Microbiology (BT 13102)

UNIT IV: Microbial Nutrition

All forms of life, from microorganisms to human beings, share certain nutritional requirements for
growth and normal functioning.

Great diversity of nutritional types found among bacteria.

Nutritional Requirements
All organisms require following:

All organisms require a source of energy. Some rely on chemical compounds for their energy and
are designated as chemotrophs.

Others can utilize radiant energy (light) and are called phototroph.

Both chemotrophs and phototrophs exist among bacteria.

Source of electrons for their metabolism. Some organisms can use reduced inorganic compounds
as electron donors and are termed lithotrophs (some may be chemolithotrophs, others
photolithotrophs).

Other organisms use organic compounds as electron donors and are called organotrophs (some
are chemolithotrophs, others photoorganotrophs).

Require carbon in some form for use in synthesizing cell components. Also require at least small
amounts of CO2 .Some can use CO2 as their major, or even sole, source of carbon; such organisms are
termed autotrophic. Others require organic compounds as their carbon source and are termed
heterotrophic.

Also need nitrogen in some form for cell components. Bacteria are extremely versatile in this respect.
Unlike eukaryotes, some bacteria can use atmospheric nitrogen.

Others thrive on inorganic nitrogen compounds such as nitrates, nitrites, or ammonium salts, and still
others derive nitrogen from organic compounds such as amino acids.

Sulfur and phosphorus for cell components. Sulfur is needed for synthesis of certain amino acids
(cysteine, cystine, and methionine). Some bacteria require organic sulfur compounds, some are
capable of utilizing inorganic sulfur compounds, and some can even use elemental sulfur.

Oxygen is provided in various forms, such as water; component atoms of various nutrients; or
molecular oxygen.

Phosphorus, usually supplied in the form of phosphate, is an essential component of nucleotides,

nucleic acids, phospholipids, teichoic acids, and other compounds.
Trace Elements: All living organisms require metal ions, such as K, Ca2+ Mg2+ , and Fe2+ for normal
growth. Other metal ions are also needed but usually only at very low concentrations, such as Zn2+ ,
Cu2+ , Mn2+, Mo2+, Ni, B2+ , and Co2+ ;

Other Key Points:

Not all the biological functions of metal ions are known, but Fe 2+, Mg 2+, Zn 2+, Mo 6+ , Mn 2+, and Cu 2+,
are known to be cofactors for various enzymes. Most bacteria do not require Na', but certain marine
bacteria, cyanobacteria, and photosynthetic bacteria do require it.

For those members of the archaebacteria known as the extreme halophiles;' the requirement is
astonishing: they cannot grow with less than 12 to 15 percent NaCl! They require this high level of NaCl
for maintenance of the integrity of their cell walls and for the stability and activity of certain of their
enzymes.

Cofactors:

A non-protein chemical compound or metallic ion that is required for an enzyme's activity.
Cofactors can be considered "helper molecules" that assist in biochemical transformations. It can either
be ions, such as zinc and iron ions, or organic molecules, such NADP/ FADP vitamin-derived
molecules. Many of these cofactors will attach near the substrate binding site to facilitate the binding of
the substrate to the enzyme.

Coenzymes:

Cofactors can be divided into two types: inorganic ions and complex organic molecules called
coenzymes. Coenzymes are mostly derived from vitamins and other organic essential nutrients in small
amounts.

Coenzymes are organic molecules and quite often bind loosely to the active site of an enzyme and aid
in substrate recruitment, whereas cofactors do not bind the enzyme.

Structure and Function of Various Coenzymes

NAD/NADP (Nicotinamide adenine dinuceotide)

Flavin Mononucleotide (FMN) and Flavin Adenine Dinucleotide (FAD): Thiamine Pyrophosphate (TPP)

Pyridoxal Phosphate (PAL)

Holoenzymes are the active forms of enzymes. Enzymes that require a cofactor but are not bound by
one are called apoenzymes.

Apoenzyme is a protein part of the holoenzyme or conjugate enzyme. Coenzyme is the non-protein
organic group which binds itself to the Apoenzyme to form holoenzyme or conjugate enzyme.

