Module 2.

BACTERIAL GENETICS,
METABOLISM AND MORPHOLOGY
MLS 2309 | CLINICAL BACTERIOLOGY
College of Medical Laboratory Science | Central Philippine University
 LEARNING OUTCOMES
REFERENCE At the end of this module, the learner should
BAILEY AND SCOTT’S have been be able to:
DIAGNOSTIC
MICROBIOLOGY 1
Enumerate correctly the general
characteristics of bacteria

Discuss comprehensively the concepts of

CHAPTER 2 2 bacterial genetics and its importance in the
Bacterial study of bacteria.
Genetics,
Metabolism, 3 Explain comprehensively the processes of
bacterial metabolism.
and Structure
Describe accurately microscopic morphology
4 (i.e., size, shape, cell-to-cell arrangement,
staining reaction)
 CHARACTERISTICS OF BACTERIA

Prokaryotes

Minute Reproduce
0.25-1 um
width, 1-3 um
by binary
length fission
Possess
Unicellular both DNA
and RNA
 BACTERIA
• Plural of bacterium
• First Known Use: 1881

Group of microscopic, single-

celled organisms that inhabit
virtually all environments,
including soil, water, organic
matter, and the bodies of
multicellular animals. Though
some bacteria can cause food
poisoning and infectious diseases
in humans, most are harmless
Needle (purple)
and many are beneficial.
Merriam-Webster Dictionary
contaminated with
bacteria (yellow).
 BACTERIAL GENETICS

• Cell’s aim is to produce

proteins responsible for
cellular structure and
function
• Bacterial DNA encodes
genetic information
• Bacterial RNA acts as a
blueprint for protein
construction
 Bacterial
Replication
• Bacteria replicate by binary
fission

• Binary fission begins when

bacterium DNA divides into
two (replicates)
• Binary fission faster means of
cellular division than mitosis
Salmonella undergoing binary fission.
The cell divides resulting in the formation of
two identical cells. (Janice Haney Carr/CDC)
 Chapter 2 Bacterial Genetics, Metabolism, and Structure 9

Process of Binary Fission

3’ 5’

Replica DNA Replica

A A T

A
5’ 3’
polymerase

T
A
T

T
A

T
C
T
T

A C
G
C G A

C T
A C
G C
G T
C

G
C
C
Replication fork

G
A

C
Origin of

G
G
C
replication

C
A
G
T

T
Daughter Daughter

T
5’

A
A

C
3’ 3’ 5’ 3’ 5’ strand strand

G
3’ 5’
3’
5’ 5’
5’ 3’ 5’ 3’
3’ T A
C G
Bidirectional replication G C

A T
A T
T A
G C

C G
G C
A T
3’ 5’

Parent
strands

Terminus

Figure 2-4 Bacterial DNA replication depicting bidirectional movement of two replication forks from origin of replication. Each parent

Single DNA
strand serves as a template for production of a complementary daughter strand and, eventually, two identical chromosomes.

Bacterial DNA replication

chromosomes, each containing two complementary
strands, one of parental origin and one newly syn-
thesized daughter strand. Although the time required
enzymes and cofactors, and RNA molecules of specific
structure and function.
molecule
depicting bidirectional
for replication can vary among bacteria, the process
generally takes approximately 40 minutes in rapidly
growing bacteria such as E. coli. However, the replica-
Transcription. Gene expression begins with
transcription, which converts the DNA base sequence
of the gene (i.e., the genetic code) into an mRNA replicates and Cell wall forms
movement of two replication
tion time for a particular bacterial strain can vary (messenger RNA) molecule that is complementary to
depending on environmental conditions such as the
availability of nutrients or the presence of toxic sub-
the gene’s DNA sequence (Figure 2-5). Usually only
one of the two DNA strands (the sense strand) encodes both copies between the two
forks from origin of replication. attach to the cell
stances (e.g., antimicrobial agents). for a functional gene product, and this same strand is

