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Chapter 7: Microbial Physiology and Genetics        1




Chapter 7
Microbial Physiology and Genetics
Primary Objectives of the Chapter
Chapter 7 introduces aspects of microbial physiology, such as enzymes, metabolism (catabolic
and anabolic reactions), oxidation–reduction reactions, biochemical pathways, aerobic
respiration, and fermentation. Microbial genetic topics discussed in Chapter 7 include mutations,
the various ways in which bacteria acquire new genetic information (lysogenic conversion,
transduction, transformation, conjugation), genetic engineering, and gene therapy. The
information in Chapter 7 is considered essential in an introductory microbiology course.

Terms Introduced in This Chapter
After reading Chapter 7, you should be familiar with the following terms. These terms are
defined in Chapter 7 and in the Glossary.

Adenosine triphosphate (ATP)
Ames test
Anabolic reactions
Anabolism
Autotroph
Beneficial mutation
Catabolic reactions
Catabolism
Chemoautotroph
Chemoheterotroph
Chemolithotroph
Chemoorganotroph
Chemosynthesis
Chemotroph
Competence
Competent bacteria
Dehydrogenation reactions
Ecology
Ecosystem
Electron transport chain
Endoenzyme
Episome
Essential nutrients
Exoenzyme
Fermentation
Gene therapy
Chapter 7: Microbial Physiology and Genetics         2




Genetics
Glycolysis
Harmful mutation
Heterotroph
Krebs cycle
Lethal mutation
Lysogenic bacterium
Lysogenic conversion
Lysogeny
Metabolic reactions
Metabolite
Microbial physiology
Mutagen
Mutant
Mutation
Oxidation
Oxidation–reduction reactions
Phenotype
Photoautotroph
Photoheterotroph
Phototroph
Prophage
R-factor
Reduction
Silent mutation
Transduction
Transformation




       Review of Key Points
      Microbial physiology is the study of the life processes of microorganisms.
      Scientists have learned a great deal about cells—including human cells—by studying the
       nutritional needs of bacteria, their metabolic pathways, and why they live, grow,
       multiply, or die under certain conditions.
      All living organisms require sources of energy and carbon so that they can produce the
       molecules necessary for life. In addition, organisms must be provided with certain
       materials (called essential nutrients) that they themselves are unable to synthesize, but are
       required for survival; these essential nutrients vary from species to species.
      The energy source for certain organisms (called phototrophs) is light and for other
       organisms (called chemotrophs) is organic or inorganic chemicals.
      Chemolithotrophs and chemoorganotrophs are subcategories of chemotrophs.
       Chemolithotrophs (or simply lithotrophs) use inorganic chemicals as an energy source,
Chapter 7: Microbial Physiology and Genetics         3




    whereas chemoorganotrophs (or simply organotrophs) use organic chemicals as an
    energy source.
   An organism’s carbon source may be CO2 (in which case the organism is called an
    autotroph) or other organic compounds (in which case the organism is called a
    heterotroph). Humans, animals, protozoa, and fungi are heterotrophs, as are most
    bacteria.
   Interrelationships among the different nutritional types are of prime importance in the
    functioning of the ecosystem. Phototrophs (plants, algae, and certain bacteria) are the
    producers of food and oxygen for the chemoheterotrophs (animals). Dead plants and
    animals are recycled by the chemoheterotrophic saprophytic decomposers (certain fungi
    and bacteria) into nutrients for phototrophs and chemotrophs.
   Metabolism refers to all of the chemical reactions (metabolic reactions) that occur within
    a living cell, including the production of energy and the synthesis of new molecules.
   Metabolic reactions include catabolic reactions and anabolic reactions. Catabolic
    reactions (also called degradative reactions) involve the breaking of chemical bonds and
    the release of energy. Anabolic reactions (also called biosynthetic reactions) require
    energy because they involve the formation of chemical bonds.
   Most metabolic reactions are regulated by enzymes.
   Enzymes are biologic molecules (proteins) that serve as catalysts to control the rate of
    metabolic reactions. The enzymes produced by any particular cell are governed by the
    genotype of that cell, and the presence or absence of any particular enzyme is part of the
    phenotype of that cell.
   The substance upon which an enzyme acts is known at that enzyme’s substrate.
   All the enzymes that a cell is capable of producing need not be present in the cell at a
    given moment in time. They are produced to meet the metabolic needs of the cell, as
    determined by the internal and external environments.
   Endoenzymes are enzmes that remain within the cell that produced them, whereas
    exoenzymes are enzymes that leave the cell to catalyze reactions outside of the cell.
   Apoenzymes are proteins that are unable to catalyze reactions on their own. To catalyze
    reactions, apoenzymes must first link up with a cofactor (either a mineral ion or a
    coenzyme).
   An enzyme operates at peak efficiency within a particular pH and temperature range and
    when an appropriate concentration of the substrate for that enzyme exists. If the
    environment is too acidic, basic, hot, or cold, or contains too much or too little substrate,
    the enzyme will not operate at peak efficiency and the reaction will not proceed at its
    maximum rate.
   Catabolic reactions involve the breaking of chemical bonds and the release of energy.
    Anabolic reactions involve the formation of bonds, which requires energy.
   Adenosine triphosphate (ATP) is the principal energy-storing or energy-carrying
    molecule in the cell. Should a cell require energy, one of the high-energy bonds in an
    ATP molecule can be broken, producing energy, an ADP molecule, and a free phosphate.
    The energy can then be used for growth, reproduction, active transport of substances
    across membranes, sporulation, movement, anabolic reactions, and other energy-
    requiring activities.
Chapter 7: Microbial Physiology and Genetics         4




