Chapter3
Microbial nutrition
1.Nutrient requirement
2.Nutritional types of microorganisms
3.Uptake of nutrients
4.Culture media
Microbial Growth Conditions
1,Macronutrients
2,Micronutrients
3,Growth factors
4,Environmental factors,temperature; pH; Oxygen et al.
Microorganisms require about ten elements in large
quantities,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.
Nutrient requirements
Microbial Nutrition
1,Macronutrients,required in large amounts,including,
carbon,oxygen,hydrogen,nitrogen,sulfur,phosphorus
(Components of carbohydrates,lipids,proteins,and nucleic
acids ); potassium,calcium,magnesium and iron (cations and
part of enzymes and cofactors),
2,Micronutrients,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,
Nutrients,Substances in the environment used by
organisms for catabolism and anabolism.
contaminants in water,glassware,and regular media
components often are adequate for growth.
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
Requirement for carbon,hydrogen and oxygen
Carbon sources,heterotrophs:,CHO”
autotrophs,CO2
Extraordinary flexibility:
No naturally occurring organic molecule cannot be
used by some microorganism,eg,Paraffin（石蜡）,rubber.
Omnivores,use over 100 different carbon compounds.
Fastidious,catabolize only a few carbon compound
Relatively indigestible human-made substances are
metabolized by complex populations of microorganisms.
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
(Chemoorganotroph)
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） +
H2O + 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-,H2O
Nitrosomonas NH4+ O2 NO2-,H2O
Desulfovibrio H2 SO4 2- H2O,H2S
Thiobacillus denitrificans S0,H2S NO3- SO4 2-,N2
Thiobacillus ferrooxidans Fe2+ O2 Fe3+,H2O
Nutritional types of microorganisms
? Phototrophs,use light as energy source.
? Chemotrophs,obtain energy from the
oxidation of chemical compounds.
? Lithotrophs,use reduced inorganic substances
as their electron source.
? Organotrophs,exteact electrons from organic
compounds.
Mixotrophic,many purple nonsulfur
bacteria
1,No oxygen,photoorganotrophic
heterotrophs
2,Normal oxygen,oxidize organic
molecules and function
chemotrophically.
3,Low oxygen,photosynthesis and
oxidative metabolism
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,H2O,O2
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,
no carrier protein;
no energy.
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 plasma membrane,Since the
diffusion process is aided by a carrier,it is called facilitated diffusion,
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
? higer con.? lower con.
? Facilitated diffusion,carrier protein,permeases.
? Each carrier is selective and will transport only
closely related solutes.
Seem not to be important in procaryotes,much
more prominent in Eucaryotic cells.
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.
?lower con.? higer con.
?Permeases,energy
Active transport
Proton gradients
? Symport,linked transport of
two substances in the same
direction.
? Antiport,linked transport of
two substances in the
opposite direction.
? Uniport,one substance enter
A process in which a molecule is transported
into the cell while being chemically altered.
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 translocation
PTS,sugar phosphortransferase system
PEP+sugar(outside)?pyruvate+sugar-P(inside)
The following components are involved in the system:
phosphoenolpyruvate,PEP;
EI (enzyme I),Hpr (the low molecular weight heat-stable protein),
cytoplasmic,common to all PTSs.
EII (enzyme II),
EIIa,cytoplasmic and soluble
EIIb,hydrophilic but frequently is attached to EIIc.
EIIc,a hydrophobic protein that is embedded in the
membrane.
Only specific sugars and varies with PTS.
The phosphoenolpyruvate,sugar phosphotransferase
system of E,coli,
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 No need Need Need
Solutes
molecules
Not changed Not changed Not changed Changed
Simple comparison of transport systems
Iron uptake
? Cytochromes and many enzymes
? Extreme insolubility of ferric iron(Fe3+) and its derivatives,
Difficult
? Siderophores,low M.W.,be able to complex with ferric
iron and supply it to the cell.
? Microorganisms secrete siderophores when little iron is
available in the medium.
iron-siderophore complex bind the receptor of cell surface,
Fe3+ release; complex enter by ABC transporter (ATP
binding casseset).