Expression of Genetic Information

the template for mRNA synthesis.
RNA polymerase is the enzyme central to the DNA molecules
Gene expression is the processing of information en- transcription process. The enzyme is composed of four
membrane. Cell membrane dividing the original
Each parent strand serves as
coded in genetic elements (i.e., chromosomes, plasmids, protein subunits (α [two copies], β, β′) and a sigma (σ)
and transposons) that results in the production of factor. Sigma factor is loosely affiliated with the
biochemical products. The overall process is composed enzyme structure and identifies the appropriate site
grow between the cell into two
a template for production of
of two complex steps, transcription and translation, and on the DNA template where transcription of mRNA is

two DNA
requires various components, including a DNA template initiated. This initiation site is also known as the

identical daughter
representing a single gene or cluster of genes, various promoter sequence. The remainder of the enzyme

a complementary daughter molecules. cells (cytokinesis).

strand and, eventually, two
identical chromosomes.
Mutation
May be spontaneous or induced
Genetic Change
(mutagens)
in Bacteria
Genetic Recombination
Also homologous recombination
Basic Mechanisms
Genes are transferred between
homologous regions on two DNA
molecules
Chapter 2 Bacterial Genetics, Metabolism, and Structure 15

Gene Exchange A Recombination

Includes transformation,
Rec A protein

transduction, and conjugation.

Recipient Uptake of donor Alignment of donor DNA with Recombined DNA
DNA ("foreign") DNA homologous recipient DNA fragment (blue)

GENETIC ALTERATIONS
B Transformation

DNA sequences that code for the amino acid Donor

Free DNA
Recipient

sequence in one protein may be sliced up and/or

combined with other polypeptides. Cell lysis and release of free DNA Uptake and recombination
C Transduction
 Chapter 2 Bacterial Genetics, Metabolism, and Structure
Rec A protein

DNA Exchange
Recipient Uptake of donor Alignment of donor DNA with Recombined DNA
A Recombination
DNA ("foreign") DNA homologous recipient DNA fragment (blue)
Rec A protein

B Transformation

in Bacteria
Recipient
Donor Uptake of donor Alignment of donor DNA with Recombined
Recipient DNA
DNA ("foreign") DNA homologous recipient DNA fragment (blue)
Free DNA
Chapter 2 B

B Transformation
Recipient Uptake of donor Alignment of donor DNA with Recombined DNA
DNA
Donor ("foreign") DNA homologous recipient DNA A Recombination Recipient
fragment (blue)
Rec A protein
Cell lysis and release of free DNA Uptake and recombination
Free DNA
C Transduction

TRANSFORMATION
B Transformation
Donor Recipient Uptake of donor
Recipient
Alignment of donor DNA with
DNA ("foreign") DNA homologous recipient DNA

Free DNA
Cell lysis and release of free DNA Uptake and recombination
Bacterium binds environmental DNA and
B Transformation
C Transduction Donor

transports it across the cell membrane.

Donor cell DNA Release of bacteriophage Bacteriophage infects and Free DNA

packaged in bacteriophage from donor cell releases donor DNA

Cell lysis and release of free DNA Uptake and recombination
Cell lysis and release of free DNA

D Transduction
C Conjugation: Chromosome transfer C Transduction

Donor Recipient

TRANSDUCTION
Donor cell DNA Release of bacteriophage Bacteriophage infects and
packaged in bacteriophage from donor cell releases donor DNA
Donor cell DNA Release of bacteriophage
packaged in bacteriophage from donor cell

D Conjugation: Chromosome transfer

Involves bacteriophages
D Conjugation: Chromosome transfer
Donor
Donorcell DNA Release of bacteriophage
Recipient Donor Bacteriophage infects
Recipient and

packaged Transfer
in bacteriophage
of newly synthesized chromosomal DNA from donor cell releases donor DNA
mobilized through intercellular bridge

D Conjugation: Chromosome transfer Transfer of newly synthesized chromosomal DNA

E Conjugation: Plasmid transfer mobilized through intercellular bridge

CONJUGATION
Donor Recipient E Conjugation: Plasmid transfer
Donor Recipient Recipient
Transfer of newly synthesized chromosomal DNA Donor

mobilized through intercellular bridge

Genes transferred from one bacterium to E Conjugation: Plasmid transfer Chromosome Plasmid

the other through the pilus.

