Bacterial Nutrition, Growth and Metabolism Dr. Halide L.

Abella

Energy Metabolism

Overview

Metabolism
The sum of all chemical processes carried out by living organisms It includes: anabolism and catabolism

Anabolism
Reactions that require energy to synthesize complex molecules from simpler ones Needed for growth, reproduction and repair of cellular structures.

Overview

Catabolism
Reactions that release energy by breaking complex molecules into simpler one Provides an organism with energy for its life processes, including movement, transport and the synthesis of complex molecules (anabolism) All catabolic reactions involve electron transfer Allows energy to be captured in high-energy bonds in ATP and other similar molecules.

Overview

Oxidation
Loss or removal of electrons When a substance losses electrons, it is oxidized and energy is released. Many substances combine with oxygen x Transfer their electrons to oxygen x Oxygen need not be present if another electron acceptor is available

Reduction
Gain of electrons When a substance gains electrons, it is reduced Oxidation and reduction must occur simultaneously. The reactions are sometimes called redox reactions.

Energy and Work

Energy

The capacity to do work or to cause particular changes. All physical and chemical processes are the result of the application or movement of energy.

Three major types of work carried out by living cells

Chemical work Transport work

Chemical Work

y y

Mechanical work Involves the synthesis of complex biological molecules required by cells from much simpler precursors. Energy is needed to increase the molecular complexity of a cell.

Transport Work

Transport of molecules and ions across cell membranes against an electrochemical gradient Requires energy input Function:

Take up nutrients Eliminate wastes Maintain ion balances

Mechanical Work

y y

Occurs when there changes in the physical location of organisms, cells, and structures within cells. Requires energy input.

ATP

Adenosine triphosphate Energy currency of the cell

ATP

When ATP breaks down to ADP, energy is made available for useful work Energy from photosynthesis, aerobic respiration, anaerobic respiration, and fermentation is used to resynthesize ATP from ADP and Pi. An energy cycle is created in the cell

Classificiation of All Microorganisms According To Energy and Carbon Source

Chemo-heterotrophs Photo-heterotrophs Heterotrophs/Organotrophs Chemo-autotrophs Photo-autotrophs Autotrophs/Lithotrophs

All microorganisms

Autotrophic/Lithotrophic Bacteria

Utilize carbon dioxide as sole source of carbon Synthesize from the CO2 all the carbon skeletons of all their organic metabolites Require only water, inorganic salts, and CO2 for growth. Energy is derived either from light or from oxidation of one or more inorganic substances Unable to utilize CO2 as the sole source of carbon Require carbon in an organic form such as glucose Energy is derived either from light or from a portion of the organic compound All of the bacteria that cause disease in humans.

Heterotrophic/Organotrophic Bacteria
y y y y

Type Photolithotrophs

Carbon Source CO2

Energy Source Light

Electron Donor Inorganic compounds (H2S, S)

Examples =Photosynthetic bacteria -green sullfur -purple sulfur -cyanobacteria =Algae

Photoorganotrophs

Organic compounds

Light

Organic compounds

=Purple nonsulfur and =Green nonsulfur bacteria

Chemolithotrophs

CO2

Oxidationreduction reactions of inorganic compounds Oxidationreduction

Inorganic compounds (H2, S, H2S, Fe, NH3)

=Iron, Sulfur, Hydrogen and Nitrifying bacteria =Some Archaeobacteria =Pathogenic bacteria

Chemoorganotrophs

Organic compounds

Organic compounds

Metabolic Processes Among Heterotrophic Organisms

Scheme Aerobic Respiration Pathways Involved Glycolysis, TCA cycle, electron transport Final Electron Acceptor O2 Products ATP, CO2, H2O Chief Microbe Type Aerobes Facultative anaerobes

Anaerobic Metabolism Fermentation Glycolysis Organic molecules Various inorganic salts(NO3-, SO4-2, CO3-2) ATP, CO2, ethanol, lactic acid CO2, ATP, organic acids,H2S, CH4, N2 Facultative, aerotolerant, strict anaerobes Anaerobes; some facultatives