   Nutrients should be thought of as energy sources, and chemical bonds should be thought
    of as stored energy.
   A common pathway by which bacteria catabolize glucose is aerobic respiration, which
    consists of three phases: glycolysis, the Krebs cycle, and the electron transport chain.
    Most of the energy that is produced by aerobic respiration is produced by the electron
    transport chain. The breakdown of one molecule of glucose by aerobic respiration yields
    either 36 ATP molecules (procaryotic cells) or 38 ATP molecules (eucaryotic cells).
   Aerobes and facultative anaerobes are able to produce more energy than anaerobes,
    because they can catabolize glucose molecules via aerobic pathways. Anaerobes must
    catabolize glucose by fermentation, a relatively inefficient method that yields only two
    ATP molecules from a molecule of glucose.
   Oxygen does not participate in fermentation reactions.
   Oxidation reactions involve the loss of an electron, whereas reduction reactions involve
    the gain of an electron.
   Phototrophic organisms (algae, plants, and photosynthetic bacteria) derive their energy
    from the sun by photosynthesis. Chemosynthetic organisms use a chemical source of
    energy and raw materials to synthesize metabolites and macromolecules for growth and
    function of the organisms.
   As with humans, animals, and plants, the genetics of microbes involves DNA, genes, the
    genetic code, chromosomes, DNA replication, transcription, and translation—all part of
    molecular genetics.
   An organism’s genotype (or genome) is its complete collection of genes, whereas an
    organism’s phenotype is all the physical traits, attributes, or characteristics of the
    organism.
   Genes direct all functions of the cell, providing it with its own particular traits and
    individuality. An organism’s phenotype is the manifestation of that organism’s genotype.
   Constitutive genes are expressed at all times, whereas inducible genes are expressed only
    when the products that they code for (gene products) are needed.
   The base sequence of any gene on a chromosome may be altered accidentally in many
    ways, resulting in a mutation. Mutations are expressed not only in the cell in which the
    mutation occurred, but in subsequent generations as well. The altered genetic code will
    result in an altered protein, which could affect any of a number of different phenotypic
    characteristics (e.g., changes in colony characteristics, cell shape, biochemical activities,
    nutritional needs, antigenic sites, virulence, pathogenicity, drug resistance). Mutant
    bacteria are used in genetic and medical research and the production of vaccines.
   Mutations may be beneficial, harmful, or of no consequence to the cell or organism
    containing the mutation. Those of no consequence are called silent or neutral mutations.
   Beneficial mutations are of benefit to an organism, whereas harmful mutations result in
    the production of structurally altered proteins (often, nonfunctional enzymes). Some
    harmful mutations are lethal to the organism.
   Physical or chemical agents that cause an increased mutation rate are called mutagens.
Chapter 7: Microbial Physiology and Genetics        5




       In addition to mutations, genetic changes in a bacterial cell may be the result of lysogenic
        conversion, transduction, transformation, or conjugation, all of which occur in nature as
        well as in the laboratory.
       Lysogenic conversion involves temperate bacteriophages. In lysogenic conversion,
        bacteria gain new genetic information in the form of viral genes.
       Transduction also involves bacteriophages. In lysogenic conversion, bacteria gain new
        genetic information in the form of bacterial genes.
       In transformation, a bacterial cell becomes genetically transformed following uptake of
        DNA fragments (―naked DNA‖) from the environment.
       Conjugation involves the transfer of genetic material (usually a plasmid) from a donor
        cell to a recipient cell through a hollow sex pilus.
       A plasmid that contains multiple genes for antibiotic resistance is called a resistance
        factor or R-factor.
       The field of genetic engineering involves the introduction of new genes into cells. When
        a cell receives a new gene, it can produce the gene product that is coded for by that gene.
        Genetically engineered bacteria are used to produce products such as insulin, interferon,
        human growth hormone, and materials for use as vaccines.
       Gene therapy involves the use of viruses and plasmids to introduce normal genes into
        cells that contain abnormal genes.




       Insight
Why Anaerobes Die in the Presence of Oxygen
When molecular oxygen (O2) is reduced (i.e., when O2 gains electrons; as in certain oxidation–
reduction reactions), extremely reactive substances are produced (as shown in the following
equations).

