PERTUMBUHAN

MIKROBA
Pertumbuhan Mikroba
dan Faktor yang Mempengaruhinya
 Microbial growth
❑ Is defined as an increase in the number of cell
in a population
❑ Waktu generasi (generation time):
▪ waktu yang diperlukan oleh mikroba untuk
membelah diri/berganda.
▪ 2→4→8→16 →…..→ 2n
▪ Variatif
▪ Tergantung pada factor nutrisi, genetic,
suhu.
▪ Kisaran menit-hari
▪ E. coli 20 mnt (laboratory culture, under best
nutritional conditions (Brock Biology of Microbiology 14th ed., 2015)

▪ di alam: mungkin lebih lama → why?

The rate of growth of a microbial culture. (a) Data for a population that doubles every 30
min. (b) Data plotted on arithmetic (left ordinate) and logarithmic (right ordinate) scales

The straight-line function reflects the fact

that the cells are growing exponentially and
the population is doubling in a constant time
interval.
 Matematika pertumbuhan mikroba
• Peningkatan jumlah sel dapat diekspresikan secara matematis (based on a geometric
progression of the number 2).
• 2→4→8→16 →…..→ 2n

• N = N02n
• N: final cell number
• No: initial cell number
• n: number of generations during
the period of exponential growth.
• Waktu generasi (g)
• G = t/n
• G: waktu generasi
• T: durasi pertumbuhan
eksponensial (menit, jam, hari)
• n: number of generations during
the period of exponential growth
So….
Berdasarkan data pada slide no.
4, tentukanlah jumlah generasi
dan waktu generasi contoh
tersebut
 Ditanyakan:
1. Jumlah generasi (n)
2. Waktu generasi (g)

N = 4.096 (212)
N0 = 256 (28)
t = 6 – 4 = 2 jam
Jumlah generasi = n
n = 3,3 (log N – log N0)
= 3,3 (log 212 – log 28)
= 3,3 (12 log 2 – 8 log 2) Waktu generasi = G
= 3,3 [(12 x 0,301) – (8 x 0,301)] G = t/n
= 3,3 (3,612 – 2,408) = 2 jam/4
= 3,3 x 1,204 = 0,5 jam
= 3,97 = 30 menit
≈4
When conditions are right bacteria can double in number every 20 minutes
Kurva Pertumbuhan – growing in enclosed vessel (batch culture)

Typical growth curve for a bacterial population. A viable count measures the cells in the culture that are
capable of reproducing. Optical density (turbidity), a quantitative measure of light scattering by a liquid
culture, increases with the increase in cell number.
Pertumbuhan mikroorganisme dalam suatu batch culture
 Fase Lag
• Setelah inokulasi bakteri pada medium fresh media, sel melakukan pertumbuhan.
• Fase lag dapat berlangsung singkat atau lama, bergantung pada histori inoculum serta kondisi
media & pertumbuhan. → exponentially growing culture vs old culture, same medium vs
different, same condition vs different.
• Sel akan beradaptasi pada lingkungannya.
• Semua proses yang dilakukan selama fase lag merupakan persiapan sel untuk membelah diri
• Sel belum membelah, namun sel bertambah besar ukuran dan massanya, melakukan sintesis
enzim, protein, RNA dan meningkatkan aktivitas metabolismenya.
• A lag is also encountered when the inoculum is of low viability (few live cells) or contains cells
that have been damaged but not killed by some stressor, such as high or low temperature,
radiation, or toxic chemicals.
• A lag is also observed when a microbial culture is transferred from a rich culture medium to a
poorer one (a medium downshift); for example, from a complex medium to a defined
medium
 Fase Eksponensial
• Sel mulai membelah dengan
kecepatan konstan dan tergantung
pada komposisi media dan keadaan
pada saat inkubasi.
• Sel akan membelah menjadi 2,4,8,
dan seterusnya megikuti pola
pertumbuahn eksponensial.
• cells are typically in their healthiest
state and are thus most desirable for
studies of their enzymes or other
cell components.
• Kecepatan tergantung pada faktor
lingkungan & genetis.
Fase Stasioner & Kematian
• Pada fase stasioner terjadi
keseimbangan antara jumlah sel yang
mati dan jumlah sel yang hidup.
• Bakteri menghasilkan metabolit
sekunder dan produk yang beracun.
• Sel akan terus menerus memproduksi
metabolit sekunder dan terjadi
akumulasi produk beracun.
• Nutrien pada medium mulai habis,
sehingga terjadi kompetisi antar sel
sehingga beberapa sel mati dan yang
lainnya tetap hidup.
• Nutrien berkurang & penumpukan
metabolit yg bersifat racun → sel mati&
tjd penurunan secara eksponensial
 Continuous Culture
❑ a known volume of fresh medium is added at a constant rate while an equal
volume of spent culture medium (which also contains cells) is removed at the same
rate → attains steady state.
❑ The most common type of continuous culture is the chemostat, a device wherein
both growth rate (how fast the cells divide) and cell density (how many cells per ml
are obtained) can be controlled independently.
❑ 2 faktor yang mempengaruhi growth rate & cell density:
▪ Dilution rate → the rate at which fresh medium is pumped in and spent medium
is removed.
▪ Concentration of a limiting nutrient (misal: sumber C dan N dalam chemostat
vessel.
❑Advantage to the
chemostat is that a cell
population can be
maintained in the
exponential growth phase
for long periods, days or
even weeks.
❑Exponential phase cells are
usually most desirable for
physiological experiments,
and such cells can be
available at any time when
grown in a chemostat.
Suhu
❑ Suhu meningkat → kecepatan reaksi enzimatik meningkat & pertumbuhan makin cepat.
❑ However, saat melewati batas suhu tertentu → protein atau komponen sel lain dapat
terdenaturasi atau rusak.
❑ Cardinal Temperature:
▪ minimum temperature below which growth is not possible
▪ optimum temperature at which growth is most rapid
▪ maximum temperature above which growth is not possible
❑ The cytoplasmic membrane must remain in a semifluid state for nutrient transport and
bioenergetic functions to take place → if an organism’s cytoplasmic membrane stiffens to
the point that it no longer functions properly in transport or can no longer develop or
consume a proton motive force → the organism cannot grow
The cardinal temperatures:
minimum, optimum, and
maximum. The actual values may
vary greatly for different organisms
Temperature classes of organisms

