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Nutrition and Growth of Bacteria
©1997 Kenneth Todar University of Wisconsin Department of Bacteriology
Edited on May 28, 2000 by KennethTodar University of Wisconsin-Madison Department of Bacteriology
http://www.bact.wisc.edu/Bact303/Controlofmicrobialgrowth

Every organism must find in its environment all of the substances required for energy generation and cellular biosynthesis. The chemicals and elements of this environment that are utilized for bacterial growth are referred to as nutrients or nutritional requirements. In the laboratory, bacteria are grown in culture media which are designed to provide all the essential nutrients in solution for bacterial growth.
At an elementary level, the nutritional requirements of a bacterium such as E. coli are revealed by the cell's elemental composition, which consists of C, H, O, N, S. P, K, Mg, Fe, Ca, Mn, and traces of Zn, Co, Cu, and Mo. These elements are found in the form of water, inorganic ions, small molecules, and macromolecules which serve either a structural or functional role in the cells. The general physiological functions of the elements are outlined in the Table below.
Table 1. Major elements, their sources and functions in bacterial cells.
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Element
% of dry weight
Source
Function

Carbon
50
organic compounds or CO2
Main constituent of cellular material

Oxygen
20
H2O, organic compounds, CO2, and O2
Constituent of cell material and cell water; O2 is electron acceptor in aerobic respiration

Nitrogen
14
NH3, NO3, organic compounds, N2
Constituent of amino acids, nucleic acids nucleotides, and coenzymes

Hydrogen
8
H2O, organic compounds, H2
Main constituent of organic compounds and cell water

Phosphorus
3
inorganic phosphates (PO4)
Constituent of nucleic acids, nucleotides, phospholipids, LPS, teichoic acids

Sulfur
1
SO4, H2S, So, organic sulfur compounds
Constituent of cysteine, methionine, glutathione, several coenzymes

Potassium
1
Potassium salts
Main cellular inorganic cation and cofactor for certain enzymes

Magnesium
0.5
Magnesium salts
Inorganic cellular cation, cofactor for certain enzymatic reactions

Calcium
0.5
Calcium salts
Inorganic cellular cation, cofactor for certain enzymes and a component of endospores

Iron
0.2
Iron salts
Component of cytochromes and certain nonheme iron-proteins and a cofactor for some enzymatic reactions


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The above table ignores the occurrence of trace elements in bacterial nutrition. Trace elements are metal ions required by certain cells in such small amounts that it is difficult to detect (measure) them, and it is not necessary to add them to culture media as nutrients. Trace elements are required in such small amounts that they are present as "contaminants" of the water or other media components. As metal ions, the trace elements usually act as cofactors for essential enzymatic reactions in the cell. One organism's trace element may be another's required element and vice-versa, but the usual cations that qualify as trace elements in bacterial nutrition are Mn, Co, Zn, Cu, and Mo.
In order to grow in nature or in the laboratory, a bacterium must have an energy source, a source of carbon and other required nutrients, and a permissive range of physical conditions such as O2 concentration, temperature, and pH. Sometimes bacteria are referred to as individuals or groups based on their patterns of growth under various chemical (nutritional) or physical conditions. For example, phototrophs are organisms that use light as an energy source; anaerobes are organisms that grow without oxygen; thermophiles are organisms that grow at high temperatures.
Carbon and Energy Sources for Bacterial Growth
All living organisms require a source of energy.
Organisms that use radiant energy (light) are called phototrophs.
Organisms that use (oxidize) an organic form of carbon are called heterotrophs or chemo(hetero)trophs.
Organisms that oxidize inorganic compounds are called lithotrophs.
The carbon requirements of organisms must be met by organic carbon (a chemical compound with a carbon-hydrogen bond) or by CO2. Organisms that use organic carbon are heterotrophs and organisms that use CO2 as a sole source of carbon for growth are called autotrophs.
Thus, on the basis of carbon and energy sources for growth four major nutritional types of procaryotes may be defined (Table 2)
Table 2. Major nutritional types of procaryotes.

