Influence of Temperature
T Ross and DS Nichols, University of Tasmania, Hobart, TAS, Australia
Ó 2014 Elsevier Ltd. All rights reserved.
This article is a revision of the previous edition article by T. Ross, D.S. Nichols, volume 1, pp 547–556, Ó 1999, Elsevier Ltd.
Introduction
Temperature control is perhaps the most widely used method
of manipulating the microbial ecology of foods. It can be used
to inhibit growth of spoilage or pathogenic organisms, to
inactivate or kill unwanted microorganisms, or to optimize
growth or metabolism of microorganisms in fermentations.
The patterns of the effect of temperature on the ecology of
bacteria, yeasts, and filamentous fungi are remarkably uniform.
In general,
l Although microorganisms have evolved to grow within
different temperature ranges, those preferred ranges typi-
cally span only w35–40
C for bacteria, and 25–30
C for
fungi.
l Within most of that range, an increase in temperature
increases the rate of the microbial response, whether
growth–metabolism or inactivation.
l Although microorganisms have some capacity to alter their
structure and biochemistry to moderate the effects of
temperature on their activity and metabolism, they are
unable to achieve temperature homeostasis.
This entry considers the temperature limits for microbial
growth, and the effect of temperature on microbial growth rate,
metabolic rate, and composition. The physiological basis of
those responses and their consequences for the microbial
ecology of foods are also discussed. Separate entries consider
the effects of heating and freezing on microbial populations
and physiology.
Microbial Ecology of Foods: Evolution of Specific
Microbiota
Environmental microbial ecology tends to be concerned with
open systems through which energy and chemicals flow. Food
microbiology is more often concerned with batch processes,
whether daily production runs, or the resulting individual units
of foods for retail sale. Those batches are closed systems having
finite resources of carbon and energy, and negligible capacity
for the removal of the waste products of microbial metabolism.
The features of populations growing in a batch culture are
shown in Figure 1. Under constant environmental conditions,
the pattern of population growth is ‘S’-shaped and can be
described mathematically in terms of four properties shown in
Figure 1, that is, initial inoculum level, lag time, growth rate,
and ‘maximum carrying capacity.’
The values of those properties are variable. The maximum
carrying capacity of the system is usually a property of the food.
When this level is reached, the growth of most or all groups of
organisms in the product slows greatly or ceases, a phenom-
enon that has been termed the ‘Jameson Effect.’ Thus, a slow-
growing organism or one initially present in very low numbers
602 Encyclopedia of Food Microbiology, Volume 1
may never reach that level. Conversely, if the growth rate of
organisms initially present in very low numbers is much faster
than that of all the other elements of the microbiota initially
present, then it still may achieve numerical dominance. The
steeper curve in Figure 1 is an example.
That the microbial ecology of foods often is concerned with
batch processes tends to simplify understanding of the ecology
of the system. In most foods, there are relatively few microor-
ganisms present initially, and there is little competition for
resources. Thus, bacteria and fungi present will grow at their
fastest rates possible in that environment, until the environ-
ment is either depleted of essential nutrients or until it is so
altered by the toxic metabolites of microbial growth that
growth is no longer possible. In many cases, this level is
reached when the total microbial population is of the order of
109–1010 cfu g1 or ml1 of the food product. Attainment of
maximal population densities of desirable organisms, at the
expense of other organisms potentially present, is the aim of
fermented food production and is an example of manipulation
of the microbial ecology of a food to select for the desired
fermentative microbiota. That selection is achieved by opti-
mization of the growth rate of the desired organisms in
comparison to those of other organisms. It can also be achieved
by using a high level of inoculum or a combination of both.
For an organism to contribute to the ecology of an envi-
ronment, it must be metabolically active in that environment.
That, in turn, requires that the physicochemical conditions of
the environment remain within the tolerance range of that
organism. In the current context, if temperature is beyond
the minimum or maximum tolerance of the organism, the
Long lag, fast rate Maximum
carrying
10 Short lag, slow rate
capacity
9
8
Log microbial numbers
7
6
5
Slope represents
4 relative growth rate
Initial inoculum
3
level
2
Lag time
1
0
0 5 10 15 20 25
Time (hours)
Figure 1 Microbial population growth curves, typical of growth in
foods. Microorganisms may exhibit a lag phase before the full growth
rate potential is reached. Each species of microorganism will have
a characteristic maximum growth rate, governed by its genetics and
the conditions in the food. When the total population in the food reaches
109–1010 cfu g1, the growth of most or all other components of the
microbiota will slow markedly or cease.
http://dx.doi.org/10.1016/B978-0-12-384730-0.00087-2
 ECOLOGY OF BACTERIA AND FUNGI IN FOODS j Influence of Temperature
organism will fail to thrive and eventually will be eliminated.