                                O2 + e–  –O2 (superoxide anion)

                              O2 + 2e–  H2O2 (hydrogen peroxide)

                            O2 + 3e–  H2O + OH– (hydroxyl radical)

         These reduction products (superoxide anion, hydrogen peroxide, and hydroxyl radicals)
are capable of causing severe damage to enzymes and cell membranes; they are potentially lethal
to cells. To survive in the presence of oxygen, organisms must possess enzymes (e.g., superoxide
dismutase and catalase) that can neutralize these toxic substances. Obligate anaerobes are killed
in the presence of oxygen because they lack one or more of these enzymes. Aerotolerant
anaerobes produce these enzymes, but not in high enough concentrations to enable the organisms
to survive in high concentrations of oxygen.
Chapter 7: Microbial Physiology and Genetics         6




Genetically Engineered Bacteria and Yeasts
The term genetic engineering refers to the manufacture and manipulation of genetic material in
vitro (in the laboratory). Genetic engineering has been possible only since the late 1960s, when a
scientist named Paul Berg demonstrated that fragments of human or animal DNA can be attached
to bacterial DNA. Such a hybrid DNA molecule is referred to as recombinant DNA. When a
molecule of recombinant DNA is inserted into a bacterial cell, the bacterium is able to produce
the gene product, usually a protein. Thus, microorganisms (primarily bacteria) can be genetically
engineered to produce substances (gene products) that they would not normally manufacture.
Paul Berg won a Nobel Prize in 1980 for his pioneering genetic engineering experiments.
        Molecules of self-replicating, extrachromosomal DNA, called plasmids, are frequently
used in genetic engineering and are referred to as vectors. A particular gene of interest is first
inserted into the vector DNA, forming a molecule of recombinant DNA. The recombinant DNA
is then inserted into or taken up by a bacterial cell. The cell is next allowed to multiply, creating
many genetically identical bacteria (clones), each of which is capable of producing the gene
product. From the clone culture, a genetic engineer may then remove (―harvest‖) the gene
product.
        The Gram-negative bacillus, Escherichia coli, has often been used because it can be
easily grown in the laboratory, has a relatively short generation time (about 20 minutes under
ideal conditions), and its genetics are well understood by researchers. A Gram-positive bacterium
(Bacillus subtilis), a yeast (Saccharomyces cerevisiae), and cultured plant and mammalian cells
have also been used by genetic engineers to produce desired gene products.
         An example of a product produced by genetic engineering is insulin, a hormone
produced in E. coli cells and used to treat diabetic patients. Human growth hormone
(somatotropin), bovine growth hormone (BGH), porcine growth hormone (PGH), somatostatin (a
hormone used to limit growth), tissue growth factors, clotting factors, and interferon are also
produced by genetically engineered E. coli. Genetically engineered bacteria are being used to
produce industrial enzymes, citric acid, and ethanol, and to degrade pollutants and toxic wastes.
The hepatitis B vaccine that is administered to healthcare workers is produced by a genetically
engineered yeast, called Saccharomyces cerevisiae.
        New uses for recombinant DNA and genetic engineering are being discovered every day,
causing profound changes in medicine, agriculture, and other areas of science.




      Increase Your Knowledge
1.       A Closer Look at Transduction. There are actually two types of transduction:
specialized and generalized. The explanation in Chapter 7 describes specialized transduction, in
which the infecting phage integrates into the bacterial chromosome or a plasmid. As the virus
genome breaks away to replicate and produce more viruses, it carries one or more bacterial genes
with it to the newly infected cell. In this way, genetic capabilities involving the fermentation of
certain sugars, antibiotic resistance, and other phenotypic characteristics can be transduced to
other bacteria. This process has been shown in the laboratory (in vitro) to occur in species of
Chapter 7: Microbial Physiology and Genetics         7




Bacillus, Pseudomonas, Haemophilus, Salmonella, and Escherichia, and it is assumed to occur
in nature. In generalized transduction, the bacteriophage is a virulent lytic phage that does not
incorporate into the bacterial genome or plasmid. Rather, it picks up fragments of bacterial DNA
during the assembly of new virus particles and carries these bacterial genes to other cells that the
new viruses infect. Generalized transduction has been observed in species of Streptococcus,
Staphylococcus, and Salmonella, and in Vibrio cholerae.

2.       A Closer Look at Fertility Factors. Bacteria possessing F+ or Hfr+ genes have the
ability to produce sex pili and become donor cells. If the fertility factor is on a plasmid, it is
called an F+ gene, whereas if it is incorporated into the chromosome, it is referred to as an HFr+
gene. A complete copy of the F plasmid (the plasmid containing the F+ gene) usually moves to
the recipient (F-) cell; therefore, the recipient cell usually becomes F+ (i.e., the recipient cell
becomes capable of producing a sex pilus and becoming a donor cell). On the other hand, the
recipient cell usually receives only a portion of the chromosome from an HFr+ cell, and that
portion does not include the HFr+ gene; therefore, in this case, the recipient cell remains Hfr–,
does not produce a sex pilus, and cannot become a donor cell.