A certain groups are the most commonly studied microbes → which are widespread in nature → which
groups are they?
Psikrofil vs Psikrotoleran
❑ Psikrofil
▪ Suhu optimal: ≤15oC ; Suhu maksimal: < 20oC; Suhu minimal: ≤0oC
▪ produce enzymes that function—often optimally—in the cold and that may be
denatured/inactivated at even very moderate temperatures.
▪ greater content of α-helix and lesser content of β-sheet secondary structure → β -sheet
secondary structures tend to be more rigid than a-helices. the greater α -helix content
allows these proteins greater flexibility for catalyzing their reactions at cold
temperatures.
▪ tend to have greater polar and lesser hydrophobic amino acid content and lower
numbers of weak bonds (i.e. ikatan hydrogen & ion).
▪ Membran sitoplasma: higher content of saturated and shorter-chain fatty acids.
▪ “cold shock” protein → maintaining other proteins in an active form under cold
condition.
▪ Cryoprotectant → antifreeze protein or specific solutes (i.e. glycerol, certain sugar) →
help prevent the formation of ice crystals than can puncture the cytoplasmic
membrane.
Psikrofil vs Psikrotoleran
❑ Psikrotolerant
▪ Tumbuh pada suhu 0oC
▪ Suhu optimal: 20-40oC
▪ are found in environments that are constantly cold and may be killed by warming,
even to as little as 20°C.
▪ More widely distributed in nature.
▪ Kadang perlu inkubasi bbrp pekan hingga membentuk visible growth pada kultur lab.
Snow algae. (a) Snow bank in the Sierra Nevada, California,
with red coloration caused by the presence of snow algae.
Pink snow such as this is common on summer snow banks
at high altitudes throughout the world. (b) Photomicrograph
of red-pigmented spores of the snow alga Chlamydomonas
nivalis. The spores germinate to yield motile green algal
cells. Some strains of snow algae are true psychrophiles but
many are psychrotolerant, growing best at temperatures
above 20°C. From a phylogenetic standpoint, C. nivalis is a
green alga
Growth of hyperthermophiles in boiling water. (a)
Boulder Spring, a small boiling spring in Yellowstone
National Park. This spring is superheated, having a
temperature 1–2°C above the boiling point. The mineral
deposits around the spring consist mainly of silica and
sulfur.
(b) Photomicrograph of a microcolony of prokaryotes that
developed on a microscope slide immersed in such a
boiling spring.
Growth of thermophilic cyanobacteria in a hot spring
in Yellowstone national Park. Characteristic V-shaped
pattern (shown by the dashed white lines) formed by
cyanobacteria at the upper temperature for phototrophic
life, 70–73°C, in the thermal gradient formed from a
boiling hot spring. The pattern develops because the
water cools more rapidly at the edges than in the center of
the channel. The spring flows from the back of the picture
toward the foreground. The light-green color is from a
high-temperature strain of the cyanobacterium
Synechococcus. As water flows down the gradient, the
density of cells increases, less thermophilic strains enter,
and the color becomes more intensely green.
CONTOH MIKROBA DAN APLIKASINYA #SUHU
 Contoh kasus #1:
Efek Variasi Suhu terhadap Kemampuan Degradasi Fenol
Isolat ATA6
Isolat ATA6
12
350
Log jumlah bakteri (CFU/mL)