Temperature control can be used to slow the growth of a target
organism relative to others present and suppress its potential
effect on the environment.
Thus, knowledge of the environmental tolerance ranges of
microorganisms, and the effects of other environmental factors
on the growth rate within that tolerance range, can be used to
manipulate the microbial ecology of foods, for example, to
extend shelf life. Shelf life can be extended by promoting high
levels of desirable organisms that stabilize the microbiology of
the product, as with fermented foods, or by delaying the
attainment of high levels of undesirable organisms that would
cause spoilage of the product. The microbial ecology of foods,
however, is concerned not only with organisms that become
numerically or metabolically dominant but also equally
involves manipulating the food environment to suppress
growth of, or even eliminate, pathogenic microorganisms
whose significance bears no relationship to their contribution
to the microbial ecology of the product.
Thus, to determine the microbial ecology of a food requires
information about the types and numbers of organisms
initially present, their tolerance ranges and growth rates, the
properties of the food, and the environmental conditions that
the food was exposed to between production and consump-
tion, and the time involved. Microbial tolerance limits to fluc-
tuations of chemical and physical factors in an ecosystem do
not determine which microorganisms are present at any given
moment, but rather which microorganisms can be present on
a sustained basis in that ecosystem. The interactions, over time,
of the environment and the microorganisms present leads to
the evolution of a specific microbiota.
Frequently, temperature will be the most variable feature of
the environment of a microorganism in food. Thus, to under-
stand the effects of temperature on the microbial ecology of
foods, and to manipulate that ecology, it is necessary to
consider the effects of temperature on the rates and limits of
microbial growth.
Effect of Temperature on Microbial Growth
Growth Rate
Temperature affects the potential for microbial growth, the rate
of growth or death, the composition of the cells, and the
production of metabolites. Bacterial and fungal growth rates
respond to temperature as shown in Figure 2. There are upper
and lower limits to growth, at which the growth rate becomes
zero, and an optimum temperature at which the growth rate is
maximal. The minimum, maximum, and optimum tempera-
tures for growth are known as ‘cardinal’ temperatures. As will
be discussed later, the temperature at which growth rate is
maximal is not, of necessity, the optimum temperature for
growth.
Between the minimum and optimal temperatures, the
growth rate increases with increasing temperature. The growth
rate increase is not proportional to the temperature, but it
increases more rapidly as the temperature is increased until the
optimum temperature is neared. As the temperature increases
above the optimum, the growth rate decreases rapidly, due to
thermal inactivation of the cellular macromolecules needed for
603
3
Growth rate (generations h–1)
2 A
1
B
C
0
260 270 280 290 300 310 320 330
Temperature (K)
Figure 2 Interaction of environmental factors in determining the
growth rate of bacteria and fungi. Curves A and C represent two
organisms, each of which has adapted to a different temperature range
for growth. Curve B represents the effect of a second suboptimal
environmental factor on the growth rate of organism A. The relative
response remains the same, but the absolute growth rate is reduced at all
temperatures.
growth. At low temperatures, the growth rate does not neces-
sarily decrease indefinitely to zero, and there may be a critical
threshold temperature below which growth suddenly is not
possible. The pattern of response depicted in Figure 2 is true,
generally, for poikilothermic organisms.
Each organism has its own preferred temperature range for
growth, related to its usual growth habitat. For bacteria, the
range of growth usually spans 35–40
C, irrespective of the
preferred temperature region for growth. Fungi typically grow
over a range of 25–30
C. According to the preferred temper-
ature for growth, organisms are classified as psychrophiles,
psychrotrophs, mesophiles, or thermophiles as described in
Table 1.
Despite that organisms have adapted to different temper-
ature ranges for growth, bacteria do not exhibit complete
growth rate compensation for that preferred temperature
range. Among the fastest growing organisms known are those
that are selected for, and cause problems in, moist proteina-
ceous foods with simple sugars present. Typically, those foods
are nutritious and temperature is the only constraint to
microbial growth. Among those organisms, strains that grow
fastest at low temperatures nonetheless grow more slowly at
their optimum than those species best adapted to growth at
higher temperature. For example, the fastest known bacterial
growth rates recorded are for Clostridium perfringens, which has
a generation time of w7 min in the temperature range
40–45
C. Conversely, psychrotrophic pseudomonads, which
are the dominant species and cause of spoilage of aerobically
stored, chilled, fresh foods, have generations times at 5
C in
the range 4–5 h, Figure 3 illustrates this behavior.