3.     Learn more about enzymes at:
       www.northland.cc.mn.us/biology/biology1111/animations/enzyme.swf

4.     View the following animations on the TCA Cycle and Electron Transport:

       Click here to view TCA Cycle

       Click here to view Electron Transport

5.     Learn more about transduction, conjugation and transformation at:
       www.emunix.emich.edu/~rwinning/genetics/bactrec.htm


6.     Try your hand at some advanced microbial genetic problems at:
       www.sci.sdsu.edu/~smaloy/MicrobialGenetics/problems/

7.     There are many good videos on genetics on YouTube. For example, try this one:
       www.youtube.com/watch?v=t4i0Q_irM8o

8.     The HowStuffWorks website also has many good videos on bacterial mutations and
genetics. Check it out at:
       videos.howstuffworks.com/discovery/28648-assignment-discovery-bacteria-mutations-
       video.htm

9.     Learn more about gene therapy at:
       www.ornl.gov/sci/techresources/Human_Genome/medicine/genetherapy.shtml
Chapter 7: Microbial Physiology and Genetics        8




      Critical Thinking
1.     What are some possible reasons why an obligate anaerobe is unable to live in the
       presence of oxygen?

2.     Assume that you are a microbiologist who has been doing research on a penicillin-
       sensitive strain of Staphylococcus aureus for many months. One day you discover that
       the organism is now resistant to penicillin. You know that it has not come in contact with
       any other species of bacteria, nor has it come in contact with the DNA from any other
       species of bacteria. What are two possible explanations for its sudden change from
       penicillin susceptibility to penicillin resistance?

3.     Several products were mentioned in this chapter that are being produced by genetically
       engineered bacteria and yeasts. Using the Internet, can you find others?




Answers to the Chapter 7 Self-Assessment Exercises in the Text
 1.    C
 2.    A
 3.    D
 4.    A
 5.    C
 6.    D
 7.    D
 8.    B
 9.    C
10.    A




      Additional Chapter 7 Self-Assessment Exercises
(Note: Don’t peek at the answers before you attempt to solve these self-assessment exercises.)
Chapter 7: Microbial Physiology and Genetics       9




Matching Questions
A.   autotrophs             _____ 1.     _______________ are chemotrophs that
B.   heterotrophs                        use inorganic chemicals as their energy
C.   lithotrophs                         source.
D.   organotrophs
E.   phototrophs            _____ 2.     Organisms that use organic compounds
                                         as their source of carbon are called
                                         _______________.

                            _____ 3.     Organisms that use organic compounds
                                         as their energy source are called
                                         _______________.

                            _____ 4.     Organisms that use carbon dioxide as
                                         their source of carbon are called
                                         _______________.

                            _____ 5.     Organisms that use light as their energy
                                         source are called _______________.

A.   conjugation            _____ 6.     In _______________, bacteria acquire
B.   lysogenic conversion                new genetic information in the form of
C.   mutation                            viral genes.
D.   transduction
E.   transformation         _____ 7.     Oswald Avery and his colleagues
                                         discovered that DNA is the hereditary
                                         molecule while performing
                                         _______________ experiments with
                                         Streptococcus pneumoniae.

                            _____ 8.     In _______________, bacteria acquire
                                         new genetic information as a result of
                                         absorbing pieces of naked DNA from
                                         their environment.

                            _____ 9.     In _______________, genetic
                                         information is passed from one bacterial
                                         cell to another via a hollow sex pilus.

                            _____ 10.    In _______________, bacteria acquire
                                         new genetic information when
                                         bacteriophages inject bacterial genes.

True/False Questions
Chapter 7: Microbial Physiology and Genetics          10




_____ 1.    Dehydration synthesis reactions always involve the removal of a molecule of
            water.

_____ 2.    The biosynthesis of polysaccharides, polypeptides, and nucleic acids are examples
            of catabolic reactions.

_____ 3.    Oxidation–reduction reactions are paired reactions that involve the transfer of
            electrons.

_____ 4.    Breaking a disaccharide down into its two monosaccharide components is an
            example of a hydrolysis reaction.

_____ 5.    Anabolic reactions are a cell’s major source of energy.

_____ 6.    The majority of energy produced in aerobic respiration is produced by the Krebs
            cycle.

_____ 7.    In glycolysis, a 6-carbon glucose molecule is broken down into two 3-carbon
            molecules of pyruvic acid.

