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.

Nutrient requirements
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.
 Microbial Nutrition
Nutrients: Substances in the environment used by
organisms for catabolism and anabolism.
1. Macronutrients: required in large amounts, including: c
arbon, oxygen, hydrogen, nitrogen, sulfur, phosphorus (Compone
nts of carbonhydrates, lipids, proteins, and mucleic acids ); potass
ium, calcium, magnesium and iron (cations and part of enzy
mes and cofactors).
2. Micronutrients: Microbes require very small amounts o
f other mineral elements, such as iron, copper, molybden
um, and zinc; these are referred to as trace elements. M
ost are essential for activity of certain enzymes, usually a
s cofactors.
contaminants in water, glassware, and regular media
components often are adequate for growth.
 Growth Factors

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

Amino acids are needed for protein synthesis,

purines and pyrimidines for nucleic acid synthesis.
Vitamins are small organic molecules that usually ma
ke up all or part enzyme cofactors, and only very s
mall amounts are required for growth.
Requirement for carbon, hydrogen and oxygen

Carbon sources: heterotrophs: “CHO”

autotrophs: CO2

Extraordinary flexibility:
No naturally occurring organic molecule cannot be us
ed by some microorganism. eg. Paraffin, rubber.
Omnivores: use over 100 different carbon compounds.
Fastidious: catabolize only a few carbon compound

Relatively indigestible human-made substances are m

etabolized by complex populations of microorganisms.
 Nutritional types of microorganisms
Sources of energy,
Major nutritional hydrogen/electrons, Representative
type and carbon microorganisms
Photoautotroph Light energy, inorganic Algae, Purple and green b
hydrogen/electron(H/e-) acteria, Cyanobacteria
(Photolithotroph) donor, CO2 carbon source

Photoheterotroph Light energy, inorganic Purple nonsulfur bacteria,

H/e- donor, Green sulfur bacteria
(Photoorganotroph)
Organic carbon source
Chemoautotroph Chemical energy source Sulfur-oxdizing bacteria,
(inorganic), Inorganic H/e- Hydrogen bacteria,
(Chemolithotroph) donor, CO2 carbon source Nitrifying bacteria

Chemoheterotroph Chemical energy source Most bacteria, fungi,

(organic), Organic H/e- protozoa
(Chenoorganotroph) donor, Organic carbon
source
 Photoautotroph:
Algae, Cyanobacteria
Light + Chlorophyll
CO2 + H2O （ CH2O ） +O2

Purple and green bacteria

Light + bacteriochlorophyll
CO2 + 2H2S （ CH2O ） + H
2O + 2S

Photoheterotroph:
Purple nonsulfur bacteria (Rhodospirillum)
CO2 + 2CH3CHOHCH3 Light + bacteriochlorophyll （ CH2O ）
+ H2O + 2CH3COCH3
 Properties of microbial photosynthetic systems

Property cyanobacteria Green and Purple nonsulfur b

purple bacteria acteria
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 ATP + NADPH ATP ATP
of energy
conversion
 Chemoautotroph:

Bacteria Electron Electron Products

donor acceptor
Alcaligens and Pseudo H2 O2 H2O
monas sp.
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

Nitrifying bacteria

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

 Nutritional types of microorganisms

• Phototrophs: use light as energy source.

• Chenotrophs: obtain energy from the oxidati
on of chemical compounds.
• Lithotrophs: use reduced inorganic substanc
es as their electron source.
• Organotrophs: exteact electrons from organi
c compounds.
Mixotrophic: many purple nonsulfur ba
cteria

1. No oxygen: photoorganotrophic heterot

rophs
2. Normal oxygen: oxidize organic molec
ules and function chemotrophically.
3. Low oxygen: photosynthesis and oxidat
ive metabolism
 Uptake of nutrients

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.
1, Phagocytosis – Protozoa
2, Permeability absorption – Most microorganisms

• passive transport (simple diffusion)

• facilitated diffusion
• active transport
• group translocation
 passive diffusion

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.
 Facilitated diffusion
The rate of diffusion across selectively permeable membranes is
greatly increased by the use of carrier proteins, sometimes called per
meases, which are embedded in the plasina membrane. Since the diff
usion process is aided by a carrier, it is called facilitated diffusion.

The rate of facilitated d

iffusion increases with the co
ncentratioti gradient much m
ore rapidly and at lower conc
entrations of the diffusing mo
lecule than that of passive dif
fusion.
 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.
 A model of 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.

Because there is no energy input,

molecules will continue to enter only
as long as their concentration is
greater on the outside.
 Active transport

Active transport is the transport of solute mol

ecules to higher concentrations, or against a conce
ntration gradient, with the use of metabolic energy
input.

•lower con. higer con.

•Permeases, energy
 Proton gradients
• Symport: linked transport of
two substances in the same d
irection.
• Antiport: linked transport of
two substances in the opposi
te direction.
• Uniport: one substance enter
 Group translocation

A process in which a molecule is transported i

nto the cell while being chemically altered.

The best-known group translocation system is the pho

sphoenolpyruvate: sugar phosphotransferase system (PT
S), which transports a variety of sugars into procaryotic
cells while Simultaneously phosphorylating them using
phosphoenolpyruvate (PEP) as the phosphate donor.
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): c
ytoplasmic, common to all PTSs.
EII (enzyme II) :
EIIa: cytoplasmic and soluble
EIIb: hydrophilic but frequently is attached toEIIc.
EIIc: a hydrophobic protein that is embedded in the
membrane.
Only specific sugars and varies with PTS.
The phosphoenolpyruvate: sugar phosphotransferase syst
em of E. coli.
 Simple comparison of transport systems
Items Passive Facilitated Active Group
diffusion diffusion transport translocation

Non Yes Yes Yes

carrier proteins

transport speed Slow Rapid Rapid Rapid

against gradient Non Non Yes Yes

Specificity Specificity Specificity

transport No specificity
molecules
metabolic No need Need Need Need
energy

Solutes Not changed Changed Changed Changed

molecules
 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 ir
on and supply it to the cell.
• Microorganisms secrete siderophores when little iron is av
ailable in the medium.
iron-siderophore complex bind the receptor of cell surface:
Fe3+ release; complex enter by ABC transporter.
Siderophores (S)
Receptor

Fe 2+/S

Fe 2+/S
 Cuture media
A culture media is a solid or liquid prep
aration used to grow, transport, and store mic
roorganisms.

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 kno
wn.
eg. Medium for E.coli (g/liter)
glucose 1.0; Na2HPO4 16.4; KH2PO4 1.5;
(NH4)2SO4 2.0; MgSO4 · 7H2O 200mg; C
aCl2 10mg; FeSO4 · 7H2O 0.5mg; Final p
H 6.8-7.0
 Complex media
• Media that contain some ingredients of
unknown chemical composition like pe
ptones, meat extract, and yeast extract.
• What is peptones? Protein hydrolysates
eg. LB medium: peptone; yeast extract; s
odium 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-L-galactose, and D-glucurouic ac
id, 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 micr
oorganisms.
“ 加抑” : add componments which inhibite the
growth of unfavored bacteria.
“ 加富” : add components which promote the gr
owth of favored bacteria.
• Differential media: distinguish between different gr
oups of bacteria.
MacConkey agar: lactose, neutral red dye; lactose-f
ermenting 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, nitro
gen, phosphorus and sulfur?
• Nutritional types of microorganisms
• What are growth factors?
• What are facilitated diffusion, Active transport. G
roup translocation, endocytosis and their differen
ce?
• How do cell uptake iron?
• How to prepare a medium for cultivation microor
ganisms?