TCA CYCLE

• TCAcycle (tricarboxylic acid cycle; citric acid cycle; Krebs cycle) A cyclic
sequence of reactions which plays a central role in the metabolism of
many microorganisms, both eukaryotic and prokaryotic, as well as of
higher organisms. In eukaryotes the enzymes of the TCA cycle are located
in the MITOCHONDRION; in both prokaryotes and eukaryotes most of the
enzymes are soluble in the cytoplasm or matrix, respectively, but the
succinate dehydrogenase is membrane-bound in both cases.
• The TCA cycle can have both catabolic and anabolic functions, the relative
importance of which often depends largely on the physiological state of
the cell. When the TCA cycle is functioning catabolically, acetyl- CoA –
derived from carbohydrate metabolism or e.g. from fatty acid degradation
– enters the cycle by condensation with oxaloacetate (OAA) to form
citrate.
• With one complete turn of the cycle, 2 molecules of CO2 are eliminated, 3
molecules of NAD(P)+ and one of FAD are reduced, one molecule of ATP or
GTP is synthesized by SUBSTRATE-LEVEL PHOSPHORYLATION, and one
molecule of OAA is regenerated.
• Thus, the acetyl group is effectively completely oxidized. The NADH and
FADH2 may be reoxidized via a respiratory chain to yield energy. The
enzyme isocitrate dehydrogenase is specific for NADP in most bacteria
and e.g. in yeasts, and this reaction – together with the HEXOSE
MONOPHOSPHATEPATHWAY–is probably an important source of NADPH for
biosynthesis in many organisms.
• When functioning anabolically, the TCA cycle provides precursors for
various biosynthetic pathways. Since intermediates withdrawn from the
cycle cannot be used to regenerate OAA, OAA must be generated in some
other way if the cycle is to operate continuously.
• Various reactions for achieving this occur in microorganisms: e.g., OAA
may be formed directly by the carboxylation of pyruvate or of
phosphoenolpyruvate (PEP). Such replenishing reactions are known as
ANAPLEROTIC SEQUENCES.
• Many bacteria can use TCA cycle intermediates (di- and tricarboxylic
acids) as sole sources of carbon and energy. However, possession of the
TCA cycle does not necessarily enable an organism to grow on such
compounds; the presence of a TRANSPORT SYSTEM for the substrate is
also necessary.
• Citrate, for example, can be converted to OAA via reactions of the TCA
cycle; however, acetate must then be formed (from another molecule of
citrate) if the cycle is to continue. This may be achieved by the
decarboxylation of malate (by the ‘malic enzyme’) to pyruvate which, in
turn, is converted to acetyl-CoA by the pyruvate dehydrogenase complex.
• If acetate (or an acetate-generating substrate such as a fatty acid) is used
as the sole source of carbon and energy under aerobic conditions, acetate
can be converted to acetyl-CoA by acetyl-CoA synthetase with
simultaneous formation of AMP and PPi from ATP and can thus be
metabolized via the TCA cycle. However, if intermediates are withdrawn
1 | TCA 2008
Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI
shiva.aithal@rediffmail.com
 from the cycle for biosynthesis, OAA cannot be generated by
carboxylation of pyruvate or PEP since in aerobic organisms pyruvate
cannot be synthesized directly from acetate. Under these conditions
dicarboxylic acids are synthesized by the GLYOXYLATE CYCLE (glyoxylate
shunt) involving two reactions, catalysed by isocitrate lyase (ICL) and
malate synthase, which bypass the decarboxylation reactions of the TCA
cycle.
• Thus, in effect, one molecule of a 4-C dicarboxylic acid is generated from
two of acetate for each turn of the glyoxylate cycle; pyruvate – required
e.g. for GLUCONEOGENESIS – can be formed by the decarboxylation of
OAA generated in this way. The key enzyme in controlling whether
isocitrate is metabolized via the TCA cycle or via the glyoxylate cycle is
isocitrate dehydrogenase (ICDH).
• ICDH has a much higher affinity for isocitrate than does ICL. In
Escherichia coli, growth on acetate induces the synthesis of a bifunctional
enzyme, ICDH kinase–phosphatase, which catalyses the phosphorylation
and dephosphorylation of ICDH; phosphorylation inactivates ICDH,
allowing isocitrate to reach concentrations sufficient to permit its flow into
the glyoxylate cycle. The phosphatase function of ICDH kinase–
phosphatase is stimulated, and the kinase function inhibited, by various
metabolites– particularly pyruvate.
• Some bacteria can grow on glyoxylate (formed e.g. by purine
degradation), or on glyoxylate precursors such as glycolate, by using the
glyoxylate cycle. In this case acetyl-CoA is formed via 3-phosphoglycerate
produced by the tartronic semialdehyde pathway (= glycerate pathway)
acetyl- CoA may then condense with another molecule of glyoxylate to
form malate and hence OAA. TCA cycle enzymes are subject to various
control mechanisms affecting their biosynthesis and/or activity. For
example, in E. coli they are repressed by anaerobiosis.
• However, when conditions are such that some of the enzymes are
necessary for biosynthesis, the levels of those enzymes are increased
despite anaerobiosis; under such conditions 2-oxoglutarate
dehydrogenase (α-ketoglutarate dehydrogenase) remains repressed and
the pathway ceases to be cyclic, functioning in biosynthesis as two linear
pathways: an oxidative pathway from citrate to 2-oxoglutarate, and a
reductive pathway from OAA to succinyl- CoA. 2-Oxoglutarate
dehydrogenase is also repressed by anaerobiosis in many other
facultatively anaerobic organisms.
• The TCA cycle is inherently incomplete in some bacteria: e.g. in
cyanobacteria, in Gluconobacter, and in some members of the
Methylococcaceae; in at least some cyanobacteria succinyl-CoA is formed
via the glyoxylate cycle rather than via the reductive pathway from OAA.
A complete cycle is apparently involved in the oxidation of acetate by
sulphate in the obligately anaerobic sulphate-reducing bacterium
Desulfobacter postgatei, although some of the enzymes involved are
unusual in certain respects: e.g., malate and possibly succinate
dehydrogenases appear to use quinones instead of pyridine nucleotides
as coenzymes, and 2- oxoglutarate dehydrogenase is specific for a
ferredoxin instead of NAD+ as electron acceptor. In D. postgatei, pyruvate

2 | TCA 2008
Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI
shiva.aithal@rediffmail.com
 (and hence OAA) can be synthesized from acetyl-CoA by direct reductive
carboxylation catalysed by a ferredoxin-dependent pyruvate synthase.

3 | TCA 2008
Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI
shiva.aithal@rediffmail.com
 4 | TCA 2008
Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI
shiva.aithal@rediffmail.com