_____ 8.    Aerobic respiration is a more efficient method of breaking down glucose than is
            fermentation.

_____ 9.    Virulent bacteriophages are responsible for lysogenic conversion.

_____ 10.   Mutations are always harmful.




Answers to the Additional Chapter 7 Self-Assessment Exercises
Chapter 7: Microbial Physiology and Genetics   11




Matching Questions

 1.   C
 2.   B
 3.   D
 4.   A
 5.   E
 6.   B
 7.   E
 8.   E
 9.   A
10.   D


True/False Questions

 1.   True
 2.   False (these are examples of anabolic reactions)
 3.   True
 4.   True
 5.   False (catabolic reactions are a cell’s major source of energy)
 6.   False (the majority of the energy is produced by the electron transport chain)
 7.   True
 8.   True
 9.   False (temperate bacteriophages are responsible for lysogenic conversion)
10.   False (mutations may be harmful, beneficial, or ―silent‖)

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Sr chapter007

  • 1. Chapter 7: Microbial Physiology and Genetics 1 Chapter 7 Microbial Physiology and Genetics Primary Objectives of the Chapter Chapter 7 introduces aspects of microbial physiology, such as enzymes, metabolism (catabolic and anabolic reactions), oxidation–reduction reactions, biochemical pathways, aerobic respiration, and fermentation. Microbial genetic topics discussed in Chapter 7 include mutations, the various ways in which bacteria acquire new genetic information (lysogenic conversion, transduction, transformation, conjugation), genetic engineering, and gene therapy. The information in Chapter 7 is considered essential in an introductory microbiology course. Terms Introduced in This Chapter After reading Chapter 7, you should be familiar with the following terms. These terms are defined in Chapter 7 and in the Glossary. Adenosine triphosphate (ATP) Ames test Anabolic reactions Anabolism Autotroph Beneficial mutation Catabolic reactions Catabolism Chemoautotroph Chemoheterotroph Chemolithotroph Chemoorganotroph Chemosynthesis Chemotroph Competence Competent bacteria Dehydrogenation reactions Ecology Ecosystem Electron transport chain Endoenzyme Episome Essential nutrients Exoenzyme Fermentation Gene therapy
  • 2. Chapter 7: Microbial Physiology and Genetics 2 Genetics Glycolysis Harmful mutation Heterotroph Krebs cycle Lethal mutation Lysogenic bacterium Lysogenic conversion Lysogeny Metabolic reactions Metabolite Microbial physiology Mutagen Mutant Mutation Oxidation Oxidation–reduction reactions Phenotype Photoautotroph Photoheterotroph Phototroph Prophage R-factor Reduction Silent mutation Transduction Transformation Review of Key Points  Microbial physiology is the study of the life processes of microorganisms.  Scientists have learned a great deal about cells—including human cells—by studying the nutritional needs of bacteria, their metabolic pathways, and why they live, grow, multiply, or die under certain conditions.  All living organisms require sources of energy and carbon so that they can produce the molecules necessary for life. In addition, organisms must be provided with certain materials (called essential nutrients) that they themselves are unable to synthesize, but are required for survival; these essential nutrients vary from species to species.  The energy source for certain organisms (called phototrophs) is light and for other organisms (called chemotrophs) is organic or inorganic chemicals.  Chemolithotrophs and chemoorganotrophs are subcategories of chemotrophs. Chemolithotrophs (or simply lithotrophs) use inorganic chemicals as an energy source,
  • 3. Chapter 7: Microbial Physiology and Genetics 3 whereas chemoorganotrophs (or simply organotrophs) use organic chemicals as an energy source.  An organism’s carbon source may be CO2 (in which case the organism is called an autotroph) or other organic compounds (in which case the organism is called a heterotroph). Humans, animals, protozoa, and fungi are heterotrophs, as are most bacteria.  Interrelationships among the different nutritional types are of prime importance in the functioning of the ecosystem. Phototrophs (plants, algae, and certain bacteria) are the producers of food and oxygen for the chemoheterotrophs (animals). Dead plants and animals are recycled by the chemoheterotrophic saprophytic decomposers (certain fungi and bacteria) into nutrients for phototrophs and chemotrophs.  Metabolism refers to all of the chemical reactions (metabolic reactions) that occur within a living cell, including the production of energy and the synthesis of new molecules.  Metabolic reactions include catabolic reactions and anabolic reactions. Catabolic reactions (also called degradative reactions) involve the breaking of chemical bonds and the release of energy. Anabolic reactions (also called biosynthetic reactions) require energy because they involve the formation of chemical bonds.  Most metabolic reactions are regulated by enzymes.  Enzymes are biologic molecules (proteins) that serve as catalysts to control the rate of metabolic reactions. The enzymes produced by any particular cell are governed by the genotype of that cell, and the presence or absence of any particular enzyme is part of the phenotype of that cell.  The substance upon which an enzyme acts is known at that enzyme’s substrate.  All the enzymes that a cell is capable of producing need not be present in the cell at a given moment in time. They are produced to meet the metabolic needs of the cell, as determined by the internal and external environments.  