10 300

Konsentrasi Fenol (ppm)

8 250

200
6 25oC 25oC
30oC 150
4 30oC
35oC 100 35oC
2
50
0
0 24 48 72 96 0
0 24 48 72 96
Waktu (Jam)
Waktu (Jam)

Isolat HP3 Isolat HP3

12 350
Log jumlah bakteri (CFU/mL)

Konsentrasi Fenol (ppm)

10 300

250
8
200
6 25oC 25oC
150
30oC 30oC
4
35oC 100 35oC
2 50

0 0
0 24 48 72 96 0 24 48 72 96
Waktu (Jam) Waktu (Jam)
Hasil degradasi fenol oleh empat isolat
terseleksi setelah inkubasi 96 jam
Konsentrasi fenol (ppm)
Kode Isolat
25oC 30oC 35oC

14,67 8,67
DL120 170

DOK135 103,33 16,67 228,67

ATA6 154,67 48 149,33

HP3 174 41,33 1,33

pH
 Efek Variasi pH terhadap Kemampuan Degradasi Fenol
Isolat ATA6 Isolat ATA6
12 350
Log jumlah bakteri (CFU/mL)
10 300

Konsentrasi Fenol (ppm)

8 250
200
6 pH 5 pH 5
150
pH 7 pH 7
4
100
pH 9 pH 9
2 50
0 0
0 24 48 72 96 0 24 48 72 96
Waktu (Jam) Waktu (Jam)

Isolat HP3
Isolat HP3
12
350
Log jumlah bakteri (CFU/mL)

10 300

Konsentrasi Fenol (ppm)

8 250

pH 5 200
6 pH 5
pH 7 150
4 pH 7
pH 9 100
pH 9
2 50
0 0
0 24 48 72 96 0 24 48 72 96
Waktu (Jam) Waktu (Jam)
Tabel Hasil degradasi fenol oleh empat isolat terseleksi
pada berbagai pH, setelah inkubasi 96 jam

Konsentrasi fenol (ppm)

Kode Isolat
pH 5 pH 7 pH 9

DL120 210 24,67 204,67

DOK135 204 28 168

ATA6 253,33 47,33 211,33

HP3 182 41,33 127,33

 Osmolaritas – Water availability
❑ solvent of life
❑ water availability is an important factor affecting the growth of microorganisms.
❑ Water availability not only depends on how moist or dry an environment is but also a
function of the concentration of solutes (salts, sugars, or other substances) dissolved
in the water that is present.

❑ Water availability → water activity (aw) → the ratio of the vapor pressure of air
in equilibrium with a substance or solution to the vapor pressure of pure water.

❑ When an organism is transferred from a medium of high aw to one of low a , it

maintains positive water balance by increasing its internal solute concentration.
• → by pumping solutes into the cell from the environment

• → or by synthesizing a cytoplasmic solute

❑ m
Effect of NaCl concentration on growth
of microorganisms of different salt
tolerances or requirements. The
optimum NaCl concentration for marine
microorganisms such as Aliivibrio
fischeri is about 3%; for extreme
halophiles, it is between 15 and 30%,
depending on the organism.
Oxygen
Growth versus O concentration. From left to right,
aerobic, anaerobic, facultative, microaerophilic, and
aerotolerant anaerobe growth, as revealed by the position
of microbial colonies (depicted here as black dots) within
tubes of thioglycolate broth culture medium. A small
amount of agar has been added to keep the liquid from
becoming disturbed. The redox dye resazurin, which is
pink when oxidized and colorless when reduced, has been
added as a redox indicator. (a) O 2 penetrates only a short
distance into the tube, so obligate aerobes grow only
close to the surface. (b) Anaerobes, being sensitive to O,
grow only away from the surface. (c) Facultative aerobes
are able to grow in either the presence or the absence of
O2 (a) (b) (c) (d) (e) 2 and thus grow throughout the tube.
However, growth is better near the surface because these
organisms can respire. (d) Microaerophiles grow away
from the most oxic zone. (e) Aerotolerant anaerobes grow
throughout the tube. Growth is not better near the surface
because these organisms can only ferment.
SEKIAN