Holoenzymes represent the apoenzyme bound to its necessary cofactors or prosthetic groups.
Vitamins:
All living organisms contain vitamins and vitaminlike compounds. These function either as coenzymes
for several enzymes or as the building blocks for coenzymes. Some bacteria are capable of
synthesizing their entire requirement of vitamins from other compounds in the culture medium, but
others cannot do so and will not grow unless the required vitamins are supplied preformed to them in
the medium.

All living organisms require water, and in the case of bacteria all nutrients must be in aqueous solution
before they can enter the cells.

Vitamin Function

Biotin (B7) : CO2 fixation

Cyanocobalamin (B12) Molecular rearrangement, cofactor in DNA synthesis, and

in both fatty acid and amino acid metabolism

Folic acid (B9) C – metabolism

Pantothenic acid (B5) Precursor of coenzyme A (CoA)

Pyridoxine (B6) Amino acid metabolism

Niacin (Nicotinic acid): B3: Precursor of NAD and NADP

Riboflavin (B2) Precursor of FAD and FMN

B1 (Thiamine) Aldehyde group transfer

Nutritional Types:
Bacteria can be divided into many groups on the basis of their nutritional requirements. All forms of life,
from microorganisms to human beings, share certain nutritional requirements for growth and normal
functioning.

Phototrophs : Required radiant energy of sunlight and transform it into chemical energy that is stored
in the bonds of CHO and other molecules. Eg. Green plants, Algae, Cyano Photosynthetic Bacteria
(Rhodospirillum, Chromatium)

Chemotrophs: Unable to use sun energy and rely on oxidation of chemical compound as a source of
energy.

Chemoorganotrophs : Obtain energy from oxidation of organic compounds. Most bacteria are
organotrophs and use CHO, organic acids and protein for energy sources.
Chemolithotrophs: Obtain energy from oxidation of inorganic compound H2S, H2 nitric (NO2) NH3 &
Fe2+ (Ferrous)

Winogradisky – 1800, Beggiatoa found in H2S rich habitat oxidizes H2S to elemental S and then to
SO42- (sulphate). This process is restricted to bacteria and not seen in higher life form.

Carbon – Imp constituent of cellular structure and metabolic compounds

Autotrophs: Org. that use CO2 as C source are autotrophic (Self nourishing). Autotrophs synthesize
organic substance from CO2 via Carbon dioxide fixation

Photoautotrophs :They obtain their energy from light .

Chemoautotrophs: They obtain their energy from oxidation of chemical compound.

Heterotrophs: Unable to use CO2 as C source and require preformed organic compound for carbon.

Chemoheterotrophic are organism that oxidize chemical compounds for their energy and required
organic forms of carbon.

Saprophytic; Symbiotic and Parasitic

A Nitrifying: NH3: NO2

Nitrosomonas

B Iron Bacteria: Fe2+ : Fe3+

Ferrobacillis

C Hydrogen Bacteria H2: H20

Hydrogenomonas

D : Sulphur Bacteria: H2S: S

Eg: Beggiatoa

S: H2So4: Thiobacillis thioxidans

Transport of Nutrients by Bacteria

1. Simple or Passive Diffusion
Ions or molecules present across the cytoplasmic membrane. It is the cytoplasmic membrane that
allows the passive passage of certain small molecules and actively concentrates others within the cell.

Except for water and some lipid-soluble molecules, few compounds can pass through the cytoplasmic
membrane (a lipid-protein, semipermeable cell membrane) by simple, or passive, diffusion.
 1. Simple or Passive Diffusion

•In this process solute molecules cross the

membrane as a result of a difference in
concentration of the molecules across the
External
membrane. Membrane Internal

•The difference in concentration (higher

outside the membrane than inside) governs
the rate of inward flow of the solute molecule.

•With time, this concentration gradient

diminishes until equilibrium is reached.

Solute
•In passive diffusion no substance in the
membrane interacts specifically with the
solute molecule as illustrated in Fig

2. Facilitated Diffusion

Mechanism by which substances cross the semipermeable cell membrane is facilitated diffusion. This
process is similar to passive diffusion in that the solute molecule also flows from a higher to a lower
concentration. But it is different from passive diffusion because it involves a specific protein carrier
molecule (called a porter or permease) located in the cytoplasmic membrane.