Endoenzymes are enzmes that remain within the cell that produced them, whereas exoenzymes are enzymes that leave the cell to catalyze reactions outside of the cell.  Apoenzymes are proteins that are unable to catalyze reactions on their own. To catalyze reactions, apoenzymes must first link up with a cofactor (either a mineral ion or a coenzyme).  An enzyme operates at peak efficiency within a particular pH and temperature range and when an appropriate concentration of the substrate for that enzyme exists. If the environment is too acidic, basic, hot, or cold, or contains too much or too little substrate, the enzyme will not operate at peak efficiency and the reaction will not proceed at its maximum rate.  Catabolic reactions involve the breaking of chemical bonds and the release of energy. Anabolic reactions involve the formation of bonds, which requires energy.  Adenosine triphosphate (ATP) is the principal energy-storing or energy-carrying molecule in the cell. Should a cell require energy, one of the high-energy bonds in an ATP molecule can be broken, producing energy, an ADP molecule, and a free phosphate. The energy can then be used for growth, reproduction, active transport of substances across membranes, sporulation, movement, anabolic reactions, and other energy- requiring activities.
  • 4. Chapter 7: Microbial Physiology and Genetics 4  Nutrients should be thought of as energy sources, and chemical bonds should be thought of as stored energy.  A common pathway by which bacteria catabolize glucose is aerobic respiration, which consists of three phases: glycolysis, the Krebs cycle, and the electron transport chain. Most of the energy that is produced by aerobic respiration is produced by the electron transport chain. The breakdown of one molecule of glucose by aerobic respiration yields either 36 ATP molecules (procaryotic cells) or 38 ATP molecules (eucaryotic cells).  Aerobes and facultative anaerobes are able to produce more energy than anaerobes, because they can catabolize glucose molecules via aerobic pathways. Anaerobes must catabolize glucose by fermentation, a relatively inefficient method that yields only two ATP molecules from a molecule of glucose.  Oxygen does not participate in fermentation reactions.  Oxidation reactions involve the loss of an electron, whereas reduction reactions involve the gain of an electron.  Phototrophic organisms (algae, plants, and photosynthetic bacteria) derive their energy from the sun by photosynthesis. Chemosynthetic organisms use a chemical source of energy and raw materials to synthesize metabolites and macromolecules for growth and function of the organisms.  As with humans, animals, and plants, the genetics of microbes involves DNA, genes, the genetic code, chromosomes, DNA replication, transcription, and translation—all part of molecular genetics.  An organism’s genotype (or genome) is its complete collection of genes, whereas an organism’s phenotype is all the physical traits, attributes, or characteristics of the organism.  Genes direct all functions of the cell, providing it with its own particular traits and individuality. An organism’s phenotype is the manifestation of that organism’s genotype.  Constitutive genes are expressed at all times, whereas inducible genes are expressed only when the products that they code for (gene products) are needed.  The base sequence of any gene on a chromosome may be altered accidentally in many ways, resulting in a mutation. Mutations are expressed not only in the cell in which the mutation occurred, but in subsequent generations as well. The altered genetic code will result in an altered protein, which could affect any of a number of different phenotypic characteristics (e.g., changes in colony characteristics, cell shape, biochemical activities, nutritional needs, antigenic sites, virulence, pathogenicity, drug resistance). Mutant bacteria are used in genetic and medical research and the production of vaccines.  Mutations may be beneficial, harmful, or of no consequence to the cell or organism containing the mutation. Those of no consequence are called silent or neutral mutations.  Beneficial mutations are of benefit to an organism, whereas harmful mutations result in the production of structurally altered proteins (often, nonfunctional enzymes). Some harmful mutations are lethal to the organism.  Physical or chemical agents that cause an increased mutation rate are called mutagens.
  • 5. Chapter 7: Microbial Physiology and Genetics 5  In addition to mutations, genetic changes in a bacterial cell may be the result of lysogenic conversion, transduction, transformation, or conjugation, all of which occur in nature as well as in the laboratory.  Lysogenic conversion involves temperate bacteriophages. In lysogenic conversion, bacteria gain new genetic information in the form of viral genes.  Transduction also involves bacteriophages. In lysogenic conversion, bacteria gain new genetic information in the form of bacterial genes.  In transformation, a bacterial cell becomes genetically transformed following uptake of DNA fragments (―naked DNA‖) from the environment.  Conjugation involves the transfer of genetic material (usually a plasmid) from a donor cell to a recipient cell through a hollow sex pilus.  A plasmid that contains multiple genes for antibiotic resistance is called a resistance factor or R-factor.  