistry of Carborylic Acids ond Esters; Patai. S..Ed.: Inters-
cience: London. 1969:p 539.

Volume 64 Number 7 July 1967 595