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Nutritional Type
Energy Source
Carbon Source
Examples

Photoautotrophs
Light
CO2
Cyanobacteria, some Purple and Green Bacteria

Photoheterotrophs
Light
Organic compounds
Some Purple and Green Bacteria

Chemoautotrophs or Lithotrophs (Lithoautotrophs)
Inorganic compounds, e.g. H2, NH3, NO2, H2S
CO2
A few Bacteria and many Archaea

Chemoheterotrophs or Heterotrophs
Organic compounds
Organic compounds
Most Bacteria, some Archaea


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Almost all eukaryotes are either photoautotrophic (e.g. plants and algae) or heterotrophic (e.g. animals, protozoa, fungi). Lithotrophy is unique to procaryotes and photoheterotrophy, common in the purple and green Bacteria, occurs only in a very few eukaryotic algae. Phototrophy has not been found in the Archaea.
This simplified scheme for use of carbon, either organic carbon or CO2, ignores the possibility that an organism, whether it is an autotroph or a heterotroph, may require small amounts of certain organic compounds for growth because they are essential substances that the organism is unable to synthesize from available nutrients. Such compounds are called growth factors.
Growth factors are required in small amounts by cells because they fulfill specific roles in biosynthesis. The need for a growth factor results from either a blocked or missing metabolic pathway in the cells. Growth factors are organized into three categories.
1. purines and pyrimidines: required for synthesis of nucleic acids (DNA and RNA)
2. amino acids: required for the synthesis of proteins
3. vitamins: needed as coenzymes and functional groups of certain enzymes
Some bacteria (e.g E. coli) do not require any growth factors: they can synthesize all essential purines, pyrimidines, amino acids and vitamins, starting with their carbon source, as part of their own intermediary metabolism. Certain other bacteria (e.g. Lactobacillus) require purines, pyrimidines, vitamins and several amino acids in order to grow. These compounds must be added in advance to culture media that are used to grow these bacteria. The growth factors are not metabolized directly as sources of carbon or energy, rather they are assimilated by cells to fulfill their specific role in metabolism. Mutant strains of bacteria that require some growth factor not needed by the wild type (parent) strain are referred to as auxotrophs. Thus, a strain of E. coli that requires the amino acid tryptophan in order to grow would be called a tryptophan auxotroph and would be designated E. coli trp-.
Some vitamins that are frequently required by certain bacteria as growth factors are listed in Table 3. The function(s) of these vitamins in essential enzymatic reactions gives a clue why, if the cell cannot make the vitamin, it must be provided exogenously in order for growth to occur.
Table 3. Common vitamins required in the nutrition of certain procaryotes.

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Vitamin
Coenzyme form
Function

p-Aminobenzoic acid (PABA)
-
Precursor for the biosynthesis of folic acid

Folic acid
Tetrahydrofolate
Transfer of one-carbon units and required for synthesis of thymine, purine bases, serine, methionine and pantothenate

Biotin
Biotin
Biosynthetic reactions that require CO2 fixation

Lipoic acid
Lipoamide
Transfer of acyl groups in oxidation of keto acids

Mercaptoethane-sulfonic acid
Coenzyme M
CH4 production by methanogens

Nicotinic acid
NAD (nicotinamide adenine dinucleotide) and NADP
Electron carrier in dehydrogenation reactions

Pantothenic acid
Coenzyme A and the Acyl Carrier Protein (ACP)
Oxidation of keto acids and acyl group carriers in metabolism

Pyridoxine (B6)
Pyridoxal phosphate
Transamination, deamination, decarboxylation and racemation of amino acids

Riboflavin (B2)
FMN (flavin mononucleotide) and FAD (flavin adenine dinucleotide)
Oxidoreduction reactions

Thiamine (B1)
Thiamine pyrophosphate (TPP)
Decarboxylation of keto acids and transaminase reactions