Anaerobic Respiration

Glycolysis, TCA cycle,electron transport

Aerobic Respiration

Glycolysis

Also called the Embden-Meyerhof-Parnas (EMP) pathway It does not require oxygen but can occur in either the presence of absence of oxygen Enzymatically converts glucose through several steps into pyruvic acid

A central metabolite Occupies an important position in several pathways

Alternatives to Glycolysis

Other metabolic pathways utilized by microorganisms for glucose oxidation Pentose Phosphate Pathway Entner-Doudoroff Pathway

Pentose Phosphate Pathway

y y y y

Phosphogluconate Pathway Utilized by Brucella abortus, species of Acetobacter, Escherichia coli and Bacillus subtilis Can function at the same time as glycolysis It breaks down not only glucose but also five- carbon sugars (pentoses).

Entner-Dourodoff Pathway

Carried out by Pseudomonas, Azobacter and Neisseria Replaces the glycolytic and pentose phosphate pathways

Fate of Pyruvic Acid (Pyruvate)

Fermentation

Incomplete oxidation of glucose or other carbohydrates in the absence of oxygen Uses organic compounds as the terminal electron acceptors Yields a small amount of ATP. Defined by bacteriologists as the formation of acid, gas, and other products by the action of various bacteria on pyruvic acid.

Fermentation and Biochemical Testing

Knowledge of fermentation products

Important in industrial production Important in identifying bacteria by biochemical tests

Specimens are grown in media containing various carbohydrates, and the production of acid or acid and gas is noted. Examples

Escherichia ferments the milk sugar, lactose Shigella and Proteus do not. Escherichia can be further differentiated from Shigella, which ferments glucose

Anaerobic Respiration

Anaerobic respiratory system

Functions like the aerobic cytochrome system Except it utilizes oxygen-containing salts, rather than free oxygen, as the final electron acceptor.

Nitrate (NO3-) and nitrite (NO2-) reduction systems

Seen in E. coli and species of Bacillus and Pseudomonas

Energy-Yielding Autotrophic Metabolism

In photoautotrophs

Photosynthesis

Photosynthesis
y y y

In chemoautotrophs

The capture of energy from light and the use of this energy to manufacture carbohydrates from carbon dioxide Photosynthesis occurs in green and purple bacteria, in cyanobacteria, in algae, and in higher plants. occurs in two parts the photo part, or the light reactions x light energy is converted to chemical energy the synthesis part, or the dark reactions x chemical energy is used to make organic molecules

Chemoautotrophs

Unable to carry out photosynthesis but can oxidize inorganic substances for energy

Control of Energy Metabolism

Genetic Regulation Metabolic Regulation Modulation of the Glycolytic Pathway

Genetic Regulation

Induction Catabolite Repression

1.

Induction

The synthesis of enzymes catalyzing a particular sequence is turned on or off, depending on the demands for that specific sequence.

Genetic Regulation
2. Catabolite repression

Sometimes called glucose effect Frequently observed when organisms are grown in glucose and some other rapidly metabolizable energy source. There is repression of synthesis of enzymes that would metabolize the added substrate less rapidly than glucose. Example

E. coli grown in a medium containing both glucose and lactose It uses glucose preferentially until the sugar is exhausted. Then after a short lag, growth resumes with lactose as the carbon source.

Metabolic Regulation

Adenylate Energy Charge

Adenine nucleotides (ATP, ADP & AMP) are strategically placed to regulate the entire metabolic economy of the cell Catabolic sequences contain regulatory enzymes that are either

Activated by ADP or AMP OR Inhibited by ATP

Modulation of the Glycolytic Pathway

Pasteur Effect

Pasteurization involves heating food to a temperature that kills diseasecausing microorganisms and substantially reduces the levels of spoilage organisms.