• The carrier molecule combines reversibly with the solute molecule, and the carrier-solute complex
moves between the outer and inner surfaces of the membrane, releasing one solute molecule on the
inner surface and returning to bind a new one on the outer surface. (Fig)

Facilitated diffusion
External Membrane Internal

Return to
original
Solute
confirmation

Carrier

Translocation
The entry of glycerol into bacterial cells is by facilitated diffusion. Although this mechanism of transport
is common in eukaryotic cells (e.g.. sugars enter them in this way), it is relatively rare in prokaryotic
cells.

Neither of the above two mechanisms, passive diffusion or facilitated diffusion, require metabolic
energy. Nor do they result in concentration or accumulation of solute against an electrochemical (with
ions) or osmotic (with nonelectrolytes) gradient.

But there are the two other mechanisms by which solutes cross membranes, both of which require
metabolic energy and accumulate substrates against concentration gradients. Solutes can be
concentrated within the cell several thousand times greater than outside the cell.

3. Group translocation

When the solute is altered chemically during transport. Best-studied group-translocation system is the
phosphoenolpyruvate—dependent sugar—phosphotransferase system. It is widely distributed in many
bacterial genera and mediates the translocation of many sugars and sugar derivatives.

These solutes enter the cell as sugar phosphates and are accumulated in the cell in this form.
Phosphotransferase system (PTS) sugar uptake and phosphorylation require the participation of
several soluble and membrane-bound enzymes. These proteins catalyze the transfer of the phosphoryl
group of phosphoenolpyruvate to the sugar molecule. The products formed are therefore sugar
phosphate and pyruvate; the overall reaction requires Mg2+. Specifically, a- relatively heat-stable
carrier protein (HPr) is activated first by transfer of a phosphate group from the high-energy compound
phosphoenolpyruvate (PEP) inside the cell.

PEP + HPr ⇌ Pyruvate + phospho-HPr

Enzyme I and HPr are soluble proteins and are nonspecific components of the process

At the same time, the sugar combines with enzyme II at the outer membrane surface and is transported
to the inner membrane surface. Enzyme II is specific for a particular sugar and is an integral component
of the cytoplasmic membrane. Here it combines with the phosphate group carried by the activated HPr.
The sugar-phosphate is released by enzyme II and enters the cell.

Sugar + phospho-HPr ..... sugar-phosphate + HPr

Enzyme I has been partially purified from several bacteria including Escherichia coli and Salmonella
typhimurium.

HPr has been purified to homogeneity from several bacteria. Mannitol enzyme II has been purified from
E. coil.

•
 Other known group-translocation processes include the uptake of adenine and butyrate at the
exterior surface of the cell and their conversion at the interior membrane surface to adenosine
monophosphate and butyryl-coenzyme A, respectively
Binding of a solute to a receptor site on a membrane-bound
4. Active transport carrier protein.

Translocation of the solute-carrier complex across the

Three steps of active
membrane.
transport

All solutes, including sugars, amino acids, Coupling of translocation to an energy-yielding reaction to lower
peptides, nucleosides, and ions, are taken up the affinity of the carrier protein for the solute at the inner
by cells through active transport. membrane surface so that the carrier protein will release solute
to the cell interior.

External Membrane Internal

Solute
Return to
original
confirmation

Carrier
ADP+Pi
ATP
Translocation

Chemiosmotic Theory
Several mechanisms have been proposed to explain the molecular basis of active transport of solutes
in microorganisms. The accumulated evidence suggests that active transport may also be explained by
chemiosmotic hypothesis advanced in 1961 by Peter Mitchell, a British biochemist.

In this case, energy released during the flow of electrons through the electron-transport chain or the
splitting of a phosphate group from ATP drives protons out of the cell. This generates a difference in pH
value and electric potential between the inside and the outside of the cell or across the membrane. This
proton gradient gives rise to a protonmotive force which can be used to pump the solutes into the cell.
When protons reenter the cell, the energy released on reentry drives the transport mechanism in the
cell membrane, probably by inducing a conformational change in the carrier molecule so that its affinity
for the solute is decreased and the solute is released into the cell interior.

Fig: How release of metabolic energy is coupled to and drives active transport
 Chemiosmotic theory
Mechanism of ATP synthesis
• The chemical reactions that lead to the synthesis of ATP are now well understood. But how the transfer of electrons through the
respiratory transport chain is coupled to the synthesis of ATP is not very clear.
• Several alternate hypotheses have been proposed to explain how energy released during electron transport is conserved in the form of
ATP.