Vitamin B12
Cobalamine coupled to adenine nucleoside
Transfer of methyl groups

Vitamin K
Quinones and napthoquinones
Electron transport processes


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Culture Media for the Growth of Bacteria
For any bacterium to be propagated for any purpose it is necessary to provide the appropriate biochemical and biophysical environment. The biochemical (nutritional) environment is made available as a culture medium, and depending upon the special needs of particular bacteria (as well as particular investigators) a large variety and types of culture media have been developed with different purposes and uses. Culture media are employed in the isolation and maintenance of pure cultures of bacteria and are also used for identification of bacteria according to their biochemical and physiological properties.
The manner in which bacteria are cultivated, and the purpose of culture media, vary widely. Liquid media are used for growth of pure batch cultures while solidified media are used widely for the isolation of pure cultures, for estimating viable bacterial populations, and a variety of other purposes. The usual gelling agent for solid or semisolid medium is agar, a hydrocolloid derived from red algae. Agar is used because of its unique physical properties (it melts at 100 degrees and remains liquid until cooled to 40 degrees, the temperature at which it gels) and because it cannot be metabolized by most bacteria. Hence as a medium component it is relatively inert; it simply holds (gels) nutrients that are in aquaeous solution.
Culture media may be classified into several categories depending on their composition or use. A chemically-defined (synthetic) medium (Table 4a and 4b) is one in which the exact chemical composition is known. A complex (undefined) medium (Table 5a and 5b) is one in which the exact chemical constitution of the medium is not known. Defined media are usually composed of pure biochemicals off the shelf; complex media usually contain complex materials of biological origin such as blood or milk or yeast extract or beef extract, the exact chemical composition of which is obviously undetermined. A defined medium is a minimal medium (Table4a) if it provides only the exact nutrients (including any growth factors) needed by the organism for growth. The use of defined minimal media requires the investigator to know the exact nutritional requirements of the organisms in question. Chemically-defined media are of value in studying the minimal nutritional requirements of microorganisms, for enrichment cultures, and for a wide variety of physiological studies. Complex media usually provide the full range of growth factors that may be required by an organism so they may be more handily used to cultivate unknown bacteria or bacteria whose nutritional requirement are complex (i.e., organisms that require a lot of growth factors).
Most pathogenic bacteria of animals, which have adapted themselves to growth in animal tissues, require complex media for their growth. Blood, serum and tissue extracts are frequently added to culture media for the cultivation of pathogens. Even so, for a few fastidious pathogens such as Treponema pallidum, the agent of syphilis, and Mycobacterium leprae, the cause of leprosy, artificial culture media and conditions have not been established. This fact thwarts the the ability to do basic research on these pathogens and the diseases that they cause.
Other concepts employed in the construction of culture media are the principles of selection and enrichment. A selective medium is one which has a component(s) added to it which will inhibit or prevent the growth of certain types or species of bacteria and/or promote the growth of desired species. One can also adjust the physical conditions of a culture medium, such as pH and temperature, to render it selective for organisms that are able to grow under these certain conditions.
A culture medium may also be a differential medium if allows the investigator to distinguish between different types of bacteria based on some observable trait in their pattern of growth on the medium. Thus a selective, differential medium for the isolation of Staphylococcus aureus, the most common bacterial pathogen of humans, contains a very high concentration of salt (which the staph will tolerate) that inhibits most other bacteria, mannitol as a source of fermentable sugar, and a pH indicator dye. From clinical specimens, only staph will grow. S. aureus is differentiated from S. epidermidis (a nonpathogenic component of the normal flora) on the basis of its ability to ferment mannitol. Mannitol-fermenting colonies (S. aureus)produce acid which reacts with the indicator dye forming a colored halo around the colonies; mannitol non-fermenters (S. epidermidis) use other non-fermentative substrates in the medium for growth and do not form a halo around their colonies.
An enrichment medium employs a slightly different twist. An enrichment medium (Table 5a and 5b) contains some component that permits the growth of specific types or species of bacteria, usually because they alone can utilize the component from their environment. However, an enrichment medium may have selective features. An enrichment medium for nonsymbiotic nitrogen-fixing bacteria omits a source of added nitrogen to the medium. The medium is inoculated with a potential source of these bacteria (e.g. a soil sample) and incubated in the atmosphere wherein the only source of nitrogen available is N2. A selective enrichment medium (Table 5b) for growth of the extreme halophile (Halococcus) contains nearly 25 percent salt [NaCl], which is required by the extreme halophile and which inhibits the growth of all other procaryotes.
Table 4a. Minimal medium for the growth of Bacillus megaterium. An example of a chemically-defined medium for growth of a heterotrophic bacterium.