Siderophores (S)
Fe 2+/S
Receptor
Fe 2+/S
Culture media
A culture media is a solid or liquid
preparation used to grow,transport,and store
microorganisms,
Other functions:
Isolation and identification
The testing of antibiotic sensitivities
Water and food analysis
Industrial microbiology,et,al
Synthetic or defined media
? A medium in which all components are
known.
eg,Medium for E.coli (g/liter)
glucose 1.0; Na2HPO4 16.4; KH2PO4 1.5;
(NH4)2SO4 2.0; MgSO4 ·7H2O 200mg;
CaCl2 10mg; FeSO4 ·7H2O 0.5mg; Final
pH 6.8-7.0
Complex media
? Media that contain some ingredients of
unknown chemical composition like
peptones,meat extract,and yeast
extract.
? What is peptones? Protein hydrolysates
eg,LB medium,peptone; yeast extract;
sodium chloride; glucose.
Solid medium
? Liquid media+1.0-2.2% agar,common 1.5%.
? What is agar? Agar is a sulfated polymer composed
mainly of D-galactose,3,6-anhydro(无水 )-L-galactose,
and D-glucurouic acid（糖醛酸）,extracted from red
algae.
40-42℃ harden
80-90℃ melt
? Most microorganism cannot degrade it.
? Other,silica gel for autotrophic bacteria
Types of media
? Selective media,favor the growth of particular
microorganisms.
“加抑”, add componments which inhibite the
growth of unfavored bacteria.
“加富”, add components which promote the
growth of favored bacteria.
? Differential media,distinguish between different
groups of bacteria.
MacConkey agar,lactose,neutral red dye; lactose-
fermenting colonies appear pink to red in color.
What we have learned so far?
? Macro- and micronutrients? Defined medium and complex medium?
? Requirements for carbon,hydrogen,oxygen,nitrogen,phosphorus and sulfur?
? Nutritional types of microorganisms
? What are growth factors?
? What are facilitated diffusion,Active transport,Group translocation,endocytosis and their
difference?
? How do cell uptake iron?
? How to prepare a medium for cultivation microorganisms?
Microbial nutrition
1.Nutrient requirement
2.Nutritional types of microorganisms
3.Uptake of nutrients
4.Culture media
Microbial Growth Conditions
1,Macronutrients
2,Micronutrients
3,Growth factors
4,Environmental factors,temperature; pH; Oxygen et al.
Microorganisms require about ten elements in large
quantities,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.
Nutrient requirements
Microbial Nutrition
1,Macronutrients,required in large amounts,including,
carbon,oxygen,hydrogen,nitrogen,sulfur,phosphorus
(Components of carbohydrates,lipids,proteins,and nucleic
acids ); potassium,calcium,magnesium and iron (cations and
part of enzymes and cofactors),
2,Micronutrients,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,
Nutrients,Substances in the environment used by
organisms for catabolism and anabolism.
contaminants in water,glassware,and regular media
components often are adequate for growth.
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
Requirement for carbon,hydrogen and oxygen
Carbon sources,heterotrophs:,CHO”
autotrophs,CO2
Extraordinary flexibility:
No naturally occurring organic molecule cannot be
used by some microorganism,eg,Paraffin（石蜡）,rubber.
Omnivores,use over 100 different carbon compounds.
Fastidious,catabolize only a few carbon compound
Relatively indigestible human-made substances are
metabolized by complex populations of microorganisms.
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
(Chemoorganotroph)
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） +
H2O + 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-,H2O
Nitrosomonas NH4+ O2 NO2-,H2O
Desulfovibrio H2 SO4 2- H2O,H2S
Thiobacillus denitrificans S0,H2S NO3- SO4 2-,N2
Thiobacillus ferrooxidans Fe2+ O2 Fe3+,H2O
Nutritional types of microorganisms
? Phototrophs,use light as energy source.
? Chemotrophs,obtain energy from the
oxidation of chemical compounds.
? Lithotrophs,use reduced inorganic substances
as their electron source.
? Organotrophs,exteact electrons from organic
compounds.
Mixotrophic,many purple nonsulfur
bacteria
1,No oxygen,photoorganotrophic
heterotrophs
2,Normal oxygen,oxidize organic
molecules and function
chemotrophically.
