Chapter5 Microbial Nutrition

PA RT II Microbial Nutrition, Growth, and Control

5. Microbial Nutrition

• Nutrient requirements

• Nutritional types of microorganisms

• Uptake of Nutrients by the Cell

• Culture Medium

• Isolation of Pure Cultures

Outline:

Concepts1. Microorganisms require about 10 elements in large quantities, in part

because they are used to construct carbohydrates, lipids, proteins, and nucleic acids. Several other elements are needed in very small amounts and are parts of enzymes and cofactors.

2. All microorganisms can be placed in one of a few nutritional categories on the basis of their requirements for carbon, energy, and hydrogen atoms or electrons.

3. Nutrient molecules frequently cannot cross selectively permeable plasma membranes through passive diffusion. They must be transported by one of three major mechanisms involving the use of membrane carrier proteins. Eucaryotic microorganisms also employ endocytosis for nutrient uptake.

4. Culture media are needed to grow microorganisms in the laboratory and to carry out specialized procedures like microbial identification, water and food analysis, and the isolation of articular microorganisms. Many different media are available for these and other purposes.

5. Pure cultures can be obtained through the use of spread plates, streak plates, or pour plates and are required for the careful study of an individual microbial species.

Macronutrients

• 95% or more of cell dry weight is made up of a few major elements: carbon, oxygen, hydrogen, nitrogen, sulfur, phosphorus, potassium, calcium, magnesium and iron.

• The first six ( C, H, O, N, P and S) are components of carbonhadrates, lipids, proteins and nucleic acids

Trace Elements

Microbes require very small amounts of other mineral elements, such as iron, copper, molybdenum, and zinc; these are referred to as trace elements. Most are essential for activity of certain enzymes, usually as cofactors.

Growth Factors

Amino acids are needed for protein synthesis,

purines and pyrimidines for nucleic acid synthesis.

Vitamins are small organic molecules that usually make up all or part enzyme cofactors, and only very small amounts are required for growth.

(1)amino acids, (2) purines and pyrimidines, (3) vitamins

Major nutritional type

Sources of energy,

hydrogen/electrons,

and carbon

Representative microorganisms

Photoautotroph

(Photolithotroph)

Light energy, inorganic hydrogen/electron(H/e-) donor, CO2 carbon source

Algae, Purple and green bacteria, Cyanobacteria

Photoheterotroph

(Photoorganotroph)

Light energy, inorganic H/e- donor,

Organic carbon source

Purple nonsulfur bacteria,

Green sulfur bacteria

Chemoautotroph

(Chemolithotroph)

Chemical energy source (inorganic), Inorganic H/e- donor, CO2 carbon source

Sulfur-oxdizing bacteria, Hydrogen bacteria,

Nitrifying bacteria

Chemoheterotroph

(Chenoorganotroph)

Chemical energy source (organic), Organic H/e- donor, Organic carbon source

Most bacteria, fungi, protozoa

Nutritional types of microorganisms

Algae, Cyanobacteria

CO2 + H2O Light + Chlorophyll

（ CH2O ） +O2

Purple and green bacteria

CO2 + 2H2S Light + bacteriochlorophyll

（ CH2O ） + H2

O + 2S

Purple nonsulfur bacteria (Rhodospirillum)

CO2 + 2CH3CHOHCH3 Light + bacteriochlorophyll （ CH2O ）

+ H2O + 2CH3COCH3

Photoautotroph:

Photoheterotroph:

Property cyanobacteria Green and purple bacteria

Purple nonsulfur bacteria

Photo - pigment Chlorophyll Bcteriochlorophyll Bcteriochlorophyll

O2 production Yes No No

Electron donors H2O H2, H2S, S H2, H2S, S

Carbon source CO2 CO2 Organic / CO2

Primary products

of energy conversion

ATP + NADPH ATP ATP

Properties of microbial photosynthetic systems

Chemoautotroph:

Nitrifying bacteria

2 NH4+ + 3 O2 2 NO2- + 2 H2O + 4 H+ + 132 Kcal

Bacteria Electron donor

Electron acceptor

Products

Alcaligens and Pseudomonas sp.

H2 O2 H2O

Nitrobacter NO2- O2 NO3

- , H2ONitrosomonas NH4

+ O2 NO2- , H2O

Desulfovibrio H2 SO4 2- H2O. H2S

Thiobacillus denitrificans S0. H2S NO3- SO4

2- , N2

Thiobacillus ferrooxidans Fe2+ O2 Fe3+ , H2O

are needed to grow microorganisms in the laboratory and to carry out specialized procedures like microbial identification, water and food analysis, and the isolation of particular microorganisms. A wide variety of media is available for these and other purposes.

Culture media

can be obtained through the use of spread plates, streak plates, or pour plates and are required for the careful study of an individual microbial species.