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Component
Amount
Function of component

sucrose
10.0 g
C and energy source

K2HPO4
2.5 g
pH buffer; P and K source

KH2PO4
2.5 g
pH buffer; P and K source

(NH4)2HPO4
1.0 g
pH buffer; N and P source

MgSO4 7H2O
0.20 g
S and Mg++ source

FeSO4 7H2O
0.01 g
Fe++ source

MnSO4 H2O
0.007 g
Mn++ Source

water
985 ml


pH 7.0




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Table 4b. Defined medium (also an enrichment medium)for the growth of Thiobacillus thiooxidans, a lithoautotrophic bacterium.

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Component
Amount
Function of component

NH4Cl
0.52 g
N source

KH2PO4
0.28 g
P and K source

MgSO4 7H2O
0.25 g
S and Mg++ source

CaCl2 2H2O
0.07 g
Ca++ source

Elemental Sulfur
1.56 g
Energy source

C02
5%*
C source

water
1000 ml


pH 3.0




* Aerate medium intermittently with air containing 5% CO2.

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Table 5a. Complex medium for the growth of fastidious bacteria.

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Component
Amount
Function of component

Beef extract
1.5 g
Source of vitamins and other growth factors

Yeast extract
3.0 g
Source of vitamins and other growth factors

Peptone
6.0 g
Source of amino acids, N, S, and P

Glucose
1.0 g
C and energy source

Agar
15.0 g
Inert solidifying agent

water
1000 ml


pH 6.6




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Table 5b. Selective enrichment medium for growth of extreme halophiles.

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Component
Amount
Function of component

Casamino acids
7.5 g
Source of amino acids, N, S and P

Yeast extract
10.0 g
Source of growth factors

Trisodium citrate
3.0 g
C and energy source

KCl
2.0 g
K+ source

MgSO4 7 H2O
20.0 g
S and Mg++ source

FeCl2
0.023 g
Fe++ source

NaCl
250 g
Na+ source for halophiles and inhibitory to nonhalophiles

water
1000 ml


pH 7.4




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Physical and Environmental Requirements for Microbial Growth
The procaryotes exist in nature under an enormous range of physical conditions such as O2 concentration, Hydrogen ion concentration (pH) and temperature. The exclusion limits of life on the planet, with regard to environmental parameters, are always set by some microorganism, most often a procaryote, and frequently an Archaeon. Applied to all microorganisms is a vocabulary of terms used to describe their growth (ability to grow) within a range of physical conditions. A thermophile grows at high temperatures, an acidophile grows at low pH, an osmophile grows at high solute concentration, and so on. This nomenclature will be employed in this section to describe the response of the procaryotes to a variety of physical conditions.
The Effect of Oxygen
Oxygen is a universal component of cells and is always provided in large amounts by H2O. However, procaryotes display a wide range of responses to molecular oxygen O2 (Table 6).
Obligate aerobes require O2 for growth; they use O2 as a final electron acceptor in aerobic respiration.
Obligate anaerobes (occasionally called aerophobes) do not need or use O2 as a nutrient. In fact, O2 is a toxic substance, which either kills or inhibits their growth. Obligate anaerobic procaryotes may live by fermentation, anaerobic respiration, bacterial photosynthesis, or the novel process of methanogenesis.
Facultative anaerobes (or facultative aerobes) are organisms that can switch between aerobic and anaerobic types of metabolism. Under anaerobic conditions (no O2) they grow by fermentation or anaerobic respiration, but in the presence of O2 they switch to aerobic respiration.
Aerotolerant anaerobes are bacteria with an exclusively anaerobic (fermentative) type of metabolism but they are insensitive to the presence of O2. They live by fermentation alone whether or not O2 is present in their environment.
Table 6. Terms used to describe O2 Relations of Microorganisms.