Table 1 Classification of microorganisms according to
their preferred temperature range for growth
Classification Temperature at which growth rate is maximal
Psychrophile 15
C or less
Psychrotroph 25–30
C
Mesophile 35–45
C
Thermophile >45
C
604 ECOLOGY OF BACTERIA AND FUNGI IN FOODS j Influence of Temperature

8 which can take values between 1 and 0) that model the degree
of ‘nonoptimality’ of each of the other environmental condi-
7
Growth rate (generations h–1)
tions (i.e., the ‘distance’ from the respective optima).
6 Thus, the model has the general form:
5 growth rate ¼ optimum rate
4 times relative inhibition due to suboptimal
temperature (a scale from 1 to 0),
3
times relative inhibition due to suboptimal
2 water activity (a scale from 1 to 0),
times relative inhibition due to suboptimal pH
1
(a scale from 1 to 0),
0 etc.
–5 0 5 10 15 20 25 30 35 40 45 50 55
Temperature (°C) This concept can also be extended to include the inhibitory
effects of conditions beyond the optimum.
Figure 3 Comparison of the growth rates of selected foodborne There is synergism, however, when multiple inhibitory
bacteria of different thermal adaptation in nutrient-rich environments,
factors are present in the environment. That synergism is
and showing the lack of temperature compensation. The organisms
manifest in modification on the environmental limits for
depicted are among the fastest growing in their respective preferred
temperature ranges. The solid curve in the lower range (5 to 37
C) growth of microbes.
represents the growth rate of psychrotrophic spoilage pseudomonads,
the middle solid line (7–48
C) is for growth of Escherichia coli, Growth Limits
a mesophile, and the upper curve is representative of the nearly ther-
mophilic Clostridium perfringens. The dotted line is for the growth of Each organism has reasonably well-defined limits for growth in
Listeria monocytogenes and is included to show that L. monocytogenes response to individual environmental factors, when all other
is not fast growing, relative to other foodborne organisms. Nonethe- factors are optimal for growth or survival. These limits,
less, under appropriate conditions, it can multiply sufficiently to however, are altered by the other environmental conditions.
cause problems, particularly in foods of reduced water activity (e.g.,
For example, the potential temperature limits for growth are
<0.97). Under such conditions, L. monocytogenes does have a growth
reduced if a second environmental factor is at a suboptimal
rate advantage over other foodborne organisms, particularly in the
chill temperature range. level. An example of these interactions is shown in Figure 4.
The hurdle concept and, more specifically, its application
in ‘hurdle technology,’ seeks to exploit this phenomenon by
Lag Times
using combinations of levels of environmental factors, each of
For a given population, the lag time responds to temperature in which on its own is insufficient to prevent growth. When
the same way as growth rate: It often has been reported that applied simultaneously, however, these individual mild stresses
there is a direct proportionality between the lag time of a culture can interact to prevent the growth of target microorganisms.
and its generation time at any temperature. The previous
environment experienced by the cell or population, however,
7.0
also can affect the lag time. The lag time may be considered to
be determined both by the amount of work to be done to equip
the cell to adjust to a new environment, and the rate at which 6.5
that work can be done. The former component is thought to be
related to the magnitude of the change in environment, the 6.0
latter by the environment itself. In an otherwise-constant
environment, an abrupt temperature shift can induce a lag time
pH

5.5
in an exponentially growing microbial population (see section
Unification of the Microbial Response to Temperature).
5.0

Interactions with Other Factors

4.5
The patterns of microbial responses to water activity and
pH are described separately in this volume. In terms of growth 4.0
rate, those responses are superimposed on the effect of 0 5 10 15 20 25 30 35
temperature. The combined effects can be understood in terms Temperature (°C)
of the ‘Gamma concept’ sometimes applied in predictive
Figure 4 Interaction of environmental factors in determining the
microbiology. The Gamma concept proposes that the effects of
boundary between growth and death of bacteria. In the example in the
multiple inhibitory factors are additive (in terms of their rela- figure, showing the effects of temperature and pH on the growth range of
tive effects on growth rate) so that microbial growth rate can L. monocytogenes, the minimum temperature for growth depends on the
be described by an expression based on the growth rate at pH of the environment. Closed circles are conditions under which growth
the optimum conditions (i.e., optimum temperature, water was possible; crosses are conditions under which growth could not be
activity, pH, etc.) multiplied by a sequence of terms (each of demonstrated.
 ECOLOGY OF BACTERIA AND FUNGI IN FOODS j Influence of Temperature
Quantitative knowledge of the combination of levels of envi-
ronmental factors required to prevent growth is scarce. A limited
amount of data exists for a few foodborne bacteria that are
pathogenic to humans. It is believed, however, that as condi-
tions become less optimal, that is, as environmental conditions
move further away from the growth–no-growth boundary and
to the no-growth region, the rate of microbial death increases.