Pure cultures

Nutrient molecules frequently cannot cross selectively permeable plasma membranes through passive diffusion and must be transported by one of three major mechanisms involving the use of membrane carrier proteins.

Uptake of nutrients

1, Phagocytosis – Protozoa

2, Permeability absorption – Most microorganisms

• passive transport , simple diffusion• facilitated diffusion• active transport• group translocation

A few substances, such as glycerol, can cross the plasma membrane by passive diffusion. Passive diffusion is the process in which molecules move from a region of higher concentration to one of lower concentration as a result of random thermal agitation.

passive diffusion

The rate of diffusion across selectively permeable membranes is greatly increased by the use of carrier proteins, sometimes called permeases, which are embedded in the plasina membrane. Since the diffusion process is aided by a carrier, it is called facilitated diffusion. The rate of facilitated diffusion increases with the concentratioti gradient much more rapidly and at lower concentrations of the diffusing molecule than that of passive diffusion

Facilitated diffusion

Active transport is the transport of solute molecules to higher concentrations, or against a concentration gradient, with the use of metabolic energy input.

Active transport

Group translocation

1, Phagocytosis – Protozoa

2, Permeability absorption – Most microorganisms

• passive transport , simple diffusion• facilitated diffusion• active transport• group translocation

A few substances, such as glycerol, can cross the plasma membrane by passive diffusion. Passive diffusion is the process in which molecules move from a region of higher concentration to one of lower concentration as a result of random thermal agitation.

passive diffusion

The rate of diffusion across selectively permeable membranes is greatly increased by the use of carrier proteins, sometimes called permeases, which are embedded in the plasina membrane. Since the diffusion process is aided by a carrier, it is called facilitated diffusion. The rate of facilitated diffusion increases with the concentration gradient much more rapidly and at lower concentrations of the diffusing molecule than that of passive diffusion

Facilitated diffusion

The membrane carrier can change conformation after binding an external molecule and subsequently release the molecule on the cell interior. It then returns to the outward oriented position and is ready to bind another solute molecule.

A model of facilitated diffusion

Because there is no energy input, molecules will continue to enter only as long as their concentration is greater on the outside.

Active transport is the transport of solute molecules to higher concentrations, or against a concentration gradient, with the use of metabolic energy input.

Active transport

The best-known group translocation system is the phosphoenolpyruvate: sugar phosphotransferase system (PTS), which transports a variety of sugars into procaryotic cells while Simultaneously phosphorylating them using phosphoenolpyruvate (PEP) as the phosphate donor.

Group translocationGroup translocation

PEP + sugar (outside) pyruvate + sugar-P (inside)

The phosphoenolpyruvate: sugar phosphotransferase system of E. coli. The following components are involved in the system: phosphoenolpyruvate, PEP; enzyme 1, E I; the low molecular weight heat-stable protein, HPr; enzyme 11, E II,- and enzyme III, E III.

Items Passive diffusion

Facilitated diffusion

Active transport

Group translocation

carrier proteins Non Yes Yes Yes

transport speed Slow Rapid Rapid Rapid

against gradient Non Non Yes Yes

transport molecules

No specificity Specificity Specificity Specificity

metabolic energy

No need Need Need Need

Solutes molecules

Not changed Changed Changed Changed

Simple comparison of transport systems

Mode of action of antibacterial antibiotics

Symbiosis of peptidoglycan

Questions

1. Throughout history, spices have been used as preservatives and to cover up the smell/taste of food that is slightly spoiled. The success of some spices led to a magical, ritualized use of many of them and possession of spices was often limited to priests or other powerful members of the community. a. Choose a spice and trace its use geographically and historically. What is its common-day use today? b. Spices grow and tend to be used predominantly in warmer climates. Explain.

2. Design an experiment to determine whether an antimicrobial agent is acting as a cidal or static agent. How would you determine whether an agent is suitable for use as an antiseptic rather than as a disinfectant?

3. Suppose that you are testing the effectiveness of disinfectants with the phenol coefficient test and obtained the following results.

Chapter 6 Microbial growth

• The Growth Curve

• Cell life cycle

• Measurement of Microbial Growth

• Measurement of Cell Mass

• Growth Yields and the Effects of a Limiting Nutrient

• The Continuous Culture of Microorganisms

• The Chemostat

• The Influence of Environmental Factors on Growth

Outline

1.   Growth is defined as an increase in cellular constituents and may result in an increase in an organism's size, population number, or both.

2.  When microorganisms are grown in a closed system, population growth remains exponential for only a few generations and then enters a stationary phase due to factors like nutrient limitation and waste accumulation. If a population is cultured in an open system with continual nutrient addition and waste removal, the exponential phase can be maintained for long periods.

Concepts

3. A wide variety of techniques can be used to study microbial growth by following changes in the total cell number, the population of viable microorganisms, or the cell mass.

