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Spores and Significance
Spores and Significance
• In the laboratory
– by nutrient limitation by exhaustion of nutrients during
growth or shifting from a rich to a poor medium.
– Addition of an inhibitor of guanine nucleotide (decoyinine)
• Massive sporulation takes place when cells enter stationary
phase. Stationary-phase events include:
• (i) synthesis and secretion of degradative enzymes such as
amylases and proteases
• (ii) synthesis and secretion of antibiotics such as gramicidin or
bacitracin
• (iii) in some species, synthesis and release of protein toxins
• (iv) development of motility
• (v) killing and cannibalism of sister cells in the population
Bacterial spore structure
• The Spore, Characteristics
– a dehydrated, multilayered structure
– spore is metabolically inert
– remain stable for months to years
• Contains:
– Complete copy of the chromosome
– minimum amount of essential proteins and
ribosomes
– high concentration of calcium bound to
dipicolinic acid
The Spore
• Dormancy
– Spores have no metabolic activity – due to low water content
– No enzyme action
– Long term survival due to extreme resistance to heat,
radiation, chemicals, desiccation
– Spores resistance is due to
• Low water in the core – dehydrated state
• SASP
• impermeability
Sporulation
• Morphological changes
– sporulation begins with the isolation of a terminal nucleus by
the inward growth of the cell membrane
– Enfolding of CM will lead to the formation of a double
membrane structure forming inner membrane and CW facing
outside, which engulf the developing spore
– The remaining layers of the spore will form
• Outer spore membrane and cortex will form between the
facing membranes
• The coat and exosporium will form out side the
membranes
• In the core, all vegetative cell enzymes become degraded
and replaced by spore constituents
Process of endospore formation
• DNA is replicated
-The spore septum begins to isolate the newly replicated DNA
and a small portion of the cytoplasm
• The plasma membrane starts to surround the replicated DNA
and cytoplasm
• The spore septum surrounds the isolated portion forming a
forespore
• The peptidoglycan layer forms between the 2 membranes
• Thick spore coat forms around the outside membrane, making
endospores resistant to many harsh chemicals
-The new spore loses its water and becomes highly dehydrated
to the point that all that remains in the spore is:
DNA ; small amounts RNA ; ribosomes, enzymes
• Endospore is released from the cell, can remain dormant for
1000s of years
Process of endospore formation
Spore Resistance
• Heat resistance
– Implications in industry
– Withstand 100o C for several minutes
– Dt value is used for quantification
• Dt is time in min at temp (t) to kill 90% of population
– The target of heat is not known- DNA is not damaged, no
mutations due to heat, but may be proteins
• C. botulinum spores, the most heat resistant
– Spores must be reduced by 12D
– 12D is the time required in a thermal process for a 12 log
reduction of the spores
– D value: time required for one-log reduction in viability
– one-log = 90% killing of the population
Germination
– Outgrowth
• new vegetative cell emerges followed by active biosynthesis and
terminates in cell division
• Outgrowh requires a supply of all nutrients essenial for cell
growth.
Macromolecules