There are, however, minor exceptions to this pattern. For
example, if a factor other than temperature prevents microbial
growth, reduction of the temperature (i.e., moving further into
the ‘no-growth’ region) will reduce the rate of death.
An interesting observation from studies on the interaction
of environmental factors is that the optimum temperature for
tolerance to a second inhibitory factor is often at a lower
temperature than for optimum growth rate. Evidence of this
behavior can be seen in Figure 4. Suboptimal water activity due
to the presence of salt may increase the maximum temperature
at which growth is possible. This effect may be due to the
synthesis of stress proteins that provide cross-protection to
temperature stress, to the alteration of the physicochemical
structure of water due to the presence of a humectant, due to
membrane rigidification, and so on.
Interpretation of the Effect of Temperature
on Microbial Growth
Effect of Temperature on Reaction Rate
Microbial growth can be considered as a complex sequence of
chemical reactions. The chemical reactions that occur within
bacterial or fungal cells are geared toward either
l Provision of energy and reducing power from the environ-
ment to the cell (catabolism), or
l Synthesis of structural and other macromolecules required
for growth (anabolism).
The rates of those reactions, and hence of microbial growth,
are dependent on temperature as may be described by Eyring’s
absolute reaction rate equation:

energy, which often is termed the ‘activation energy.’ The
kT DGy=
V ¼  ½r  e RT [1]
h not all the reactant molecules have the same kinetic energy at
where
V ¼ rate of reaction,
k ¼ Boltzmann’s constant,
T ¼ temperature,
H ¼ Plank’s constant,
[r] ¼ concentration of reactant,
DGy ¼ Gibbs free energy of activation, or ‘activation energy,’
R ¼ gas constant.
and which is based on the Arrhenius–van’t Hoff equation.
Most metabolic reactions within cells, however, do not
occur at measurable rates without the catalytic assistance of
enzymes. Enzymes are proteins and are fundamental to all
metabolic functions. They mediate the transformation of
different forms of chemical energy.
A biochemical reaction proceeds from reactants to products
via one or more ‘transition’ states that possess a higher free
energy than that of the reactants (see Figure 5). Enough energy
605
Transition
state
Free energy
Transition
G† state
Reactant
G
Product
Reaction progress
Figure 5 An illustration of a metabolic reaction in terms of Gibbs free
energy (Gy). The change in free energy of the system (DG) determines
whether the reaction is possible. The magnitude of the free energy of
activation (DGy) influences the likelihood of the reaction and the rate. DG
and DGy are both functions of temperature.
must be supplied to the system to overcome this barrier and
allow the formation of the transition state. Thermodynami-
cally, this is quantified by the free energy of activation, DGy. The
Gibbs free energy function is derived from a combination of
the first and second laws of thermodynamics:
DG ¼ DH  TDS [2]
where
DG ¼ change in free energy of the system,
DH ¼ change in enthalpy of the system,
DS ¼ change in entropy of the system,
T ¼ temperature (K).
For a chemical reaction to occur spontaneously, the change
in free energy, DG (i.e., free energy of the products minus the
free energy of the reactants), must be negative. This require-
ment is independent of the path of the reaction (Figure 5).
Although DG indicates whether a given reaction is possible,
DGy describes the amount of energy needed to ‘drive’ the
reaction. The kinetic energy of the reactants determine
whether they have sufficient energy to overcome the Gibbs free
kinetic energy is related to the temperature of the system, but
a given temperature. Rather, the energies of the reactant
molecules form a distribution of kinetic energies, the average
of which increases with temperature. Higher temperature
increases the probability that the reactants will have sufficient
energy to overcome DGy so that the reaction can proceed to
completion. Thus, the probability of reaction, and therefore
the rate, is also dependent on temperature.
Enzymes accelerate biochemical reactions by decreasing
DGy. Decreasing DGy effectively increases the number of
substrate molecules with sufficient energy to complete the
reaction. Consequently, the reaction is perceived to occur at an
increased rate.
From eqn [1], the logarithm of rate is expected to be linearly
related to the reciprocal of temperature, with the slope of that
line being equal to the activation energy of the response. A plot
of ln(rate) vs. 1/temperature is known as an Arrhenius plot.
Figure 6 is an Arrhenius plot of the growth rate of Escherichia
coli and is typical of Arrhenius plots of microbial growth rate.
606 ECOLOGY OF BACTERIA AND FUNGI IN FOODS j Influence of Temperature

0 High-temperature by eqn [1], that is, the central ‘straight-line’ portion. At any
(inactivation)
region
temperature within the normal physiological range, the
Normal- chemical composition of the cell is essentially constant.
–1 temperature
region
Beyond this range are the high- and low-temperature regions.