•According to this theory, the flow of electrons through the system of carrier molecules releases
energy which drives positively charged hydrogen ions (H+), or protons, across the membranes of
chloroplasts, mitochondria, and bacterial cells.
•This movement of hydrogen ions result's in the acidification of the surrounding medium
and the generation of a pH gradient (a difference in pH) across the organelle or cell
membrane.
•In addition, such hydrogen-ion movements lead to the formation of an electric potential
gradient (a difference in charge) across the membrane (since an electric charge is
carried by the proton).
•In this way, energy released during the transfer of electrons through the respiratory
chain is conserved as a "protonmotive force"; the electric potential gradients are
produced by pumping hydrogen ions across the membrane,

• Following this first energy-

conservation step, when the
hydrogen ions reenter the
organelle or cell, they are
transported by the
membrane-bound enzyme
adenosine triphosphatase.
• Energy released on reentry
drives the synthesis of ATP,
the second energy-
conservation step

Fig: Mechanism of ATP synthesis. Flow of electrons through the respiratory chain drives hydrogen ions across the
membrane. This results in a high hydrogen-ion concentration outside the cell and a low concentration inside the cell. This
produces a pH and electrochemical gradient. ATP synthesis at the site of the ATPase complex (a knobbed structure on
the membrane) is driven by the release of energy when hydrogen reenters the bacterial cell.

Siderophores: Microbial Iron Chelators

Siderophores are small molecular iron chelators that are produced by microbes and whose most
notable function is to sequester iron from the host and provide this essential metal nutrient to microbes.

Aerobic organisms have had to evolve ferric iron—binding compounds in order to solubilize and take up
ferric iron. The iron-binding compounds formed by microorganisms are termed siderophores.
Aerobic or aerotolerant organisms are continually faced with the difficulty of obtaining enough iron for
growth, i.e., for biosynthesis of iron-containing enzymes such as cytochromes and catalase.

Most of the iron that is available for aerobic or aerotolerant organisms is present in the oxidized ferric
form, which is extremely insoluble. (Organisms that can grow under anaerobic conditions have less
difficulty in obtaining iron, since in the reduced environments in which anaerobic organisms occur iron is
in its reduced, or ferrous form, which is very soluble.).

These generally belong to two major classes, the phenolates and the hydroxamates. One well-known
member of the phenolate group is enterochelin.

For instance, cells of E. coil secrete enterochelin, which solubilizes polymeric ferric iron and forms
complexes with the ferric ions; the ferric-enterochelin complex is then transported into the bacterial
cells, where the complex is degraded and the iron reduced to the ferrous form.

Role in Plants: Plant growth-promoting rhizobacteria (PGPR) are a heterogeneous group of bacteria
associated with different plant tissues. They can enhance plant growth through different mechanisms,
including siderophores production, hence, contributing to the plant nutrition and protection against
phytopathogens.

Microbial Metabolism
Heterotrophic bacteria can use a variety of organic compounds as energy sources. These compounds
include carbohydrates, organic and fatty acids, and amino acids. For many microorganisms the
preferred compounds are carbohydrates, especially the 6-carbon sugar glucose.

Glycolysis:
The most common pathway of glucose catabolism is the Embden-Meyerhof pathway of glycolysis
(“splitting of sugar"). This process occurs very widely and is found in microorganisms. Glycolysis does
not require the presence of oxygen and therefore can occur in both aerobic and anaerobic cells.

Aerobic cells degrade glucose by glycolysis, and this process constitutes the preparatory stage for the
aerobic phase of glucose oxidation. And under anaerobic conditions this situation prevails: fermentation
 Under anaerobic conditions