Less glucose is consumed The accumulation of lactate is decreased

Pasteur & The Wine-to-Vinegar Connection

Louis Pasteur was hired by French wine makers to uncover the causes of periodic spoilage in wines. Especially troublesome was the conversion of wine to vinegar and the resultant sour flavor. After extensively studying beer making and wine grapes, Pasteur concluded that wine, both fine and not-so-fine, was the result of microbial action on the juices of the grape and that wine disease was caused by contaminating organisms that produced undesirable products such as acid. Although he did not know it at the time, the bacterial contaminants responsible for the acidity of the spoiled wines were likely to be Acetobacter or Gluconobacter introduced by the grapes, air, or winemaking apparatus. Pasteur s far-reaching solution to the problem is still with us today mild heating, or pasteurization, of the grape juice to destroy the contaminants, followed by inoculation of the juice with a pure yeast culture.

Physiology of Bacterial Growth

Requirements for Growth

Uptake of Nutrients Bacterial Chemotaxis Growth of Bacterial Populations Bacterial Cell Cycle Differentiation in Bacterial Cells

Requirements For Growth

Essential Nutrients

Any substance, whether in elemental or molecular form, that must be provided to an organism 2 categories of essential nutrients

Macronutrients or macroelements Micronutrients or trace elements

Requirements for Growth

Macronutrients

Required by the microorganisms in relatively large amount

CHONSP are components of carbohydrates, lipids, proteins, and nucleic acids

The remaining macroelements exist in the cell as cations and play a variety of roles.

Potassium is required for activity by a number of enzymes, including some of those involved in protein synthesis. Calcium contributes to the heat resistance of bacterial endospores. Magnesium serves as a cofactor for many enzymes, complexes with ATP, and stabilizes ribosomes and cell membranes Iron (Fe2+ and Fe3+) is a part of cytochromes and a cofactor for enzymes and electron-carrying proteins.
Requirements for Growth

Micronutrients

Manganese, zinc, cobalt, molybdenum, nickel, and copper (trace elements) Needed by most cells Cells require such small amounts that contaminants in water, glassware, and regular media components often are adequate for growth. Normally a part of enzymes and cofactors, and they aid in the catalysis of reactions and maintenance of protein structure.

Zinc

present at the active site of some enzymes

Manganese

aids many enzymes that catalyze the transfer of phosphate groups.

Requirements for Growth

Molybdenum

required for nitrogen fixation

Carbon and Carbon Dioxide

The majority of carbon compounds involved in the normal structure and metabolism of all cells are organic. Heterotrophs Organisms that must obtain its carbon in an organic form. Organic carbon originates from the bodies of other organisms Heterotrophs are dependent on other life forms Among the common organic molecules that can satisfy their carbon requirement are proteins, carbohydrates, lipids, and nucleic acids. In most cases, these nutrients provide several other elements as well. Autotrophs Organisms that uses CO2, an inorganic gas, as its carbon source. Because autotrophs have the special capacity to convert CO2 into organic compounds, they are not nutritionally dependent on other living things.
Requirements for Growth

Carbon and Carbon Dioxide

Some organisms require a higher concentrations (10%) of carbon dioxide than what is normally present in the atmosphere (0.03%). Neisseria and Brucella

Capnophiles

Carbon dioxide loving organisms They thrive under conditions of low oxygen and high carbon dioxide concentration.

Nitrogen

The main reservoir of nitrogen is nitrogen gas (N2) It makes up about 79% of the earth s atmosphere. This element is needed in the structure of proteins, DNA, RNA, and ATP. the primary sources of nitrogen for heterotrophs to be useful, they must first be degraded into their basic building blocks (proteins into amino acids; nucleic acids into nucleotides).
Requirements for Growth

Growth Factors

Many heterotrophic bacteria are unable to grow unless supplied with one or more growth factors Usually provided in the culture medium in the form of yeast extract or whole blood It includes B-complex vitamins, amino acids, purines and pyrimidines

Prototrophic organisms

Organisms that do not require an exogenous source of a given growth factor Capable of synthesizing their own Require the addition of growth factor to culture media in order for growth to occur.