Transfer of newly synthesized plasmid
Donor
Transfer of newly synthesized chromosomal Recipient
DNA DNA through intercellular bridge

Chromosome mobilized
Plasmidthrough intercellular bridge
Figure 2-8 Genetic recombination (A). The mechanisms of gene exchange between bacter
conjugational transfer of chromosomal (D) and plasmid (E) DNA.

E Transfer
Conjugation: Plasmid of newly synthesized plasmid
transfer
DNA through intercellular bridge Gene Exchange DNA include tra
As just mentioned, an organism’s opportunity for jugation.
Donor Recipient undergoing recombination depends on the acqui-
sition of “foreign” DNA from a donor cell. The three Transformat
Figure 2-8 Genetic recombination (A). The mechanisms of gene exchange between bacteria:
mechanisms transformation
by which (B),
bacteria physically transduction
exchange (C),ofan
cell uptake fre
Bacteriophages
Viruses that infects bacteria and capable
of injecting its genes (DNA or RNA).

Pilus
It is used by bacteria to transfer genetic
material between the two cells.
Bacterial conjugation is often regarded as
the bacterial equivalent of sexual
reproduction.

Plasmids
Extrachromosomal, double-stranded DNA.
They are passed to daughter cells or
transferred through conjugation. Genes
that code for antibiotic resistance are
often located on plasmids.
 BACTERIAL METABOLISM

FUELING BIOSYNTHESIS POLYMERIZATION ASSEMBLY

Acquisition of Use precursor Polymerize Assembly of

nutrients from products to building blocks macromole-
the produce into macro- cules into the
environment, building blocks molecules component
production of (amino acids, (lipids, LPS, structures of
precursor fatty acids, polysaccha- the bacterial
metabolites, sugars, and rides, proteins, cell.
and energy nucleotides). and nucleic
production. acids).
 Flagella
Murein • α—Ketoglutarate
Nutrients Amino acids
• Succinyl CoA
Pili

Protein

Overview of Bacterial Metabolism

• Oxaloacetate
Cytosol

RNA
Poly-
Nucleotides ribosomes

DNA
Fueling
Nucleoid
Biosynthetic Polymer- Assembly
Fueling products Building blocks reactions
Macromolecules Structures
reactions izations reactions

Figure 2-11 Overview of bacterial metabolism, which Lipid Inclusions

Nutrients Precursor
includes the processes of fueling, biosynthesis, polymerization,
and assembly. (Modified from Niedhardt FC, Ingraham JL,
• Gases metabolites
Schaechter M, editors: Physiology of the bacterial cell: a molecular
Carbon dioxide (CO2) approach, Sunderland, Mass, 1990, Sinauer Associates.) Fatty acids
Oxygen (O2)
Ammonia (NH3)
• Organic compounds, including amino acids
• Water (H2O)
Lipopoly-
• Nitrate (NO3-)
• Phosphate (PO43-)
saccharide
• Hydrogen sulfide (H2S)
• Sulfate (SO42-)
• Potassium (K+)
• Magnesium (Mg2+)
• Calcium (Ca2+)
• Sodium (Na+) Metabolic Sugars Glycogen Envelope
• Iron (Fe3+) Glucose
Organic iron complexes energy
Flagella
Murein
Precursor
metabolites
Nutrients Amino acids Pili
• Glucose 6-phosphate
• Fructose 6-phosphate Protein
• Pentose 5-phosphate
• Erythrose 4-phosphate
• 3-Phosphoglycerate
Cytosol
• Phosphoenolpyruvate
• Pyruvate RNA
• Acetyl CoA
• α—Ketoglutarate Poly-
• Succinyl CoA Nucleotides ribosomes
• Oxaloacetate

DNA Nucleoid
Fueling Biosynthetic Polymer- Assembly
reactions reactions izations reactions