fermentation
Glucose Fermentation products

Glucose fermentation
products

Under aerobic conditions

Fermentation Respiration
Glucose Intermediate 6CO2 + 6H20
(Pyruvate) O2

• In glycolysis, fructose-1,6-diphosphate formed from glucose is split into two

-1ATP
3-carbon units (dihydroxyacetone phosphate and glyceraldehyde-3-
phosphate), and they are subsequently oxidized to pyruvic acid.
• At the step where glyceraldehyde-3-phosphate is oxidized, a pair of
-1ATP
electrons (two hydrogen atoms) is removed.
• In the absence of oxygen, this pair of electrons may be used to reduce
pyruvic acid to lactic acid or ethanol. In the presence of Oxygen, this pair of
electrons may enter the respiratory chain. GAPDH
4-2=2 ATP net dehydrogenase
• Many of the reactions of the glycolytic pathway are freely reversible and can gain +2NADH
this pair of electrons may be used to reduce pyruvic acid to lactic acid or ethanol
be used for the synthesis of glucose as well as for its breakdown.
• Only three of the reactions are not reversible by common enzymes, but
the presence of other enzymes can reverse them for glucose synthesis Phosphoglycerate
+2ATP kinase
to occur.
• Thus phosphoenolpyruvate is synthesized from pyruvate by the action of
phosphoenolpyruvate synthase and specific phosphatases hydrolyze
fructose-1,6-diphosphate and glucose-6-phosphate in the biosynthetic +2ATP
direction.
• The enzymes at these steps in the degradative direction are kinases and
4
require ATP.

For each molecule of glucose metabolized, two molecules of ATP are used up and four molecules of
ATP are formed. Therefore for each molecule of glucose metabolized by glycolysis, there is a net yield
of two ATP molecules.

The overall reaction of glycolysis can be summarized as follows:

Pentose phosphate pathway (PPP)

The pentose phosphate pathway, like the glycolytic one, is another catabolic reaction pathway that
exists in both prokaryotic and eukaryotic cells. Since it involves some reactions of the glycolytic
pathway, it has been viewed as a "shunt" of glycolysis; hence it may also be called the hexose
monophosphate shunt. Its other synonym is the phosphogluconate pathway.

Glucose can be oxidized by the pentose phosphate pathway with the liberation of electron pairs, which
may enter the respiratory chain. However, this cycle is not generally considered a major energy-yielding
pathway in most microorganisms.

It provides reducing power in the form of NADPH + H+, which is required in many biosynthetic
reactions of the cell, and it provides pentose phosphates for use nucleotide synthesis.

Although it can produce energy for the cell as an alternate pathway for the oxidation of glucose, it is
also a mechanism for obtaining energy from 5-carbon sugars.

• As seen in Fig, the PPP involves the initial phosphorylation

of glucose to form glucose-6-phosphate, the latter is oxidized
to 6-phosphogluconic acid with the simultaneous production
of NADPH.
• Decarboxylation of 6-phosphogluconic acid, together with a
yield of NADPH, produces ribulose-6-phosphate.
• Epimerization reactions yield xylulose-5-phosphate and
ribose-5-phosphate.
• These two compounds are the starting point for a series of
transketolase reactions and trans aldolase reactions.

Note: that two intermediates of glycolysis—fructose-6-

phosphate and glyceraldehyde-3-phosphate--are
generated.

7
 The Entner-Doudoroff pathway (EDP)

Another pathway of glucose catabolism

• It is found in both aerobic and anaerobic procaryotes but not
in eucaryotes.
• It is fairly widespread particularly among Gram-negative
bacteria.
• As shown in Fig, glucose is phosphorylated to glucose-6-
phosphate.
• It is then oxidized to 6-phosphogluconic acid. Dehydration
step follows to yield 2-keto-3-deoxy6-phosphogluconic acid
(KDPG); the latter is cleaved to pyruvic acid and
glyceraldehyde-3-phosphate, which is metabolized via some
Embden-Meyerhof pathway enzymes to produce a second
molecule of pyruvic acid.
• In the aerobic pseudomonads the catabolism is completed
via acetyl.-CoA and the tricarboxylic acid cycle.
8

Fermentation:
Anaerobes also produce energy by reactions called fermentations. which use organic compounds as
electron donors and acceptors.

Facultative anaerobic bacteria and obligately anaerobic bacteria employ many different kinds of
fermentations to produce energy.

E.g., lactic fermentation. Streptococcus lactis, the bacterium responsible for the normal souring of raw
milk, dissimilates glucose to lactic acid, which accumulates in the medium as the sole fermentation
product. One molecule of glucose is converted to 2 molecules of pyruvic acid with concomitant
production of two NADH + H+.

The pyruvic acid is converted to lactic acid in the following reaction: Insufficient energy for ATP
synthesis results from this reaction, however, NAD is regenerated for further use as an oxidant.

