Mode of Fermentation; Batch, Fed Batch and Continuous - PowerPoint Presentation

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Mode of Fermentation; Batch, Fed Batch and Continuous

Sri Kumalaningsih

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Outline

Overview

Batch fermentationFed batch fermentationContinuous fermentation

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Outline

Overview

Batch fermentationFed Batch cultureContinuous cultureGrowth kinetics

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Batch fermentation

Most fermentations are batch processes

Nutrients and the inoculum are added to the sterile fermenter and left to get on with it!

Anti-foaming agent may be added.Once the desired amount of product is present in the

fermenter

the contents are drained off and the product is extracted

.

After emptying, the

tank is cleaned & prepared

for a new batch.

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Product examples

Food grade ethanol (cosmetics and pharmacy)WineKimchiSaurkrautAstaxantin

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Outline

Overview

Batch fermentationFed Batch cultureContinuous cultureGrowth kinetics

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INDUSTRIAL APPLICATION OF FED-BATCH

Penicillin

Glucose, phenyl acetic acid, ammonia source CephalosporinGlucose, methionine

StreptomycinGlucose, ammonia sourceGlutamic acid

Urea, ethanol, (acetic acid)

Amylase

Carbon source

Bakers Yeast

Glucose

Citric acid

Glucose, ammonia

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Continuous fermentation

Some products are made by a continuous culture system.Sterile medium is added to the fermentation with a balancing withdrawal of broth for product extraction.

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ADVANTAGES / DISADVANTAGES OF CC

Advantages Uniformity of operation Process demands are constant

i.e. continuous cycle of sterilisation, fermentation, harvesting, extraction Once in steady-state demands re process control are constant i.e. oxygen demand

Disadvantages

>>

high cost

Susceptibility to

contamination

Duration of run is longer



increased chance of contamination

Strain degeneration arising from large number of generations

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Overflow

Effluent

Fresh medium from

reservoir

Sterile air

Flow-rate

regulator

Stirrer

Culture

Set up for Continuous culture

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APPLICATION

OF CONTINUOUS CULTURE

INDUSTRY; Waste-treatment Single-cell protein Continuous beer production Continuous amino acids, organic acids production

Continuous ethanol Continuous bakers yeast

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TYPES

OF CONTINUOUS CULTURE

Method of control; Chemostat

- regulated by control of concentration of limiting nutrient

Turbidostat

- regulated by biomass using optical density (photoelectric cell)



Biostat

- regulated by systems monitoring biomass other than optical density (

e.g

CO

2

production)

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Cell

Number

Time in Hours

Steady State

The development of growth in a

chemostat

Inoculation

m

max

Population density increases

Nutrient limitation causes decrease in

m

Growth rate equals loss of cell biomass

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MODIFICATIONS

OF BASIC CHEMOSTAT

MULTI-STAGEDifferent environments or growth rates in the various reactors (e.g. 1st  biomass, 2nd  product)

SINGLE STAGE WITH CELL RECYCLEApplication in activated sludge waste-treatmentRelationship between D and  different when recycle used.

EFFECT OF FEEDBACK;

1. Increase biomass conc. in

fermenter

- lower in effluent

2. Decrease residual substrate

3. Maximise rate of product formation

4.

Dri

f

t

is increased - useful when substrate is dilute

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F

1

S

R

X

1

S

1

V

1

F

O2

S

R2

X

2

S

2

V

2

F

2

Chemostats in series

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Growth Kinetics

Involved by:

Binary fisionGrowth Curve

Spesific growth rate

Substrate utilization

Monod Equation

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20

(1) The steps of binary fision

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21

(2) Growth Curve

Lag phase

Log phase

Stationary phase

Death phase

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22

The steps of growth

2

1

3

4

5

6

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The steps of growth (1)

(1) Fase permulaan

Karakteristik:

Terjadi adaptasi oleh mikroba yang diinokulasikan

Mikroba memproduksi berbagai macam enzim dan zat antara yang berguna bagi pertumbuhan

Sel-sel mikroba mulai membesar

Akan tetapi belum terjadi pembelahan

1

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The steps of growth (2)

(2) Fase pertumbuhan yang dipercepat

Karakteristik:

Bakteri mulai membelah diri

Akan tetapi waktu generasinya masih panjang

Fase permulaan dan pertumbuhan yang dipercepat biasa disebut dengan lag-phase

2

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The steps of growth (3)

(3) Fase pertumbuhan logaritma

Karakteristik:

Pada fase ini kecepatan pembelahan paling tinggi

Waktu generasinya pendek dan konstan

Metabolisme terjadi paling pesat

Keadaan terus berlangsung sampai salah satu nutrien habis

Atau terjadi penimbunan racun akibat metabolisme yang menyebabkan terhambatnya pertumbuhan

3

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The steps of growth (4)

(4) Fase pertumbuhan yang mulai terhambat

Karakteristik:

Kecepatan pembelahan berkurang

Sel yang mati bertambah banyak

Terjadi karena makin berkurangnya nutrien dan bertambahnya racun

Terdapat perubahan kondisi fermantasi (misal perubahan pH, kenaikan suhu)

4

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The steps of growth (5)

(5) Fase stasioner yang maksimum

Karakteristik:

Terjadi penurunan kadar nutrien dan penimbunan zat racun

Kecepatan pembelahan berkurang

Jumlah bakteri yang mati semakin meningkat

Jumlah bakteri yang dihasilkan sama dengan jumlah bakteri yang mati sehingga jumlah bakteri

konstan

5

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The steps of growth (6)

(6) Fase kematian yang dipercepat dan kematian logaritma

Karakteristik:

Disebut juga dengan fase menurun

Kematian terus meningkat

Kecepatan pembelahan menjadi nol

Setelah sampai pada fase kematian logaritma kecepatan kematian mencapai maksimal dan jumlah sel menurun dengan cepat

6

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Steps of growth

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3. Specific growth rate

ln X

Waktu

Slope =



X

Waktu

a

b

c

d

e

f

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2. Specific growth rate



(laju pertumbuhan spesifik pada setiap

waktu

pertumbuhan

dapat

ditentukan

sebagai

berikut

:

Akumulasi

sel

=

pertumbuhan

–

pengeluaran

–

sel

yang

mati

Keterangan

:

F =

laju

alir

V = volume

kultur

 =

laju

kematian

spesifik

Untuk

Kultur

Curah

(

Batch

)

Untuk

Kultur

Fed

Batch

dx/dt=0

μ

= D

di mana D =

F

V

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4. Substrate Utilization

Laju

Penggunaan SubstratAkumulasi Substrat

=

substrat

masuk

–

substrat

yang

dikonsumsi

untuk

pertumbuhan – substrat yang dikonsumsi untuk

sistesis

produk

–

substr

a

t

yang

dikonsumsi

untuk

perawatan

–

substrat

keluar

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4. Substrat Utilization Rate

Laju

Penggunaan Substrat

F =

Laju

alir

(l/jam)

V = Volume

kultur

(l)

S

0

= [

substrat

yang

masuk

] (g/l)

Y

x

/s

=

koefisien

rendemen

biomassa

Y

p

/s

=

koefisien

rendemen

produk

=

laju

pertumbuhan

spesifik

q

p

=

laju

pembentukan

produk

spesifik

m

=

koefisien

pemeliharaan

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4. Specific Utilization Rate

Untuk

Kultur Curah/batch:

Laju

penggunaan

substrat

spesifik

(

q

s

)

Untuk

Kultur

fed

batch:

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Terima Kasih

Thank you