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Environment


Group
Aerobic
Anaerobic
O2 Effect

Obligate Aerobe
Growth
No growth
Required (utilized for aerobic respiration)

Microaerophile
Growth if level not too high
No growth
Required but at levels below 0.2 atm

Obligate Anaerobe
No growth
Growth Toxic


Facultative Anaerobe (Facultative Aerobe)
Growth
Growth
Not required for growth but utilized when available

Aerotolerant Anaerobe
Growth
Growth
Not required and not utilized


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The response of an organism to O2 in its environment depends upon the occurrence and distribution of various enzymes which react with O2 and various oxygen radicals that are invariably generated by cells in the presence of O2. All cells contain enzymes capable of reacting with O2. For example, oxidations of flavoproteins by O2 invariably result in the formation of H202 (peroxide) as one major product and small quantities of an even more toxic free radical, superoxide or O2.-. Also, chlorophyll and other pigments in cells can react with O2 in the presence of light and generate singlet oxygen, another radical form of oxygen which is a potent oxidizing agent in biological systems.
In aerobes and aerotolerant anaerobes the potential for lethal accumulation of superoxide is prevented by the enzyme superoxide dismutase (Figure 1). All organisms which can live in the presence of O2 (whether or not they utilize it in their metabolism) contain superoxide dismutase. Nearly all organisms contain the enzyme catalase, which decomposes H2O2. Even though certain aerotolerant bacteria such as the lactic acid bacteria lack catalase, they decompose H2O2 by means of peroxidase enzymes which derive electrons from NADH2 to reduce peroxide to H2O. Obligate anaerobes lack superoxide dismutase and catalase and/or peroxidase, and therefore undergo lethal oxidations by various oxygen radicals when they are exposed to O2.
All photosynthetic (and some nonphotosynthetic) organisms are protected from lethal oxidations of singlet oxygen by their possession of carotenoid pigments which physically react with the singlet oxygen radical and lower it to its nontoxic "ground" (triplet) state. Carotenoids are said to "quench" singlet oxygen radicals.
Figure 1. The action of superoxide dismutase, catalase and peroxidase. These enzymes detoxify oxygen radicals that are inevitably generated by living systems in the presence of O2. The distribution of these enzymes in cells determines their ability to exist in the presence of O2.

Table 7. Distribution of superoxide dismutase, catalase and peroxidase in procaryotes with different O2 tolerances.

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Group
Superoxide dismutase
Catalase
Peroxidase

Obligate aerobes and most facultative anaerobes (e.g. Enterics)
+
+
-

Most aerotolerant anaerobes (e.g. Streptococci)
+
-
+

Obligate anaerobes (e.g. Clostridia, Methanogens, Bacteroides)

