Industrial Microbiology Session 5

Industrial Microbiology
Instructor: Bita Zamiri

Quick Recap
Microbial Structure and Function
• Microorganisms can be both very easy and very
difficult to work with.
• They reproduce rapidly.
• They can be grown in large population in the lab.
• They cannot be seen directly and are therefore
analyzed through indirect means such as
microscopes.
• Archaebacteria or archea (‘ancient bacteria’) are quite different from
• They are found everywhere eubacteria and have some features that are similar to eukaryotic cells.
• Most archaens live in extreme environments similar to those that early life
forms are through to have endured.
• Three basic types are found: halophiles ( adapted to high salt concentrations),
methanogens (methane producers) and thermophiles ( adapted to high
temperatures) and some of them are also barophiles ( adapted to high pressure).
• Prokaryotic cells are
normally less than 5 µm
in diameters • Eubacteria(‘true bacteria’) are a very diverse groups that may be divided into
12 subgroups.
• Most prokaryotic cells • However almost all industrial bacteria are contained within just two of
contain a single circular them: the proteobacteria and the Gram-positive eubacteria:
chromosome composed
of DNA, 1. The Proteobacteria is a major kingdom of Gram-negative bacteria.
2. The Gram-positive eubacteria

• Almost all prokaryotes
have cell wall made of
peptidoglycan.
• Cell division in
prokaryotes is normally
by simple binary fission.
2
Today we will study:
• Bacterial cell structure and function
• Microbial Growth and Nutrition

Microbial cell structure and function
• There is probably no such thing as a typical prokaryotic cell.
• There is a vast amount of diversity, including morphological diversity (size and shapes; rods, cocci, spirals, filaments,
etc), structural diversity (Gram-positive or Gram-negative cell walls/envelopes, external structures such as
flagella and pili, and the ability to form spores, along with metabolic, ecological and behavioral diversity.
• Classification of bacteria can be based on several major properties:
1. Morphology
2. Cell wall (gram and other stains)
3. Metabolic behavior (e.g. oxygen)
4. Obligate intracellular
Microbial cell structure
and function
• The Gram stain developed in 1884 by the Danish physician
Christian Gram, is the most widely employed staining procedure in
bacteriology.
• It is a differential staining procedure, because it separates the

bacteria into separate groups ( gram-positive and gram-negative)
based on staining procedures.
Step 1: stained with the basic dye crystal violet
Step 2: Treated with Iodine solution ( Iodine increases the interaction

between the cell an the dye)
Step 3: Decolorization by washing with ethanol or acetone. In this step,

gram positive bacteria retain the crystal violet , whereas gram-negative
bacteria lose it.
Step 4: Counterstaining with a simple basic dye different from crystal

violet (Safranin)
Microbial cell structure
and function
Microbial cell structure and
function
Cell Wall
• Because of the thicker peptidogylcan

(murein) layer, the walls of gram-
positive cells are stronger than those of
gram-negative bacteria.
• Although the Archea can be either

gram-positive or gram negative, their
cell walls are distinctive in structure and
chemical composition. The walls lack
peptidoglycan and are composed of
proteins, glycoproteins or
polysaccharides.
• Take a look at the cross section of the

bacterium E.coli:
Cell Wall
• Peptidoglycan or murein in an enormous polymer composed of many identical subunits.
• It contains two sugar derivative N-acetylglucosamine and N-acetylmuraic acid, and several different amino
acids, three of which- D-glutamic acids, D-alanine and meso-diaminopimelic acid
• The presence of D-amino acids protects against attack by most peptidases.