Low-temperature Cells grown in the high- and low-temperature regions not only
(inactivation)
have growth rates that deviate from that predicted by eqn [1]
In(rate)

region
–2
but increasingly are different in composition to those grown in
the ‘normal’ physiological range. Transitions to the high- and
low-temperature regions have been shown to result in synthesis
–3
of proteins not expressed in the normal temperature region. As
49 °C 39 °C 35 °C 30 °C 21 °C 12.6 °C will be discussed in detail, membrane lipid composition also is
–4 altered by the synthesis and incorporation into the membrane
0.0030 0.0031 0.0032 0.0033 0.0034 0.0035 0.0036
of lipids that have the effect of maintaining membrane fluidity.
1/Temperature (K)

Figure 6 An Arrhenius plot, based on a predictive model fitted to E. coli
growth rate data, and typical of microbial growth rate responses to
temperature. The dotted line is the predicted growth rate based on data in
the ‘normal temperature region for growth.’ If growth rate data are
collected over the full biokinetic region, however, deviations from this
prediction are observed at high and low temperatures. The ‘normal
temperature region’ depicted was judged subjectively, based on the
deviation of the observed data from that predicted by extrapolation of
eqn [1], but it does correspond to temperatures beyond which measurable
changes in the composition of E. coli occur.
That plot, however, shows a deviation from the Arrhenius
relationship at high and low temperatures. This deviation often
has been attributed to the denaturation of one or several key
macromolecules required by the organism for growth, as
described in Box 1. An alternate hypothesis is that the coordi-
nation of catabolic and anabolic reactions within the cells
breaks down at high and low temperatures, leading to
a reduction in the efficiency of metabolism, and eventually to
the complete breakdown of balanced growth.
Whatever the reason, the Arrhenius plot of microbial
growth rate can be considered in terms of three regions related
to temperature. The ‘normal’ physiological range is that region
where the growth rate responds to temperature as predicted
Unification of the Microbial Response to Temperature
The previous observations and discussion offer a consistent
interpretation of the effects of temperature on microbial
growth rates and limits. The temperature limits for growth are
governed by the high- and low-temperature stability of one, or
several, key macromolecules without which growth cannot
proceed.
Growth rate increases with increased temperature in accor-
dance with eqn [1] until the increase in temperature disrupts the
conformation of enzymes, or the integration of anabolic and
catabolic rates. Thus, metabolic efficiency decreases, leading to
the observed high- and low-temperature deviations. In this
interpretation, the optimum growth temperature is viewed as
the point of equilibrium between increasing reaction rates due
to temperature and the deleterious effects of temperature on
macromolecular stability or integration of metabolism. This
interpretation also leads to an explanation of why the temper-
ature for maximum growth rate does not necessarily corre-
sponds to the temperature of maximum tolerance to a second,
suboptimal, environmental constraint, that is, the temperature
of maximum tolerance is in the mid-range of the normal
temperature region, where one would expect the greatest
metabolic efficiency, and greatest capacity to overcome an other
environmental hurdle by homeostatic mechanisms.

Box 1 A hypothetical physiological basis for the effect of temperature on microbial growth rate

Effect of Temperature on Enzyme Structure and Efficiency

The rate of enzyme-catalyzed reactions is also dependent on the concentration of active enzyme – itself a function of temperature. Enzymes are proteins. The
functional activity of enzymes is dependent upon their shape, or conformation, but they are flexible – the flexibility being required to achieve their catalytic function.
Temperature affects the bonds in the molecule and, if the temperature changes too much, the conformation becomes so distorted that the enzyme is no longer
catalytically active. This process is called ‘denaturation.’ Denaturation can be visualized as unfolding of the protein and can occur both when temperature becomes too
high and also when it becomes too low. That denaturation is reversible and the protein can refold spontaneously if the temperature returns to within the range for
stability. If the temperature becomes too high, however, irreversible denaturation takes place.
Hypothetical Physiological Basis of Temperature on Microbial Metabolism
A number of theoretical models have been advanced since the 1930s to explain the effect of temperature on bacterial growth rate. Most have as their basis the idea of
a rate limiting, enzyme-catalyzed, ‘master reaction’ for growth. The concept of the models for the temperature dependence of poikilothermic growth mentioned earlier
is that there is a single enzyme-catalyzed reaction that limits microbial growth rate under all conditions. This putative reaction and the enzyme that catalyzes it have
been termed the ‘master reaction’ and the ‘master enzyme,’ respectively. The activation energy of the master reaction is considered to be the ultimate limit to growth
rate at all temperatures.