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The Effect of pH on Growth
The pH, or hydrogen ion concentration, [H+], of natural environments varies from about 0.5 in the most acidic soils to about 10.5 in the most alkaline lakes. Appreciating that pH is measured on a logarithmic scale, the [H+] of natural environments varies over a billion-fold and some microorganisms are living at the extremes, as well as every point between the extremes! Most free-living procaryotes can grow over a range of 3 pH units, about a thousand fold change in [H+]. The range of pH over which an organism grows is defined by three cardinal points: the minimum pH, below which the organism cannot grow, the maximum pH, above which the organism cannot grow, and the optimum pH, at which the organism grows best. For most bacteria there is an orderly increase in growth rate between the minimum and the optimum and a corresponding orderly decrease in growth rate between the optimum and the maximum pH, reflecting the general effect of changing [H+] on the rates of enzymatic reaction (Figure 2).
Microorganisms which grow at an optimum pH well below neutrality (7.0) are called acidophiles. Those which grow best at neutral pH are called neutrophiles and those that grow best under alkaline conditions are called alkaliphiles. Obligate acidophiles, such as some Thiobacillus species, actually require a low pH for growth since their membranes dissolve and the cells lyse at neutrality. Several genera of Archaea, including Sulfolobus and Thermoplasma, are obligate acidophiles. Among eukaryotes, many fungi are acidophiles, and the champion of growth at low pH is the eukaryotic alga Cyanidium which can grow at a pH of 0.
In the construction and use of culture media, one must always consider the optimum pH for growth of a desired organism and incorporate buffers in order to maintain the pH of the medium in the changing milieu of bacterial waste products that accumulate during growth. Many pathogenic bacteria exhibit a relatively narrow range of pH over which they will grow. Most diagnostic media for the growth and identification of human pathogens have a pH near 7.
Figure 2. Growth rate vs pH for three environmental classes of procaryotes. Most free-living bacteria grow over a pH range of about three units. Note the symmetry of the curves below and above the optimum pH for growth.

Table 8. Minimum, maximum and optimum pH for growth of certain procaryotes.

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Organism
Minimum pH
Optimum pH
Maximum pH

