Lect 7 - Nutrition

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Chapter 6
Microbial Nutrition
1
Elements of Life - 1
• macroelements (macronutrients)
– C, O, H, N, S, P
• found in organic molecules such as
proteins, lipids, carbohydrates, and
nucleic acids
– K, Ca, Mg, and Fe
• cations and serve in variety of roles
including enzymes, biosynthesis
– required in relatively large amounts
2
Table 6.1
3
Elements of Life - 2
• micronutrients (trace elements)
– Mn, Zn, Co, Mo, Ni, and Cu
– required in trace amounts
– often supplied in water or in media
components
– ubiquitous in nature
– serve as enzymes and cofactors
• some unique substances may be required
4
Carbon, Hydrogen, Oxygen,

and Electrons
• carbon is backbone of all organic
components present in cell
• hydrogen and oxygen are also found
in organic molecules
• electrons play a role in energy
production and reduction of CO2 to
form organic molecules
5
Requirements for Carbon,

Hydrogen, and Oxygen
• often satisfied together
– carbon source often provides H, O, and electrons
• heterotrophs
– use organic molecules as carbon sources which often
also serve as energy source
– can use a variety of carbon sources
• autotrophs
– use carbon dioxide as their sole or principal carbon
source
– must obtain energy from other sources
6
Table 6.2
7
Nutritional Types of Organisms

• based on energy source
– phototrophs use light
– chemotrophs obtain energy from
oxidation of chemical compounds
• based on electron source
– lithotrophs use reduced inorganic
substances
– organotrophs obtain electrons from
organic compounds
8
Classes of Major Nutritional

Types
• majority of microorganisms known
– photolithoautotrophs (photoautotrophs)
– chemoorganoheterotrophs
(chemoheterotrophs)
• majority of pathogens
• ecological importance
– photoorganoheterotrophs
– chemolithoautotrophs
– chemolithotrophs
9
Table 6.3
10
Figure 6.1
11
Microorganisms May Change

Nutritional Type
• some have great metabolic flexibility
based on environmental
requirements
• provides distinct advantage if
environmental conditions change
frequently
12
Figure 6.2
13
Requirements for Nitrogen,

Phosphorus, and Sulfur
• needed for synthesis of important
molecules (e.g., amino acids, nucleic acids)
• nitrogen supplied in numerous ways
• phosphorus usually supplied as inorganic
phosphate
• sulfur usually supplied as sulfate via
assimilatory sulfate reduction
14
Sources of Nitrogen
• organic molecules
• ammonia
• nitrate via assimilatory nitrate
reduction
• nitrogen gas via nitrogen fixation
15
Sources of Phosphorus and

Sulfur
• phosphorus
– most organisms use inorganic
phosphorus which is directly
incorporated into their cells
• sulfur
– most organisms use sulfate and reduce
it by assimilatory sulfate reduction
16
Growth Factors
• organic compounds
• essential cell components (or their
precursors) that the cell cannot
synthesize
• must be supplied by environment if
cell is to survive and reproduce
17
Classes of Growth Factors

• amino acids
– needed for protein synthesis
• purines and pyrimidines
– needed for nucleic acid synthesis
• vitamins
– function as enzyme cofactors??
• Heme
– Function in O2 binding
18
Table 6.4
19
Microbial Production of
Growth Factors
• microorganisms can synthesize
many growth factors
• large-scale industrial production of
growth factors, such as vitamins
20
Uptake of Nutrients
• microbes can only take in dissolved
particles across a selectively permeable
membrane
• some nutrients enter by passive diffusion
• microorganisms use transport
mechanisms
– facilitated diffusion – all microorganisms
– active transport – all microorganisms
– group translocation – Bacteria and Archaea
– endocytosis – Eukarya only
21
Passive Diffusion
• molecules move from region of
higher concentration to one of lower
concentration between the cell’s
interior and the exterior
• H2O, O2, and CO2 often move across
membranes this way
22
Facilitated Diffusion
• similar to passive diffusion
– movement of molecules is not energy
dependent
– direction of movement is from high
concentration to low concentration
– size of concentration gradient impacts
rate of uptake
23
Facilitated Diffusion…
• differs from passive diffusion
– uses membrane bound carrier molecules
(permeases)
– smaller concentration gradient is required
for significant uptake of molecules
– effectively transports glycerol, sugars, and
amino acids
• more prominent in eukaryotic cells than
in bacteria or archaea
24
•rate of facilitated
diffusion increases Figure 6.3
more rapidly and
at a lower
concentration
•diffusion rate
reaches a plateau
when carrier
becomes
saturated
25
Figure 6.4
26
Active Transport
• energy-dependent process
– ATP or proton motive force used
• move molecules against the gradient
• concentrates molecules inside cell
• involves carrier proteins (permeases)
– carrier saturation effect is observed at
high solute concentrations
27
ABC Transporters
• primary active transporters use ATP
• ATP-binding cassette transporters
• observed in Bacteria, Archaea, and
eukaryotes
• Consist of
– 2 hydrophobic membrane spanning domains
– 2 cytoplasmic associated ATP-binding domains
– Substrate binding domains
28
Figure 6.5
29
Secondary Active Transport