Most heterotrophic bacteria produce several end products of the types indicated in previous Fig from
glucose dissimilation, but no single species produces all these end products. However, some
anaerobes do not have a functional glycolytic-system. They may have carbohydrate fermentation
pathways that use the pentose phosphate pathway and the Entner-Doudoroff pathway. Fermentations
of noncarbohydrate substrates, such as amino acids, involve highly specific pathways.
S.No Groups with Examples of Representative Products
Some Genera

1 Lactic acid bacteria • Lactic acid only or lactic acid plus acetic
acid, formic acid, and ethyl alcohol.
a Streptcoccus
• species producing only lactic acid are
b Lactobacillus homofermentative, and those producing
lactic acid plus other compounds are
c Leuconostoc
heterofermentative

2 Propionic acid bacteria • Propionic acid plus acetic acid and

carbon dioxide
a Propionibacterium

b Veillonella

3 Coli-aerogenes-typhoid • Formic acid, acetic acid, lactic acid,

bacteria succinic acid, ethyl alcohol, carbon
dioxide, hydrogen, 2,3-butylene glycol
a Escherichia (produced in various combinations and
b Enterobocter amounts depending on genus and
species)
c Salmonella

S.No Groups with Examples of Representative Products

Some Genera

4 Acetone. butyl alcohol Butyric acid, butyl alcohol, acetone, isopropyl

bacteria alcohol, acetic acid, formic acid, ethyl alcohol,
hydrogen, and carbon dioxide (produced In
various combinations and amounts depending on
a Clostridium
species)
b Eubacterium
c Bacillus

5 Acetic acid bacteria Acetic acid, gluconic acid, kojic acid

a Acetobacter

Such designations are established on the basis of the major end products of carbohydrate
fermentation. From this table it is evident that not all microorganisms metabolize the same
substrate in exactly the same manner.

• For example, Streptococcus Iactis and

Escherichia coil both ferment glucose but by
quite different pathways of fermentation, as
shown in Fig.

Figure: Glucose is fermented by many different

bacteria and in many different ways.

(A) E. coil fermentation of glucose results in a

mixture of products

(B) Streptococcus Iactis fermentation of glucose

produces lactic acid almost exclusively

14
 Energy production by Aerobic process

Tricarboxylic Acid Cycle (TCA)

• TCA cycle is a sequence of reactions that generate energy in the form of ATP and reduced coenzyme molecules
(NADH2 and FADH2 ).
• It also performs other functions. Many intermediates in the cycle are precursors in the biosynthesis of amino acids, purines,
pyrimidines, etc.

For example, oxaloacetic acid

and a-ketoglutaric acid (amino Oxaloacetate Aspartate Methionine
acid precursors)
Lysine Threonine

Alpha keto glutaric acid Glutamate Glutamate

Arginine Proline
15

Thus the TCA cycle is an amphibolic cycle, which Regulation of the TCA cycle
CO2 focuses on the enzyme Isocitrate
dehydrogenase, which is sensitive
means that it functions not only in catabolic to feedback inhibition by high
concentrations of ATP and NADH2
(breakdown) but also in anabolic (synthesis) Malate dehydrogenase and stimulation by high
concentrations of ADP and NAD.
reactions. (Shown in Fig)

Since the breakdown of glucose by

Succinate
glycolysis yields two acetyl-CoA dehydrogenase

molecules which, can enter this cycle, the Aconitase

overall equation for the cycle, per glucose

molecule broken down, is twice the above. Succinate
dehydrogenase
Isocitrate
dehydrogenase
The overall reaction of the TCA cycle can be Succinate
thiokinase
summarized as follows:

1 round TCA 2 round TCA

Alpha KG 6NADH2
3NADH2
dehydrogenase 2FADH2
1FADH2
1ATP 2ATP
Acetyl-CoA +3H2O + 3NAD+ + FAD + ADP+ Pi 2CO2 + CoA + 3NADH2 + FADH2+ ATP 16
Since the breakdown of glucose by glycolysis yields two acetyl-CoA molecules
 • Energy yield from the aerobic breakdown of one molecule of Energy yield in Aerobic respiration
glucose when the electrons stored in the reduced coenzyme 6ATP (2NADH2) 18 ATP (6NADH2 )

molecules are fed into the electron-transport chain. 4ATP (6FADH2 )