The hypothesis continues that the master enzyme is subject to the effects of temperature, so that as temperature increases above the optimum for conformational
stability or decreases below it, the enzyme progressively becomes denatured. The transition of the master enzyme between conformationally active and inactive states
is a function of temperature. The effect of this is a reduced level of sites available for catalysis, perceived as a reduction in the rate of reaction as seen at high and low
temperature beyond the normal physiological range.
 ECOLOGY OF BACTERIA AND FUNGI IN FOODS j Influence of Temperature
If the lag time is a period of metabolic adjustment, requiring
synthesis of new protein, it follows that the effect of tempera-
ture on those processes will be similar to the effect of temper-
ature on growth rate. The induction of lag times due to abrupt
temperature shift corresponds to whether the temperature shift
involves a transition from one temperature region to another,
particularly from the normal to low regions.
Effects of Temperature on Metabolism
Enzymes energetically stabilize transition states of reaction
intermediates. By catalyzing specific reactions between selected
substrates, enzymes can act as ‘molecular switches’ in deter-
mining both the rate and the direction of metabolic pathways.
The preceding discussion has described how temperature
may act at a fundamental level of metabolism by directly
influencing the type and rate of biochemical reactions. Tem-
perature is an ‘extrinsic’ ecological factor influencing microbial
growth and metabolism. Although most bacteria and fungi
can actively regulate ‘intrinsic’ physicochemical parameters
within the cell (e.g., water activity, pH, redox), the microbial
cell can react only to changes in environmental temperature in
an effort to maintain functional metabolism. The regulation of
cellular metabolism in response to changes in environmental
temperature is of primary importance to survival and growth.
There are two broad areas in which metabolic regulation in
response to temperature variation occurs:
l The microbial cell, as a single compartment, must maintain
functional integrity and continue to provide a suitable
physicochemical environment for metabolic function.
l The regulation of enzyme activity must maintain coordi-
nation between catabolic and anabolic processes.
Maintenance of a physicochemical environment compat-
ible with enzyme function necessitates a functional cell
membrane assembly for both bacteria and fungi. Under the
fluid mosaic membrane model, this requires the following:
l That the cell regulates its lipid composition to ensure that
a stable bilayer is formed with membrane proteins, and
l That this bilayer remains in a sufficiently ‘fluid’ state.
A great deal of research has been aimed at the latter
requirement. The former point has been ignored largely due to
the general assumption that natural lipid mixtures spontane-
ously form a bilayer arrangement. Generally, during normal
cell growth, this is true.
Lipid Composition
In the fluid state, the lipid components of a membrane bilayer
remain miscible in all proportions. However, as the fluid-
crystalline phase transition occurs during a temperature
downshift (Figure 7) individual lipid components begin to
separate and crystallize depending on their individual ther-
modynamic properties. As the bilayer freezes, crystalline
regions grow at the expense of fluid ones, with progressively
lower melting point lipids moving out of fluid regions into
crystalline ones.
The crystallization, or freezing, of the membrane causes
large changes in viscoelastic properties of the cell. In the
607
(a)
Tm
Th
(c)
(b)
Figure 7 Illustration of the lipid bilayer structure in (a) the crystalline
(gel) phase where acyl motion is low; (b) the fluid phase where acyl
residue motion is high; and (c) a nonbilayer (inverted hexagonal)
lipid phase. Tm refers to the liquid–crystalline phase transition that may
occur to the membrane due to a temperature downshift. Th refers to
the bilayer–nonbilayer phase transition that may occur due to
a temperature upshift.
predominantly crystalline state, cellular membranes can become
‘osmotically fragile’ with leakage of intracellular components as
the bilayer loses its ability to act as an efficient semipermeable
barrier. Such leakage is believed to occur from two main sources:
the formation of microscopic fissures in crystalline regions and
the formation of grain boundary effects. Grain boundary effects
represent areas of disorder occurring at interfaces between
differently oriented crystalline regions formed during the fluid-
crystalline phase transition. These areas may provide leakage
sites for small molecules and ions. In addition, the crystalline
regions formed are, in effect, semisolid regions within the
membrane (Figure 7). As the membrane loses the flexibility
previously afforded by the more viscous fluid state, mechanical
deformation, and shrinkage can result in microscopic fissures
forming through which additional leakage may occur.
Second, the physical state of the membrane lipids also
has the potential to exert a large effect on many essential
physiological cell processes, such as sugar, amino acid, and
ion transport, chemotaxis, and membrane-associated oxida-
tion and reduction enzymes. The sensitivity of these pro-
cesses to the physical state of the membrane derives from the
fact that major protein components or assemblies of these
systems are located within, or in close association with, the
lipid bilayer.