4. Water availability, pH, temperature, oxygen concentration, pressure, radiation, and a number of other environmental factors influence microbial growth. Yet many microorganisms, and particularly bacteria, have managed to adapt and flourish under environmental extremes that would destroy most organisms.

Growth may be generally defined as a steady increase in all of the chemical components of an organism. Growth usually results in an increase in the size of a cell and frequently results in cell division

Growth definition:

G1 Primary growth phase of the cell during which cell enlargement occurs, a

gap phase separating cell growth from replication of the genome

S phase in which replication of the genome occurs

G2 Phase in which the cell prepares for separation of the replicated genomes,

this phase includes synthesis of microtubules and condensation of DNA to form coherent chromosomes, a gap phase separating chromosome replication from miosis.

M phase called miosis during which the microtubular apparatus is associated and subsequently used to pull apart the sister chromosomes.

Cell life cycle in Eukaryotic cells

G1 S G2 M

G1 R D

Eukaryotic cell:

Prokaryotic cell:

Most bacterial cells reproduce asexually by binary fision, a process in which a cell divides to produce two nearly equal-sized progeny cells. Binary fision involves three processes:

Increase in cell size (cell elongation),

DNA replication

Cell division

Binary fision

Growth curve of bacteriaGrowth curve of bacteria

1. Lag Phase

2. Exponential Phase

3.  Stationary Phase

4. Death Phase

Growth curve of bacteriaGrowth curve of bacteria

1. Lag Phase

2. Exponential Phase

3.  Stationary Phase

4. Death Phase

(a)      Lag phase: cells begin to synthesize inducible enzymes and use stored food reserves.

• (b)     Logarithmic growth phase: the rate of multiplication is constant.

• (c)      Stationary phase: death rate is equal to rate of increase.

• (d) Death phase: cells begin to die at a more rapid rate than that of reproduction.

Lag Phase

Stationary Phase

Death PhaseLogarithmic growth phase

The time required for a cell to divide (and its population to double) is called the generation time.

generation time

Suppose that a bacterial population increases from103 cells to 109 cells in 10 hours. Calculate the generation time.

Num

ber

of c

ells

Time

Nt = No x 2n

No = number of bacteria at beginning of time interval.

Nt = number of bacteria at end of any interval of time (t).

G = generation time

T = time , usually expressed in minutes

n = number of generation

G = t log2 / log Nt – log No

fermenter

Chemostat used for continuous cultures. Rate of growth can be controlled either by controlling the rate at which new medium enters the growth chamber or by limiting a required growth factor in the medium.

Continuous culture of microorganisms

Chemostat

Bacteria grow over a range of temperatures; they do not reproduce below the minimum growth temperattire nor above the maximum growth temperature. Within the temperature growth range there is an optimum growth temperature at which bacterial reproduction is fastest.

Effect of temperature on bacterial growth rate

Enzymes exhibit a Q10 so that within a suitable temperature range the rate of enzyme activity doubles for every 10' C rise in temperature.

Microorganisms are classified as psychrophiles, mesophiles.thermophiles, and extremethemophiles based on their optimal growth temperature.

Effect of oxygen concentration – reduction potential

Effect of oxygen concentration on the growth of various bacteria in tubes of solid medium

厌氧菌的培养

(a) Obligate aerobes-growth occurs only in the short distance to which the oxygen diffuses into the medium.

(b) Facultative anaerobes growth is best near the surface, where oxygen is available, but occurs throughout the tube.

(c) Obligate anaerobes-oxygen is toxic, and there is no growth near the surface.

(d) Aerotolerant anaerobes-growth occurs evenly throughout the tube but is not better at the surface because the organisms do not use oxygen.

(e) Microaerophiles, aerobic organisms that do not tolerate atmospheric concentrations of oxygen-growth occurs only in a narrow band of optimal oxygen concentration.

Effect of pH value on microbial growth

Bacteria: Neutral condition

Fungi: Acidic condition

Actinomycetes: Alkaline condition

Water activity

The water activity of a solution is 1/100 the relative humidity of the solution (when expressed as a percent), or it is equivalent to the ratio of the solution's vapor pressure to that of pure water.

aw = P solution / P water

Approximate lower aw limits for microbial growth:

0.90 – 1.00 for most bacteria, most algae and some fungi as Basidiomycetes,Mucor, Rhizopus.

0.75 for Halobacterium, Aspergillus…

0.60 for some saccharomyces species

If the concentration of solutes, such as sodium chloride, is higher in the surrounding medium (hypertonic), then water tends to leave the cell. The cell membrane shrinks away from the cell wall (an action called plasmolysis), and cell growth is inhibited.

Plasmolysis

Normal cell Plasmolyzed cell

Control of microbial growth

Definitions:

Sterilization – the process of destroying all forms of microbial life on an object or in a material.

Disinfection – the process of destroying vegetative pathogens but not necessary endospores.

Antisepsis – chemical disinfection of skin, mucous membranes or other living tissues