Thiobacillus thiooxidans
0.5
2.0-2.8
4.0-6.0

Sulfolobus acidocaldarius
1.0
2.0-3.0
5.0

Bacillus acidocaldarius
2.0
4.0
6.0

Zymomonas lindneri
3.5
5.5-6.0
7.5

Lactobacillus acidophilus
4.0-4.6
5.8-6.6
6.8

Staphylococcus aureus
4.2
7.0-7.5
9.3

Escherichia coli
4.4
6.0-7.0
9.0

Clostridium sporogenes
5.0-5.8
6.0-7.6
8.5-9.0

Erwinia caratovora
5.6
7.1
9.3

Pseudomonas aeruginosa
5.6
6.6-7.0
8.0

Thiobacillus novellus
5.7
7.0
9.0

Streptococcus pneumoniae
6.5
7.8
8.3

Nitrobacter spp
6.6
7.6-8.6
10.0


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The Effect of Temperature on Growth
Microorganisms have been found growing in virtually all environments where there is liquid water, regardless of its temperature. In 1966, Professor Thomas D. Brock at Indiana University, made the amazing discovery in boiling hot springs of Yellowstone National Park that bacteria were not just surviving there, they were growing and flourishing. Boiling temperature could not inactivate any essential enzyme. Subsequently, procaryotes have been detected growing around black smokers and hydrothermal vents in the deep sea at temperatures at least as high as 115 degrees. Microorganisms have been found growing at very low temperatures as well. In supercooled solutions of H2O as low as -20 degrees, certain organisms can extract water for growth, and many forms of life flourish in the icy waters of the Antarctic, as well as household refrigerators, near 0 degrees.
A particular microorganism will exhibit a range of temperature over which it can grow, defined by three cardinal points in the same manner as pH (Figure 3, cf. Figure 2). Considering the total span of temperature where liquid water exists, the procaryotes may be subdivided into several subclasses on the basis of one or another of their cardinal points for growth. For example, organisms with an optimum temperature near 37 degrees (the body temperature of warm-blooded animals) are called mesophiles. Organisms with an optimum T between about 45 degrees and 70 degrees are thermophiles. Some Archaea with an optimum T of 80 degrees or higher and a maximum T as high as 115 degrees, are now referred to as extreme thermophiles or hyperthermophiles. The cold-loving organisms are psychrophiles defined by their ability to grow at 0 degrees. A variant of a psychrophile (which usually has an optimum T of 10-15 degrees) is a psychrotroph, which grows at 0 degrees but displays an optimum T in the mesophile range, nearer room temperature. Psychrotrophs are the scourge of food storage in refrigerators since they are invariably brought in from their mesophilic habitats and continue to grow in the refrigerated environment where they spoil the food. Of course, they grow slower at 2 degrees than at 25 degrees. Think how fast milk spoils on the counter top versus in the refrigerator.
Psychrophilic bacteria are adapted to their cool environment by having largely unsaturated fatty acids in their plasma membranes. Some psychrophiles, particularly those from the Antarctic have been found to contain polyunsaturated fatty acids, which generally do not occur in procaryotes. The degree of unsaturation of a fatty acid correlates with its solidification T or thermal transition stage (i.e., the temperature at which the lipid melts or solidifies); unsaturated fatty acids remain liquid at low T but are also denatured at moderate T; saturated fatty acids, as in the membranes of thermophilic bacteria, are stable at high temperatures, but they also solidify at relatively high T. Thus, saturated fatty acids (like butter) are solid at room temperature while unsaturated fatty acids (like canola oil) remain liquid in the refrigerator. Whether fatty acids in a membrane are in a liquid or a solid phase affects the fluidity of the membrane, which directly affects its ability to function. Psychrophiles also have enzymes that continue to function, albeit at a reduced rate, at temperatures at or near 0 degrees. Usually, psychrophile proteins and/or membranes, which adapt them to low temperatures, do not function at the body temperatures of warm-blooded animals (37 degrees) so that they are unable to grow at even moderate temperatures.
Thermophiles are adapted to temperatures above 60 degrees in a variety of ways. Often thermophiles have a high G + C content in their DNA such that the melting point of the DNA (the temperature at which the strands of the double helix separate) is at least as high as the organism's maximum T for growth. But this is not always the case, and the correlation is far from perfect, so thermophile DNA must be stabilized in these cells by other means. The membrane fatty acids of thermophilic bacteria are highly saturated allowing their membranes to remain stable and functional at high temperatures. The membranes of hyperthermophiles, virtually all of which are Archaea, are not composed of fatty acids but of repeating subunits of the C5 compound, phytane, a branched, saturated, "isoprenoid" substance, which contributes heavily to the ability of these bacteria to live in superheated environments. The structural proteins (e.g. ribosomal proteins, transport proteins (permeases) and enzymes of thermophiles and hyperthermophiles are very heat stable compared with their mesophilic counterparts. The proteins are modified in a number of ways including dehydration and through slight changes in their primary structure, which accounts for their thermal stability.
Figure 3. Growth rate vs temperature for four environmental classes of bacteria. Most bacteria will grow over a temperature range of about 30 degrees. The curves exhibit three cardinal points: minimum, optimum and maximum temperatures for growth. There is a steady increase in growth rate between the minimum and optimum temperatures, but slightly past the optimum a critical thermolabile cellular event occurs, and the growth rates plunge rapidly as the maximum T is approached.


Table 9. Terms used to describe microorganisms in relation to temperature requirements for growth.

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Temperature for growth (degrees C)
Group
Minimum
Optimum
Maximum
Comments

Psychrophile
Below 0
10-15
Below 20
Grow best at relatively low T

Psychrotroph
0
15-30
Above 25
Able to grow at low T but prefer moderate T

Mesophile
10-15
30-40
Below 45
Most bacteria esp. those living in association with warm-blooded animals

Thermophile
45
50-85
Above 100 (boiling)
Among all thermophiles is wide variation in optimum and maximum T


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Table 10a. Minimum, maximum and optimum temperature for growth of bacteria.