Liquid–crystalline phase transitions caused by temperature
downshifts therefore are detrimental for the cell. Indeed, for
almost all microbes, the membrane bilayer is present in a fluid
or predominantly fluid state at growth temperature. For
example, in E. coli, the bulk crystalline–liquid phase transition is
completed 7–15
C below the ambient temperature, ensuring
that the membrane is completely fluid at the temperature of
608 ECOLOGY OF BACTERIA AND FUNGI IN FOODS j Influence of Temperature
growth. Upshifts in temperature may also be detrimental to
metabolic function, as the membrane may lose proper function
from excess fluidity. If the temperature upshift is of sufficient
severity, the increased fatty acid chain motion within the bilayer
may also result in the formation of nonbilayer lipid phases and
the loss of membrane function (Figure 7). The transition
temperature is controlled mainly by the melting point of the
constituent membrane phospholipid fatty acids, as the phase
transition is essentially a hydrocarbon-mediated event. By
manipulation of phospholipid fatty acid composition, bacteria
and fungi may alter their physical membrane characteristics to
maintain metabolic functions during and after changes in
environmental temperature.
Regulation of Enzymes in Response to Temperature
The manipulation by bacteria and fungi of one aspect of their
cellular composition, to retain functionality in response to
changes in temperature, has been described in general terms.
The innate effect of temperature on enzyme function and the
critical role of enzymes in metabolic processes has also been
discussed. There are multiple mechanisms by which metabolic
regulation occurs. Enzymes also play a fundamental role in the
regulation of metabolism in response to temperature. Two
major modes of enzyme regulation occur:
l A change in the cellular concentration of a given enzyme
(usually achieved by a change in the rate of enzyme
synthesis), thereby altering the overall level of cellular
activity, and
l Modulation of existing enzyme activity primarily via cova-
lent modification or allosteric interactions.
An example of such an enzymatic regulatory mechanism is
found in the adaptation of membrane fatty acid composition
Acetyl-ACP
+ Malonyl-ACP
3 Dehydration
via HDD 1 Condensation via
KAS I or KAS III
2 Reduction
via KAS R
, Dehydration 4 Reduction and elongation
product by KAS I or KAS III
, Dehydration 4 Elongation by
product KAS I
Figure 8 Schematic pathway of fatty acid biosynthesis in E. coli denoting the major products involved in thermal regulation. The synthesis of cis-vaccenic
acid is a critical metabolic function in response to decreasing environmental temperature. This process is regulated by the activity of KAS II. Abbreviations:
KAS I, b-ketoacyl-ACP synthase I; KAS II, b-ketoacyl-ACP synthase II; KAS III, b-ketoacyl-ACP synthase III; KAS R, b-ketoacyl-ACP reductase; HDD,
hydroxydecanoyl-ACP dehydrase.
to temperature within E. coli. Figure 8 summarizes the major
steps in the fatty acid biosynthetic pathway of E. coli.
The major fatty acids produced by E. coli are palmitic acid
(16:0), palmitoleic acid (16:1u7c), or cis-vaccenic acid
(18:1u7c) with the ratio of these major components varying
with temperature. In response to a decrease in growth
temperature, the amount of cis-vaccenic acid in the membrane
increases while the level of palmitic acid declines. That is, there
is an increase in unsaturated fatty acids at the expense of
saturated components.
Studies with temperature sensitive mutants of E. coli have
revealed that the specific ability to produce cis-vaccenic acid
(rather than simply palmitoleic acid) was essential for thermal
regulation. Further investigations revealed the presence of
a single enzyme, b-ketoacyl-ACP synthase II (KAS II) was
responsible for the critical step of converting palmitoleic acid to
cis-vaccenic acid during low-temperature thermal regulation
(Figure 8). Inhibitor studies demonstrated the neither mRNA
nor protein synthesis was required to achieve an increased rate
of cis-vaccenic acid synthesis within 30 s of a temperature
downshift. That is, KAS II is present within E. coli at all growth
temperatures but is regulated so that it becomes active only
under low-temperature conditions.
Similar studies of fatty acid modification in fungi have
highlighted a further important aspect of metabolic response to
temperature. Experiments using the mycelial fungi Tetrahymena
pyriformis and Cunninghamella japonica indicate that the
production of unsaturated fatty acids by enzyme-linked desa-
turation relies on the temperature-induced changes in
membrane fluidity. A decrease in membrane fluidity (with
decreasing temperature) results in the activation of membrane-
associated desaturase enzymes, which undergo a change in
conformation allowing them to act on surrounding fatty acid
substrates. The increase in unsaturated fatty acids consequently
Palmitic acid
(16:0)
Palmitoleic acid cis-Vaccenic acid
(16:1 7 c) (18:1 7 c)
5 Elongation by
KAS II
 ECOLOGY OF BACTERIA AND FUNGI IN FOODS j Influence of Temperature
restores a functional level of fluidity to the membrane.
