BACTERIAL METABOLISM

Metabolism Processes

1. FUELING
Acquisition of Nutrients:
Nutrients must cross the bacterial cell envelope first.
a. Simple diffusion – nonselective uptake of substances by the bacterial cell
envelope. Does not require energy for transport. Transport mechanism of
some key nutrients (water, oxygen, and carbon dioxide).
b. Active transport – most common method used for the uptake of nutrients.
Requires energy for transport. Involves carrier molecules (embedded in the
membrane portion of the cell envelope) that combine with the nutrients,
transport them across the membrane, and release them within the cell.
(Certain sugars, most amino acids, organic acids, and many inorganic ions).
c. Group translocation – requires energy but differs from active transport in
that the nutrient being transported undergoes chemical modification. Many
sugars, purines, pyrimidines, and fatty acids are transported by this
mechanism.

Production of Precursor Metabolites

Once inside the cell, many nutrients serve as the raw materials from which
precursor metabolites for subsequent biosynthetic processes are produced.
These metabolites are produced by three central pathways:
a. Embden-Meyerhof-Parnas (EMP) pathway
b. Tricarboxylic acid (TCA) cycle
c. Pentose phosphate shunt

Energy Production
Accomplished by the breakdown of chemical substances through the degradative
process of catabolism coupled with oxidation-reduction reactions.
The energy source molecule is oxidized as it donates electrons to an electron-
acceptor molecule, which is then reduced.
Carrier molecules (mediate electron transfer):
a. Nicotinamide-adenine-dinucleotide (NAD+).
b. Nicotinamide-adenine-dinucleotide phosphate (NADP+).

The energy released by the oxidation-reduction reaction is transferred to

phosphate-containing compounds (e.g., ATP) where high-energy phosphate
bonds are formed.
General mechanisms for ATP production in bacterial cells:
a. Substrate-level phosphorylation - High-energy phosphate bonds
produced by the central pathways are donated to adenosine
diphosphate (ADP) to form ATP.
b. Oxidative phosphorylation - Involves an electron transport system
that conducts a series of electron transfers from reduced carrier
molecules (NADH2 and NADPH2) to a terminal electron acceptor.
 2. BIOSYNTHESIS
The fueling reactions essentially bring together all the raw materials needed to
initiate and maintain all other cellular processes.
The production of precursors and energy is mostly accomplished by catabolic
processes that involve the degradation of substrate molecules.
The three remaining primary pathways – biosynthesis, polymerization, and
assembly – depend on anabolic metabolism, in which precursor compounds are
put together for creating larger molecules (polymers) needed for assembly of
cellular structures.
Biosynthetic processes use the 12 precursor products in dozens of pathways to
produce nearly 100 different building blocks (amino acids, sugars, fatty acids,
nucleotides).

3. POLYMERIZATION AND ASSEMBLY

Various anabolic reactions assemble (polymerize) the building blocks into
macromolecules (lipids, lipopolysaccharides, polysaccharides, proteins, and
nucleic acids). This synthesis of macromolecules is driven by energy and
enzymatic activity within the cell.
Similarly, energy and enzymatic activities also drive the assembly of these
various macromolecules into the component structures of the bacterial cell. That
is, the cellular structures are the product of all the genetic and metabolic
processes.

Mechanisms of Energy Production from Catabolized Carbohydrates

1. FERMENTATION
Anaerobic process carried out by both obligate and facultative anaerobes.
Electron acceptor: Organic compound
Less efficient in energy generation than respiration (oxidation) – the initial
substrate is not completely reduced; therefore, all the energy in the substrate is
not released.
When fermentation occurs, a mixture of end products (e.g., lactate, butyrate,
ethanol, and acetoin) accumulates in the medium.

2. RESPIRATION (OXIDATION)
Aerobic process carried out by obligate aerobes and facultative anaerobes.
Final Electron Bacterial Groups
Acceptor
Oxygen Obligate aerobes
Facultative anaerobes
Inorganic form of oxygen Anaerobes
(e.g., nitrate, sulfate)

Biochemical Pathways from Glucose to Pyruvic Acid

Glucose – starting carbohydrate for bacterial fermentations or oxidations.

 When bacteria use other sugars as a carbon source, they first convert the sugar to
glucose, which is processed by one of three pathways. These pathways are designed to
generate pyruvic acid, a key three-carbon intermediate.
Pyruvate can be further processed either fermentatively or oxidatively.

Major Metabolic Pathways Used by Bacteria to Break Down Glucose to Pyruvic Acid
Metabolic Pathway Key Characteristics
Embden-Meyerhof-Parnas (EMP) a. Major pathway in conversion of
Glycolytic Pathway glucose to pyruvate.
b. Generates reducing power in the form
NADH2.
c. Generates energy in the form of ATP.
d. Anaerobic, does not require oxygen.
Pentose Phosphate (Phosphogluconate) a. Alternative to EMP pathway for
Pathway carbohydrate metabolism.
b. Conversion of glucose to ribulose-
5phosphate, which is rearranged into
other 3-, 4-, 5-, 6-, and 7-carbon
sugars.
c. Provides pentoses for nucleotide
synthesis.
d. Produces glyceraldehyde-3-
phosphate, which can be converted to
pyruvate.
e. Generates NADPH, which provides
reducing power for biosynthetic
reactions.
f. May be used to generate ATP (yield is
less than with EMP pathway).
Entner-Doudoroff Pathway a. Converts glucose-6-phosphate (rather
than glucose) to pyruvate and
glyceraldehyde phosphate, which can
be funneled into other pathways.
b. Generates one NADPH per molecule
of glucose but uses one ATP.
NADH2, nicotinamide adenine dinucleotide dehydrogenase
NADPH, nicotinamide adenine dinucleotide phosphate

Carbohydrate Utilization
The ability of microorganisms to use carbohydrates for growth is an integral part of
most diagnostic identification schemes.
Fermentation of the sugar – detected by acid production and a concomitant change of
color resulting from a pH indicator present in the culture medium.
Bacteria generally ferment glucose preferentially over other sugars, so glucose must not
be present if the ability to ferment another sugar is being tested.

Lactose Fermentation
Lactose – disaccharide consisting of 1 glucose molecule and 1 galactose molecule
linked together by a galactoside bond.
Steps involved in bacterial fermentation of lactose:
a. First step: Requires β-galactoside permease for the transport of lactose
across the cell wall into the bacterial cytoplasm.
b. Second step: Occurs inside the cell and requires β-galactosidase to break
the galactoside bond, releasing glucose, which can be fermented. (All
organisms that can ferment lactose can also ferment glucose.)