Auxotrophic organisms

Requirements for Growth

Inorganic Ions

Small amounts of inorganic ions are required by all bacteria Magnesium Functions to stabilize ribosomes, cell membranes and nucleic acids Required for the activity of many enzymes Potassium Required for the activity of many enzymes In gram (+) organisms, it s composition in the cell is influenced by the teichoic acid content of the cell wall

Requirements for Growth

Oxygen

Classification of bacteria based on their oxygen requirements Obligate anaerobes Grow only under conditions of high reducing intensity Oxygen is toxic Aerotolerant anaerobes Not killed by exposure to oxygen Facultative anaerobes Capable of growth under both aerobic and anaerobic conditions Obligate aerobes Require oxygen for growth Microaerophilic organisms Grow best at low oxygen tensions High oxygen tension is inhibitory
Requirements for Growth

Oxygen

Requirements for Growth

Physical Requirements
1. Temperature

There is an optimal temperature at which the organism grows most rapidly and a range of temperatures over which growth can occur. Cellular division is specially sensitive to the damaging effects of high temperatures. Classification

Psychrophilic

-5 to 30 C, optimum at 10 to 20 C

Mesophilic

10 to 45 C, optimum at 20 to 40 C Human pathogens Requirements for Growth

Thermophilic

Physical Requirements
Thermophilic sulfur bacteria can live and grow in the runoff waters from such geysers despite the near-boiling temperatures.

Requirements for Growth

Physical Requirements
The temperature range over which an organism grows is determined largely by the temperatures at which its enzymes function. Within this temperature range, three critical temperatures can be identified:

1.

Minimum growth temperature

the lowest temperature at which cells can divide.

2.

Maximum growth temperature

the highest temperature at which cells can divide.

3.

Optimum growth temperature

the temperature at which cells divide most rapidly (shortest generation time)
Requirements for Growth

Physical Requirements
2. Hydrogen Ion Concentration

pH of the culture can affect the growth rate pH 7.2 to pH 7.6 optimal pH for most pathogenic bacteria Classification according to their tolerance for acidity and alkalinity

Acidophiles -pH range of 6.5 to 7.0 Neutrophiles -pH range of 7.5 to 8.0 Alkalophiles -pH range of 8.5 to 9.0

Requirements for Growth

Physical Requirements

Acidophiles

Lactobacillus produces lactic acid but tolerates only mild acidity Acid drips from long, hanging colonies of bacteria which have the consistency of strings of mucus.

Requirements for Growth

Physical Requirements

Neutrophiles

Most of the bacteria that cause human disease are neutrophiles

Alkalophiles

Vibrio cholerae, the causative agent of the disease cholera, grows best at a pH of about 9.0. Alcaligenes faecalis, which sometimes infects humans already weakened by another disease, can create and tolerate alkaline conditions of pH 9.0 or higher.

Many bacteria often produce sufficient quantities of acids as metabolic by-products that eventually interfere with their own growth. To prevent this situation in the laboratory cultivation of bacteria, buffers are incorporated into growth media to maintain the proper pH levels.
Requirements for Growth

Physical Requirements
3. Osmotic Conditions

The concentration of osmotically active solutes inside a bacterial cell is higher than the concentration outside the cell. Cells in such hyperosmotic environments lose water and undergo shrinking of the cell. Cells in distilled water have a higher osmotic pressure than their environment and, therefore, gain water. Cells fill with water and become distended. Majority of bacteria are osmotically tolerant

Except for the Mycoplasmas and other cell wall-defective organisms.

Their cell membranes contain transport systems that regulate the movement of dissolved substances across the membrane.
Requirements for Growth

Physical Requirements

Halophiles

Salt-loving organisms Require moderate to large quantities of salt (sodium chloride). Their membrane transport systems actively transport sodium ions out of the cells and concentrate potassium ions inside them. Typically found in the ocean, where the salt concentration (3.5%) is optimum for their growth. Extreme halophiles require salt concentrations of 20% to 30%. They are found in exceptionally salty bodies of water, such as the Dead Sea, and sometimes even in brine vats, where they cause spoilage of pickles being made.
Requirements for Growth