The field of genetic engineering involves the introduction of new genes into cells. When a cell receives a new gene, it can produce the gene product that is coded for by that gene. Genetically engineered bacteria are used to produce products such as insulin, interferon, human growth hormone, and materials for use as vaccines.  Gene therapy involves the use of viruses and plasmids to introduce normal genes into cells that contain abnormal genes. Insight Why Anaerobes Die in the Presence of Oxygen When molecular oxygen (O2) is reduced (i.e., when O2 gains electrons; as in certain oxidation– reduction reactions), extremely reactive substances are produced (as shown in the following equations). O2 + e–  –O2 (superoxide anion) O2 + 2e–  H2O2 (hydrogen peroxide) O2 + 3e–  H2O + OH– (hydroxyl radical) These reduction products (superoxide anion, hydrogen peroxide, and hydroxyl radicals) are capable of causing severe damage to enzymes and cell membranes; they are potentially lethal to cells. To survive in the presence of oxygen, organisms must possess enzymes (e.g., superoxide dismutase and catalase) that can neutralize these toxic substances. Obligate anaerobes are killed in the presence of oxygen because they lack one or more of these enzymes. Aerotolerant anaerobes produce these enzymes, but not in high enough concentrations to enable the organisms to survive in high concentrations of oxygen.
  • 6. Chapter 7: Microbial Physiology and Genetics 6 Genetically Engineered Bacteria and Yeasts The term genetic engineering refers to the manufacture and manipulation of genetic material in vitro (in the laboratory). Genetic engineering has been possible only since the late 1960s, when a scientist named Paul Berg demonstrated that fragments of human or animal DNA can be attached to bacterial DNA. Such a hybrid DNA molecule is referred to as recombinant DNA. When a molecule of recombinant DNA is inserted into a bacterial cell, the bacterium is able to produce the gene product, usually a protein. Thus, microorganisms (primarily bacteria) can be genetically engineered to produce substances (gene products) that they would not normally manufacture. Paul Berg won a Nobel Prize in 1980 for his pioneering genetic engineering experiments. Molecules of self-replicating, extrachromosomal DNA, called plasmids, are frequently used in genetic engineering and are referred to as vectors. A particular gene of interest is first inserted into the vector DNA, forming a molecule of recombinant DNA. The recombinant DNA is then inserted into or taken up by a bacterial cell. The cell is next allowed to multiply, creating many genetically identical bacteria (clones), each of which is capable of producing the gene product. From the clone culture, a genetic engineer may then remove (―harvest‖) the gene product. The Gram-negative bacillus, Escherichia coli, has often been used because it can be easily grown in the laboratory, has a relatively short generation time (about 20 minutes under ideal conditions), and its genetics are well understood by researchers. A Gram-positive bacterium (Bacillus subtilis), a yeast (Saccharomyces cerevisiae), and cultured plant and mammalian cells have also been used by genetic engineers to produce desired gene products. An example of a product produced by genetic engineering is insulin, a hormone produced in E. coli cells and used to treat diabetic patients. Human growth hormone (somatotropin), bovine growth hormone (BGH), porcine growth hormone (PGH), somatostatin (a hormone used to limit growth), tissue growth factors, clotting factors, and interferon are also produced by genetically engineered E. coli. Genetically engineered bacteria are being used to produce industrial enzymes, citric acid, and ethanol, and to degrade pollutants and toxic wastes. The hepatitis B vaccine that is administered to healthcare workers is produced by a genetically engineered yeast, called Saccharomyces cerevisiae. New uses for recombinant DNA and genetic engineering are being discovered every day, causing profound changes in medicine, agriculture, and other areas of science. Increase Your Knowledge 1. A Closer Look at Transduction. There are actually two types of transduction: specialized and generalized. The explanation in Chapter 7 describes specialized transduction, in which the infecting phage integrates into the bacterial chromosome or a plasmid. As the virus genome breaks away to replicate and produce more viruses, it carries one or more bacterial genes with it to the newly infected cell. In this way, genetic capabilities involving the fermentation of certain sugars, antibiotic resistance, and other phenotypic characteristics can be transduced to other bacteria. This process has been shown in the laboratory (in vitro) to occur in species of