• Electron are transferred stepwise from the coenzyme carriers to 2ATP 6ATP (2NADH2) 2ATP (2GTP)

molecular oxygen, and this transfer is coupled to the generation of

8ATP 6ATP 24ATP =36ATP

ATP by oxidative phosphorylation. Glycolysis Gateway step TCA cycle

• For each glucose molecule broken down, there are 12 reduced
coenzymes to be oxidized: 2FADH2 (1 from each turn of the TCA
cycle) and 10NADH2 (2 from glycolysis; 2 from the gateway
step between glycolysis and the TCA cycle, i.e., pyruvic acid
to acetyl-CoA; and 6 from two turns of the TCA cycle).
• Since 3ATP are produced from each NADH2 and 2 ATP from each
FADH2, there are 34 ATP generated from the reduced coenzymes
via oxidative phosphorylation through the respiratory chain.
• But the total yield of ATP from the aerobic respiration of 1
glucose molecule is 38: 34 from the oxidation of reduced
coenzymes, 2 from glycolysis, and 2 from the side reaction of
the TCA cycle, that is, from 2 GTP.

Figure: ATP yield per glucose molecule broken

17
down in aerobic respiration.

Catabolism of Lipids:

Many heterotrophic microorganisms can degrade exogenous proteins, using the products as
carbon and nitrogen energy sources.

• Since protein molecules are too large to pass into, the cell, bacteria secrete exoenzymes
called proteases that hydrolyse exogenous proteins to peptides, which are then transported
into the cell cytoplasm. AcetylCoA is a common intermediate of carbohydrate and lipid metabolism,
and the TCA cycle is the common pathway for oxidation of carbohydrates, lipids, and amino acids.

Bacteria produce peptidases that break down peptides to the individual amino acids, which are
then broken down according to the specific amino acid and the species or strain of bacteria
breaking it downWhere amino acids are broken down, the carbon skeletons of the amino acids
undergo oxidation to compounds that may enter the TCA cycle for further oxidation. Entry into
the TCA cycle can be via acetyl-CoA, a-ketoglutaric acid, succinic acid, fumaric acid, or
oxaloacetic acid.

Respiration without oxygen in some Bacteria

Some bacteria which are ordinarily aerobic can grow anaerobically if nitrate is present. for
example, Aquaspirillum itersonii, an aquatic bacterium, is dependent Bacteria on oxygen unless
potassium nitrate is added to the medium. In such cases nitrate essentially substitutes for
oxygen as the final electron acceptor in the respiratory chain. This process is termed anaerobic
respiration. The pathways for the dissimilation of the carbon and energy sources are identical
with those in aerobic respiration, and electron transport occurs via a respiratory chain similar to
 that in aerobic cells. Oxygen is replaced as the terminal electron acceptor by nitrate. However,
in some strict anaerobes, other compounds, such as carbon dioxide, or ions, such as sulfate
ion, can be the terminal electron acceptors.

The Glyoxylate Cycle

The glyoxylate cycle is used by some microorganisms when acetate is the sole carbon source or during
oxidation of primary substrates (such as higher fatty acids) that are cleaved to acetyl-CoA without the
intermediate formation of pyruvic acid.

This pathway does not occur in higher organisms because they are never forced to feed on 2-carbon
molecules alone.

Specific enzymes of the glyoxylate cycle are isocitrate lyase and malate synthase. These two enzymes
fit together with other reactions of the TCA cycle to provide a bypass around some of the TCA-cycle
reactions

Acetyl-CoA enters the cycle at two places.

1. It condenses with oxalacetate to give citrate, which is the entry point for the TCA cycle, and the
further reaction leads to the formation of isocitrate. Isocitrate lyase is a splitting enzyme that produces
succinate and glyoxylate.

2. The second acetyl CoA molecule condenses with glyoxylate to give malate by the action of malate
synthase.

• Enzymes which carry out replenishment reactions are known as anaplerotic enzymes, their
function is to maintain the pool of essential intermediates for biosynthesis.

Sources

1. Michael J. Pelczar, JR., E.C.S. Chan, Noel R. Krieg, Microbiology, Fifth Edition.

2. Joanne M. Willey, Linda M. Sherwood, Christopher J. Woolverton, Prescott,

Harley, and Klein’s Microbiology, Seventh Edition.