Lipid Inclusions
Precursor
metabolites
Fatty acids
Fueling products Building blocks Macromolecules Structures
Lipopoly-
saccharide
Figure 2-11 Overview of bacterial metabolism, which
Nutrients includes the processes of fueling, biosynthesis, polymerization,
Metabolic Sugars Glycogen Envelope
 Precursor

Fueling metabolites
produced by
three central
pathways:
Acquisition of EMP pathway, To produce
nutrients (e.g., TCA cycle, energy from
diffusion, and PPP shunt. glucose,
active microorganisms
transport, 2 use 2 general
group processes:
translocation) Respiration and
Fermentation

1
3
20 Part I BASIC MEDICAL MICROBIOLOGY

Glucose

P NADPH2
Overview diagram of the
Glucose 6-phosphate 6-Phosphogluconolactone 6-Phosphogluconate central metabolic
NADPH2
pathways (EMP, TCA
cycle, and pentose
Fructose 6-phosphate Pentose 5-phosphate*

Fructose 1,6-diphosphate
Erythrose 4-phosphate
phosphate shunt)
Pentose phosphate cycle

Triose 3-phosphate • Precursor metabolites that are

FADH2
produced are highlighted in
NADH2
red.
1,3-Diphosphoglycerate
• Production of energy in the
form of ATP (~P) by substrate-
Fumarate Succinate
P
P
3-Phosphoglycerate level phosphorylation is
P highlighted in yellow.
Succinyl CoA
Reduced carrier molecules for
Malate
2-Phosphoglycerate TCA cycle •
NADH2
P
NADH2
transport of electrons used in
Phosphoenolpyruvate
Oxaloacetate
α—Ketoglutarate oxidative phosphorylation are
P NADH2 NADPH2 highlighted in green.
Acetyl
PYRUVATE CoA Citrate Isocitrate

EMP Pathway
Figure 2-12 Overview diagram of the central metabolic pathways (Embden-Meyerhof-Parnas [EMP], the tricarboxylic acid [TCA]
cycle, and the pentose phosphate shunt). Precursor metabolites (see also Figure 2-11) that are produced are highlighted in red;
production of energy in the form of ATP (~P) by substrate-level phosphorylation is highlighted in yellow, and reduced carrier molecules
Energy Production
RESPIRATION FERMENTATION
Involves O2, gylcolysis (EMP), and Does not require O2 and Kreb’s
Kreb’s cycle cycle
Glucose completely broken down Energy is released from sugars or
and results in high energy other organic molecules (e.g.,
production amino acids, purines)
In the presence of O2, glucose is Produces various end products
changed into CO2 and H2O (alcohols, acids, CO2, and H).
Carried by obligate aerobes and Carried by both obligate and
facultative anaerobes facultative anaerobes
Energy Utilization
Maintenance of
Biosynthesis of Activity of the
the physical and
new cell locomotor
chemical integrity
components organelles
of the cell

Transport of
solutes across Heat production
membranes
 BACTERIAL MORPHOLOGY

PARAMETERS
1. Size
2. Microscopic Shape
3. Cell-to-cell
Arrangement
4. Staining Reaction
GRAM STAIN
Gram-positive bacteria: blue/purple
Gram-negative bacteria: red/pink
Fundamental
Shapes
1. BACILLI – rods
2. COCCI – spherical
or round
3. SPIRILLA – spiraled;
comma shaped
• Coccobacilli – ovoid
Arrangement
of bacteria
affected by
two factors
1. Plane of division
2. Position taken
after cell
division
Arrangement
1. Singly 6. Groups of eight
• Cuboidal
2. Pairs • e.g. Sarcinae
• Diplococci
• Diplobacilli 7. Palisades
• Side by side
3. Chains • e.g. Corynebacterium
• Streptococci
• Streptobacilli 8. Chinese character
• Snapping
4. Grape-like clusters • Organisms bend at point of
• Staphylococci division
5. Groups of four
• Tetrads
• e.g. Peptococcus
 1 2 3 4

5 6 7 8

Describe the microscopic morphology.

THANK YOU!