  • 7. Chapter 7: Microbial Physiology and Genetics 7 Bacillus, Pseudomonas, Haemophilus, Salmonella, and Escherichia, and it is assumed to occur in nature. In generalized transduction, the bacteriophage is a virulent lytic phage that does not incorporate into the bacterial genome or plasmid. Rather, it picks up fragments of bacterial DNA during the assembly of new virus particles and carries these bacterial genes to other cells that the new viruses infect. Generalized transduction has been observed in species of Streptococcus, Staphylococcus, and Salmonella, and in Vibrio cholerae. 2. A Closer Look at Fertility Factors. Bacteria possessing F+ or Hfr+ genes have the ability to produce sex pili and become donor cells. If the fertility factor is on a plasmid, it is called an F+ gene, whereas if it is incorporated into the chromosome, it is referred to as an HFr+ gene. A complete copy of the F plasmid (the plasmid containing the F+ gene) usually moves to the recipient (F-) cell; therefore, the recipient cell usually becomes F+ (i.e., the recipient cell becomes capable of producing a sex pilus and becoming a donor cell). On the other hand, the recipient cell usually receives only a portion of the chromosome from an HFr+ cell, and that portion does not include the HFr+ gene; therefore, in this case, the recipient cell remains Hfr–, does not produce a sex pilus, and cannot become a donor cell. 3. Learn more about enzymes at: www.northland.cc.mn.us/biology/biology1111/animations/enzyme.swf 4. View the following animations on the TCA Cycle and Electron Transport: Click here to view TCA Cycle Click here to view Electron Transport 5. Learn more about transduction, conjugation and transformation at: www.emunix.emich.edu/~rwinning/genetics/bactrec.htm 6. Try your hand at some advanced microbial genetic problems at: www.sci.sdsu.edu/~smaloy/MicrobialGenetics/problems/ 7. There are many good videos on genetics on YouTube. For example, try this one: www.youtube.com/watch?v=t4i0Q_irM8o 8. The HowStuffWorks website also has many good videos on bacterial mutations and genetics. Check it out at: videos.howstuffworks.com/discovery/28648-assignment-discovery-bacteria-mutations- video.htm 9. Learn more about gene therapy at: www.ornl.gov/sci/techresources/Human_Genome/medicine/genetherapy.shtml
  • 8. Chapter 7: Microbial Physiology and Genetics 8 Critical Thinking 1. What are some possible reasons why an obligate anaerobe is unable to live in the presence of oxygen? 2. Assume that you are a microbiologist who has been doing research on a penicillin- sensitive strain of Staphylococcus aureus for many months. One day you discover that the organism is now resistant to penicillin. You know that it has not come in contact with any other species of bacteria, nor has it come in contact with the DNA from any other species of bacteria. What are two possible explanations for its sudden change from penicillin susceptibility to penicillin resistance? 3. Several products were mentioned in this chapter that are being produced by genetically engineered bacteria and yeasts. Using the Internet, can you find others? Answers to the Chapter 7 Self-Assessment Exercises in the Text 1. C 2. A 3. D 4. A 5. C 6. D 7. D 8. B 9. C 10. A Additional Chapter 7 Self-Assessment Exercises (Note: Don’t peek at the answers before you attempt to solve these self-assessment exercises.)
  • 9. Chapter 7: Microbial Physiology and Genetics 9 Matching Questions A. autotrophs _____ 1. _______________ are chemotrophs that B. heterotrophs use inorganic chemicals as their energy C. lithotrophs source. D. organotrophs E. phototrophs _____ 2. Organisms that use organic compounds as their source of carbon are called _______________. _____ 3. Organisms that use organic compounds as their energy source are called _______________. _____ 4. Organisms that use carbon dioxide as their source of carbon are called _______________. _____ 5. Organisms that use light as their energy source are called _______________. A. conjugation _____ 6. In _______________, bacteria acquire B. lysogenic conversion new genetic information in the form of C. mutation viral genes. D. transduction E. transformation _____ 7. Oswald Avery and his colleagues discovered that DNA is the hereditary molecule while performing _______________ experiments with Streptococcus pneumoniae. _____ 8. In _______________, bacteria acquire new genetic information as a result of absorbing pieces of naked DNA from their environment. _____ 9. In _______________, genetic information is passed from one bacterial cell to another via a hollow sex pilus. _____ 10. In _______________, bacteria acquire new genetic information when bacteriophages inject bacterial genes. True/False Questions
  • 10. Chapter 7: Microbial Physiology and Genetics 10 _____ 1. Dehydration synthesis reactions always involve the removal of a molecule of water. _____ 2. The biosynthesis of polysaccharides, polypeptides, and nucleic acids are examples of catabolic reactions. _____ 3. Oxidation–reduction reactions are paired reactions that involve the transfer of electrons. _____ 4. Breaking a disaccharide down into its two monosaccharide components is an example of a hydrolysis reaction. _____ 5. Anabolic reactions are a cell’s major source of energy. _____ 6. The majority of energy produced in aerobic respiration is produced by the Krebs cycle. _____ 7. In glycolysis, a 6-carbon glucose molecule is broken down into two 3-carbon molecules of pyruvic acid. _____ 8. Aerobic respiration is a more efficient method of breaking down glucose than is fermentation. _____ 9. Virulent bacteriophages are responsible for lysogenic conversion. _____ 10. Mutations are always harmful. Answers to the Additional Chapter 7 Self-Assessment Exercises
  • 11. Chapter 7: Microbial Physiology and Genetics 11 Matching Questions 1. C 2. B 3. D 4. A 5. E 6. B 7. E 8. E 9. A 10. D True/False Questions 1. True 2. False (these are examples of anabolic reactions) 3. True 4. True 5. False (catabolic reactions are a cell’s major source of energy) 6. False (the majority of the energy is produced by the electron transport chain) 7. True 8. True 9. False (temperate bacteriophages are responsible for lysogenic conversion) 10. False (mutations may be harmful, beneficial, or ―silent‖)