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Temperature for growth (degrees C)
Bacterium
Minimum
Optimum
Maximum

Listeria monocytogenes
1
30-37
45

Vibrio marinus
4
15
30

Pseudomonas maltophilia
4
35
41

Thiobacillus novellus
5
25-30
42

Staphylococcus aureus
10
30-37
45

Escherichia coli
10
37
45

Clostridium kluyveri
19
35
37

Streptococcus pyogenes
20
37
40

Streptococcus pneumoniae
25
37
42

Bacillus flavothermus
30
60
72

Thermus aquaticus
40
70-72
79

Methanococcus jannaschii
60
85
90

Sulfolobus acidocaldarius
70
75-85
90

Pyrobacterium brockii
80
102-105
115


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Table 10b. Optimum growth temperature of some procaryotes.

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Genus and species
Optimal growth temp (degrees C)

Vibrio cholerae
18-37

Photobacterium phosphoreum
20

Rhizobium leguminosarum
20

Streptomyces griseus
25

Rhodobacter sphaeroides
25-30

Pseudomonas fluorescens
25-30

Erwinia amylovora
27-30

Staphylococcus aureus
30-37

Escherichia coli
37

Mycobacterium tuberculosis
37

Pseudomonas aeruginosa
37

Streptococcus pyogenes
37

Treponema pallidum
37

Thermoplasma acidophilum
59

Thermus aquaticus
70

Bacillus caldolyticus
72

Pyrococcus furiosus
100


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Table 10c. Hyperthermophilic Archaea.

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Temperature for growth(degrees C)


Genus
Minimum
Optimum
Maximum
Optimum pH

Sulfolobus
55
75-85
87
2-3

Desulfurococcus
60
85
93
6

Methanothermus
60
83
88
6-7

Pyrodictium
82
105
113
6

Methanopyrus
85
100
110
7


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Water Availability
Water is the solvent in which the molecules of life are dissolved, and the availability of water is therefore a critical factor that affects the growth of all cells. The availability of water for a cell depends upon its presence in the atmosphere (relative humidity) or its presence in solution or a substance (water activity). The water activity (Aw) of pure H2O is 1.0 (100% water). Water activity is affected by the presence of solutes such as salts or sugars, that are dissolved in the water. The higher the solute concentration of a substance, the lower is the water activity and vice-versa. Microorganisms live over a range of Aw from 1.0 to 0.7. The Aw of human blood is 0.99; seawater = 0.98; maple syrup = 0.90; Great Salt Lake = 0.75. Water activities in agricultural soils range between 0.9 and 1.0.
The only common solute in nature that occurs over a wide concentration range is salt [NaCl], and microorganisms are named based on their growth response to salt. Microorganisms that require some NaCl for growth are halophiles. Mild halophiles require 1-6% salt, moderate halophiles require 6-15% salt; extreme halophiles require 15-30% NaCl for growth. Bacteria that are able to grow at moderate salt concentrations, even though they grow best in the absence of NaCl, are called halotolerant. Although halophiles are "osmophiles" (and halotolerant organisms are "osmotolerant") the term osmophiles is usually reserved for organisms that are able to live in environments high in sugar. Organisms which live in dry environments (made dry by lack of water) are called xerophiles.
The concept of lowering water activity in order to prevent bacterial growth is the basis for preservation of foods by drying (in sunlight or by evaporation) or by addition of high concentrations of salt or sugar.
Figure 4. Growth rate vs osmolarity for different classes of procaryotes. Osmolarity is determined by solute concentration in the environment. Osmolarity is inversely related to water activity (Aw), which is more like a measure of the concentration of water (H2O) in a solution. Increased solute concentration means increased osmolarity and decreased Aw. a is the growth rate of a normal (nonhalophile) such as E. coli or Pseudomonas. b is the growth rate of a halotolerant bacterium such as Staphylococcus aureus. c is the growth rate of an extreme halophile such as Halobacterium, the predominant organism in the Great Salt Lake.


Table 11. Limiting water activities (Aw) for growth of certain procaryotes.

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Organism
Minimum Aw for growth

Caulobacter
1.00

Spirillum
1.00

Pseudomonas
.91

Salmonella/E. coli
.91

Lactobacillus
.90

Bacillus
.90

Staphylococcus
.85

Halobacterium
.75


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