IV.

MICROBIAL NUTRITION
A. Nutrient requirement
B. Nutritional types of microorganisms
C. Uptake of nutrients
D. The Study of Microbial Growth

A. 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.

Nutrients: Substances in the environment used by

organisms for catabolism and anabolism.
1. Macronutrients: required in large amounts, including:
carbon, oxygen, hydrogen, nitrogen, sulfur, phosphorus
(Components of carbonhydrates, 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.

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
make up all or part enzyme cofactors, and only
very small amounts are required for growth.

B. NUTRITIONAL TYPES 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

Sulfur-oxdizing bacteria,
(inorganic), Inorganic H/e- Hydrogen bacteria,
donor, CO2 carbon source Nitrifying bacteria

Chemical energy source

Chemoheterotroph
(organic), Organic H/e(Chemoorganotroph) donor, Organic carbon
source

Most bacteria, fungi,

protozoa

Photoautotroph:
Algae, Cyanobacteria

CO2 + H2O

Light + Chlorophyll

CH2O +O2

Purple and green bacteria

CO2 + 2H2S

Light + bacteriochlorophyll

CH2O + H2O + 2S

Photoheterotroph:
Purple nonsulfur bacteria (Rhodospirillum)

CO2 + 2CH3CHOHCH3
+ H2O + 2CH3COCH3

Light + bacteriochlorophyll

CH2O

Properties of microbial photosynthetic systems

Property

cyanobacteria

Green and purple Purple nonsulfur

bacteria
bacteria

Photo - pigment

Chlorophyll

Bacteriochlorophyll Bacteriochlorophyll

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

of energy
conversion

ATP

Chemoautotroph:
Bacteria

Electron
donor

Electron
acceptor

Products

Alcaligens and
Pseudomonas sp.

H2

O2

H2O

Nitrobacter

NO2NH4+
H2
S0. H2S

O2
O2
SO4 2NO3-

NO3- , H2O
NO2- , H2O
H2O. H2S
SO4 2- , N2

Fe2+

O2

Fe3+ , H2O

Nitrosomonas
Desulfovibrio
Thiobacillus denitrificans
Thiobacillus ferrooxidans

Nitrifying bacteria

2 NH4+ + 3 O2

2 NO2- + 2 H2O + 4 H+ + 132 Kcal

C. 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

a. passive transport (simple diffusion)
b. facilitated diffusion
c. active transport
d. group translocation

a. 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.
No carrier protein;

No energy.

b. Facilitated 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.
higher 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.

c. Active transport
Active transport is the transport of solute molecules
to higher concentrations, with the use of metabolic
energy input.
Lower con. higer con.
Permeases, energy

 Symport: linked transport of

two substances in the same
direction.
Antiport: linked transport of
two substances in the
opposite direction.
Uniport: one substance enter

d. Group translocation
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.
PTS: sugar phosphortransferase system
PEP+sugar(outside)pyruvate+sugar-P(inside)

The phosphoenolpyruvate: sugar phosphotransferase

system of E. coli.

Simple comparison of transport systems

Passive
diffusion

Facilitated
diffusion

Active
transport

Group
translocation

carrier
proteins

Non

Yes

Yes

Yes

transport
speed

Slow

Rapid

Rapid

Rapid

transport
molecules

No specificity

Specificity

Specificity

Specificity

metabolic
energy

No need

Need

Need

Need

Solutes
molecules

Not changed

Changed

Changed

Changed

Items

D. THE STUDY OF MICROBIAL GROWTH

Growth takes place on two levels

Cell synthesizes new cell components and
increases in size
The number of cells in the population increases

The Basis of Population Growth: Binary

Fission

Figure 7.13

The Rate of Population Growth

Generation or doubling time: The time required for a
complete fission cycle
Each new fission cycle or generation increases the
population by a factor of 2
As long as the environment is favorable, the doubling
effect continues at a constant rate
The length of the generation time- a measure of the
growth rate of an organism
Average generation time- 30 to 60 minutes under optimum
conditions
Can be as short as 10 to 12 minutes

This growth pattern is termed exponential

Generation Time
Interval waktu yang dibutuhkan mikroba untuk
membelah diri waktu generasi
Rumus :
N = No x 2n
n = 3,3 (log10 N log10No)
g = t/n
R = n/t = 1/g

Keterangan : N
No
n
t
g
R

: populasi akhir
: populasi awal
: pangkat kelipatan
: waktu
: waktu generasi
: laju pertumbuhan

The Population Growth Curve

A population displays a predictable pattern called a
growth curve
The method to observe the population growth pattern:
Place a tiny number of cells in a sterile liquid medium
Incubate this culture over a period of several hours
Sampling the growth at regular intervals during
incubation
Plating each sample onto solid media
Counting the number of colonies present after
incubation

Stages in the Normal Growth Curve

Data from an entire growth period typically
produce a curve with a series of phases:
1. Lag Phase
2. Exponential Growth Phase
3. Stationary Growth Phase
4. Death Phase

1. Lag Phase
Relatively flat period
Newly inoculated cells require a period of
adjustment, enlargement, and synthesis
The cells are not yet multiplying at their
maximum rate

2. Exponential Growth (Logarithmic or log)

Phase
When the growth curve increases
geometrically
Cells reach the maximum rate of cell
division
Will continue as long as cells have adequate
nutrients and the environment is favorable

3. Stationary Growth Phase

The population enters a survival mode in

which cells stop growing or grow slowly
The rate of cell inhibition or death
balances out the rate of multiplication
Depleted nutrients and oxygen
Excretion of organic acids and other
biochemical pollutants into the growth
medium

4. Death Phase
The curve dips downward
Cells begin to die at an exponential rate

Figure 7.15

Potential Importance of the Growth Curve

Implications in microbial control, infection,
food microbiology, and culture technology
Growth patterns in microorganisms can
account for the stages of infection.

Pengukuran Pertumbuhan mikroorganisme

1. Pengukuran jumlah sel
Metode hitung langsung tidak membedakan antara
sel hidup dan mati, dan dapat diselesaikan dengan
observasi mikroskopik langsung pada ruang hitung
Petroff-Hausser atau haemocytometer

Metode langsung lain yaitu dengan turbidimeter atau

spektrofotometer

Turbidimetri
Didasarkan pada difraksi atau
penyebaran cahaya oleh sel-sel bakteri
pada media cair
Cahaya diukur dengan absorbansi pada
spektrofotometer

Penghitungan Langsung dengan Turbidimetri

Penghitungan langsung melalui pengukuran

berat kering massa
Pemisahan sel dari media cair dan
ditimbang (berat basah)
Sel dikeringkan dan ditimbang ~
sebagai berat kering

Pengukuran secara tidak langsung

Menghitung sel hidup antara lain plating sampel secara pour
plate atau spread plate ke dalam media pertumbuhan yang
sesuai dan dilihat formasi koloni. Tipe metode hitung ini
hanya untuk sel yang aktif bereproduksi (hidup)
diekspresikan sebagai colony forming units (CFU).
Metode lain dengan menghitung koloni yang tumbuh pada
membrane filters, atau menggunkan the most probable
number (MPN)

PENGHITUNGAN TIDAK LANGSUNG DENGAN SPREAD-PLATING