• major facilitator superfamily (MFS)
• use ion gradients to cotransport
substances
– protons
– symport – two substances both move in
the same direction
– antiport – two substances move in
opposite directions
30
Figure 6.6
31
Group Translocation
• energy dependent transport that
chemically modifies molecule as it is
brought into cell
• best known translocation system is
phosphoenolpyruvate: sugar
phosphotransferase system (PTS)
– many anaerobic bacteria transport sugars
while phosphorylating them using
phosphoenolpyruvate (PEP) as the
phosphate donor
32
Figure 6.7
33
Iron Uptake
• microorganisms
require iron
• ferric iron is very
insoluble so uptake is Figure 6.8
difficult
• microorganisms
secrete siderophores
to aid uptake
• siderophore
complexes with ferric
ion
• complex is then
transported into cell
34
Culture Media
• need to grow, transport, and store
microorganisms in the laboratory
• culture media is solid or liquid
preparation
• must contain all the nutrients required
by the organism for growth
• classification
– chemical constituents from which they are
made
– physical nature
– function
35
Table 6.5
36
Chemical and Physical

Types of Culture Media
Defined or synthetic
Complex
37
Defined or Synthetic Media

Table 6.6
• all components
and their
concentrations
are known
38
Complex Media
• contain some Table 6.7

ingredients of
unknown
composition
and/or
concentration
39
Some Media Components

• peptones
– protein hydrolysates prepared by partial
digestion of various protein sources
• extracts
– aqueous extracts, usually of beef or yeast
• agar
– sulfated polysaccharide used to solidify liquid
media; most microorganisms cannot degrade
it
40
Functional Types of Media
Supportive
Enriched
Selective
Differential
41
Functional Types of Media

• supportive or general purpose media
– support the growth of many
microorganisms
– e.g., tryptic soy agar
• enriched media
– general purpose media supplemented
by blood or other special nutrients
– e.g., blood agar
42
Figure 6.9
43
Selective Media
• favor the growth of some

microorganisms and inhibit growth
of others
• e.g., MacConkey agar
– selects for gram-negative bacteria
44
Differential Media
• distinguish between different groups of
microorganisms based on their biological
characteristics
• e.g., blood agar
– hemolytic versus nonhemolytic bacteria
• e.g., MacConkey agar
– lactose fermenters versus nonfermenters
45
Table 6.8
46
Isolation of Pure Cultures

• population of cells arising from a
single cell developed by Robert Koch
• allows for the study of single type of
microorganism in mixed culture
• spread plate, streak plate, and pour
plate are techniques used to isolate
pure cultures
47
The Streak Plate

• involves technique of spreading a
mixture of cells on an agar surface
so that individual cells are well
separated from each other
– involves use of bacteriological loop
• each cell can reproduce to form a
separate colony (visible growth or
cluster of microorganisms)
48
Figure 6.10
49
The Spread Plate and Pour

Plate
• spread plate
– small volume of diluted mixture containing
approximately 30–300 cells is transferred
– spread evenly over surface with a sterile bent rod
• pour plate
– sample is serially diluted
– diluted samples are mixed with liquid agar
– mixture of cells and agar are poured into sterile
culture dishes
• both may be used to determine the number of
viable microorganisms in an original sample
50
Figure 6.11
51
Figure 6.12
52
Microbial Growth on Solid

Surfaces
• colony characteristics that develop when
microorganisms are grown on agar
surfaces aid in identification
• microbial growth in biofilms is similar
• differences in growth rate from edges to
center is due to
– oxygen, nutrients, and toxic products
– cells may be dead in some areas
53
Figure 6.13
54

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Copyright © McGraw-Hill companies, Inc. Permission required for reproduction or display.

Carbon, Hydrogen, Oxygen,

Requirements for Carbon,

Nutritional Types of Organisms

Classes of Major Nutritional

Microorganisms May Change

Requirements for Nitrogen,

Sources of Phosphorus and

Classes of Growth Factors

Secondary Active Transport

Chemical and Physical

Defined or Synthetic Media

• contain some Table 6.7

Some Media Components

Functional Types of Media

Functional Types of Media

• favor the growth of some

Isolation of Pure Cultures

The Streak Plate

The Spread Plate and Pour

Microbial Growth on Solid

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