Microbial cell structure and
function
Cell Wall
• Chains of linked peptidoglycan subunits are
joined by cross-links between the peptides.
• Often the carboxyl group of the terminal D-

alanine is connected directly to the amino
groups of diaminopimelic acid, but a peptide
interbridge may be used instead.
• Most gram-negative cell wall peptidoglycan

lacks the peptide interbridge.
• There is probably no such thing as a typical prokaryotic cell.
• There is a vast amount of diversity, including morphological diversity (size and shapes; rods, cocci, spirals, filaments,
etc), structural diversity (Gram-positive or Gram-negative cell walls/envelopes, external structures such as flagella and
pili, and the ability to form spores, along with metabolic, ecological and behavioral diversity.
• Classification of bacteria can be based on several major properties:
1. Morphology
2. Cell wall (gram and other stains)
3. Metabolic behavior (e.g. oxygen)
4. Obligate intracellular
Microbial growth and Nutrition
• The biosynthesis of cellular components necessary for growth, reproduction and maintenance requires a supply of basic
nutrients and energy sources.
• Nutritional classification is established on the basis of specific sources of energy, electrons/hydrogen and carbon.
• Microbial cells must obtain a range of chemical elements. Four of these, the macronutrients carbon, hydrogen, oxygen
and nitrogen must be available in gram quantities per litre of growth medium.
• These elements along with phosphorous and sulphur are the principal components of major cellular polymers.
• Other major elements, including calcium, iron, potassium, magnesium are required at levels of a few milligrams per
liter; the trace elements primarily cobalt, copper, manganese, molybdenum, nickel, selenium and zinc are needed in
only microgram quantities.
• Microbial growth can be defined as an orderly increase in cellular components resulting in cell enlargement and
eventually leading to cell division.
• This implies that a consequence of growth is always an increase in cell numbers. However under certain conditions,
growth can occur without cell division, for examples when cells are synthesizing storage compounds e.g. glycogen.
iN this situation, the cell numbers remain constant, but the concentration of biomass continues to increase.
• This is also true for some organisms such as fungi. Their growth results only in increased size. ( We sill study this later)
Lag Phase: Virtually no growth occurs and the microbial
population remains relatively constant. However, its is a period of
intense metabolic activity as the microbial inoculum adapts to the new
environment.
• When cells are inoculated into fresh medium, they may be

deficient in certain enzymes, vitamins or cofactors and etc, that
must be synthesized in order to utilize available nutrients, prior to
cell division taking place.
• The chemical composition of the fermentation media influences

the length of the lag phase.
• It is usually longer if the inoculum was grown up using a carbon

source different from that in the fresh medium because cells must
synthesize enzymes required to catabolize the new substrate.
• Physiological stress may also have an effect especially as cells are

often transferred from an inoculum medium of low osmotic
pressure ( low solute concentration) to fresh medium of higher
osmotic pressure (hig solute concentration).
• Other factors that influence the length of the lag phase: age,
concentration, viability and morphology of the inoculum.
• Usually inocula prepared from cells harvested in the exponential

growth phase exhibit shorter lag phases than those harvested from
subsequent stages.

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Industrial Microbiology

Instructor: Bita Zamiri

2. The Gram-positive eubacteria

• Bacterial cell structure and function

• Microbial Growth and Nutrition

• Classification of bacteria can be based on several major properties:

2. Cell wall (gram and other stains)

3. Metabolic behavior (e.g. oxygen)

• It is a differential staining procedure, because it separates the

Step 1: stained with the basic dye crystal violet

Step 2: Treated with Iodine solution ( Iodine increases the interaction

Step 3: Decolorization by washing with ethanol or acetone. In this step,

Step 4: Counterstaining with a simple basic dye different from crystal

• Because of the thicker peptidogylcan

• Although the Archea can be either

• Take a look at the cross section of the

• The presence of D-amino acids protects against attack by most peptidases.

• Often the carboxyl group of the terminal D-

• Most gram-negative cell wall peptidoglycan

• Classification of bacteria can be based on several major properties:

2. Cell wall (gram and other stains)

3. Metabolic behavior (e.g. oxygen)

• When cells are inoculated into fresh medium, they may be

• The chemical composition of the fermentation media influences

• It is usually longer if the inoculum was grown up using a carbon

• Physiological stress may also have an effect especially as cells are

• Usually inocula prepared from cells harvested in the exponential

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