3,Low oxygen,photosynthesis and
oxidative metabolism
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,H2O,O2
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,
no carrier protein;
no energy.
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 plasma membrane,Since the
diffusion process is aided by a carrier,it is called facilitated diffusion,
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
? higer con.? lower con.
? Facilitated diffusion,carrier protein,permeases.
? Each carrier is selective and will transport only
closely related solutes.
Seem not to be important in procaryotes,much
more prominent in Eucaryotic cells.
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.
?lower con.? higer con.
?Permeases,energy
Active transport
Proton gradients
? Symport,linked transport of
two substances in the same
direction.
? Antiport,linked transport of
two substances in the
opposite direction.
? Uniport,one substance enter
A process in which a molecule is transported
into the cell while being chemically altered.
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 translocation
PTS,sugar phosphortransferase system
PEP+sugar(outside)?pyruvate+sugar-P(inside)
The following components are involved in the system:
phosphoenolpyruvate,PEP;
EI (enzyme I),Hpr (the low molecular weight heat-stable protein),
cytoplasmic,common to all PTSs.
EII (enzyme II),
EIIa,cytoplasmic and soluble
EIIb,hydrophilic but frequently is attached to EIIc.
EIIc,a hydrophobic protein that is embedded in the
membrane.
Only specific sugars and varies with PTS.
The phosphoenolpyruvate,sugar phosphotransferase
system of E,coli,
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 No need Need Need
Solutes
molecules
Not changed Not changed Not changed Changed
Simple comparison of transport systems
Iron uptake
? Cytochromes and many enzymes
? Extreme insolubility of ferric iron(Fe3+) and its derivatives,
Difficult
? Siderophores,low M.W.,be able to complex with ferric
iron and supply it to the cell.
? Microorganisms secrete siderophores when little iron is
available in the medium.
iron-siderophore complex bind the receptor of cell surface,
Fe3+ release; complex enter by ABC transporter (ATP
binding casseset).
Siderophores (S)
Fe 2+/S
Receptor
Fe 2+/S
Culture media
A culture media is a solid or liquid
preparation used to grow,transport,and store
microorganisms,
Other functions:
Isolation and identification
The testing of antibiotic sensitivities
Water and food analysis
Industrial microbiology,et,al
Synthetic or defined media
? A medium in which all components are
known.
eg,Medium for E.coli (g/liter)
glucose 1.0; Na2HPO4 16.4; KH2PO4 1.5;
(NH4)2SO4 2.0; MgSO4 ·7H2O 200mg;
CaCl2 10mg; FeSO4 ·7H2O 0.5mg; Final
pH 6.8-7.0
Complex media
? Media that contain some ingredients of
unknown chemical composition like
peptones,meat extract,and yeast
extract.
? What is peptones? Protein hydrolysates
eg,LB medium,peptone; yeast extract;
sodium chloride; glucose.
Solid medium
? Liquid media+1.0-2.2% agar,common 1.5%.
? What is agar? Agar is a sulfated polymer composed
mainly of D-galactose,3,6-anhydro(无水 )-L-galactose,
and D-glucurouic acid（糖醛酸）,extracted from red
algae.
40-42℃ harden
80-90℃ melt
? Most microorganism cannot degrade it.
? Other,silica gel for autotrophic bacteria
Types of media
? Selective media,favor the growth of particular
microorganisms.
“加抑”, add componments which inhibite the
growth of unfavored bacteria.
“加富”, add components which promote the
growth of favored bacteria.
? Differential media,distinguish between different
groups of bacteria.
MacConkey agar,lactose,neutral red dye; lactose-
fermenting colonies appear pink to red in color.
What we have learned so far?
? Macro- and micronutrients? Defined medium and complex medium?
? Requirements for carbon,hydrogen,oxygen,nitrogen,phosphorus and sulfur?
? Nutritional types of microorganisms
? What are growth factors?
? What are facilitated diffusion,Active transport,Group translocation,endocytosis and their
difference?
? How do cell uptake iron?
? How to prepare a medium for cultivation microorganisms?