Importantly, in this case, the activation of desaturase enzymes
is modulated directly by the degree of membrane fluidity,
which is influenced by temperature.
Microbes display both generic and specific responses to
extremes of temperature. Suites of proteins (often involved in
protection of enyzme conformation) are produced in response
to temperature changes and described as heat-shock, or cold-
shock, proteins. Specific responses to temperature change also
are documented. In the intracellular pathogen L. monocytogenes,
for example, virulence genes are expressed at 37
C, a temper-
ature that might signal to the organism that it is in a mamma-
lian host, but not at 25
C – a temperature more likely to be
associated with the organism being in the environment, and
needing to adopt a saprophytic lifestyle rather than expending
energy on the production of virulence factors involved in
intracellular survival and movement when they are not needed.
Complex systems of control of gene expression in response to
temperature exist and mechanisms by which temperature is
‘sensed’ also are being elucidated, including transmembrane
proteins involving histidine kinases, and which respond to the
thickness of the lipid bilayer. Temperature sensitive sequences
of RNA in mRNA molecules, termed ‘RNA thermometers,’ fold
into a conformation at low temperature that prevents their
access to the ribosome at low temperature, thereby preventing
expression of that gene.
Summary
Bacteria and fungi are unable to achieve temperature homeo-
stasis and, within a narrow range of temperature, their meta-
bolic rates respond to temperature in the same manner as
simple chemical reactions, increasing in rate with increasing
temperature. Beyond this range, the effects of temperature
become more pronounced requiring microorganisms to
manipulate their composition to minimize the effects of
temperature on their metabolism. Different species have
adapted to growth in different temperature ranges, but even
with the capacity for manipulation, most bacteria can grow
within a range of temperature that spans 35–40
C only and
fungi 25–30
C only. Those temperature tolerance limits may
be further reduced by other environmental constraints.
Although there is a complex interaction of growth rates and
tolerance ranges of microbes to temperature and other factors,
609
the known patterns of microbial response to temperature
enable the microbial ecology of foods to be reasonably well
understood and to enable that ecology to be manipulated by
temperature control.
See also: Clostridium: Clostridium perfringens; Escherichia coli:
Escherichia coli; Predictive Microbiology and Food Safety;
Ecology of Bacteria and Fungi: Influence of Available Water;
Ecology of Bacteria and Fungi in Foods: Effects of pH; Hurdle
Technology; Lipid Metabolism; Food Packaging with
Antimicrobial Properties; Freezing of Foods: Damage to
Microbial Cells; Freezing of Foods: Growth and Survival of
Microorganisms; Heat Treatment of Foods: Principles of
Canning; Heat Treatment of Foods: Spoilage Problems
Associated with Canning; Heat Treatment of Foods:
Ultra-High-Temperature Treatments; Heat Treatment of
Foods – Principles of Pasteurization; Heat Treatment of Foods:
Action of Microwaves; Heat Treatment of Foods: Synergy
Between Treatments; Microbiology of Sous-vide Products;
Preservatives: Traditional Preservatives – Organic Acids;
Thermal Processes: Pasteurization.
Further Reading
Berry, E.D., Foegeding, P.M., 1997. Cold adaptation and growth of microorganisms.
Journal of Food Protection 60, 1583–1594.
Biesta-Peters, E.G., Reij, M.W., Zwietering, M.H., Gorris, L.M., 2011. Comparing
nonsynergy gamma models and interaction models to predict growth of emetic
Bacillus cereus for combinations of pH and water activity values. Applied and
Environmental Microbiology 77, 5707–5715.
Ecosystems: microbes: food. Supplement to the Journal of Applied Bacteriology. In:
Board, R.G., Jones, D., Kroll, R.G., Pettipher, G.L. (Eds.), 1992. Society for Applied
Bacteriology Symposium Series, No. 21, vol. 73. (Entire Issue.). Blackwell Scientific
Publications, Oxford.
Christopherson, P.H., Hensel, H., 1973. Temperature and Life. Springer-Verlag, Berlin,
p. 779.
Mossel, D.A.A., Corry, J.E.L., Struijik, C.B., Baird, R.M., 1995. Essentials of the
Microbiology of Foods. A Textbook for Advanced Studies. John Wiley and Sons,
Chichester, p. 699.
Neidhart, F.C., Ingraham, J.L., Schaechter, M., 1990. Physiology of the Bacterial Cell.
A Molecular Approach. Sinauer Associates Inc, Sunderland, Massachusetts,
p. 506.
Ratkowsky, D.A., Olley, J., Ross, T., 2005. Unifying temperature effects on the growth
rate of bacteria and the stability of globular proteins. Journal of Theoretical Biology
233, 351–362.