TCA CYCLE and its ANAPLEROTIC SEQUENCES

• 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 Since intermediates withdrawn from the cycle cannot be used to regenerate OAA,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 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).
1 | TCA 2008
Dr. Shiva C. Aithal, Dept. of Microbiology, Dnyanopasak College, PARBHANI shiva.aithal@rediffmail.com
• 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 (and hence OAA) can be synthesized from
acetyl-CoA by direct reductive carboxylation catalysed by a ferredoxin-dependent pyruvate
synthase.

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