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Fermentation Technology Chapter I II III IV
Fermentation Technology Chapter I II III IV
623311
Yalun Arifin
Chemical Engineering Dept.
University of Surabaya
Course content
I.
II.
III.
IV.
Introduction
General aspects of fermentation processes
Quantification of microbial rates
Stoichiometry of microbial growth and product
formation
V. Black box growth
VI. Growth and product formation
VII. Heat transfer in fermentation
VIII. Mass transfer in fermentation
IX. Unit operations in fermentation (introduction to
downstream processing)
X. Bioreactor
2
Chapter I
Introduction
What is fermentation?
Pasteurs definition: life without air, anaerobe
red ox reactions in organisms
New definition: a form of metabolism in which the
end products could be further oxidized
For example: a yeast cell obtains 2 molecules of ATP
per molecule of glucose when it ferments it to
ethanol
Organism
Use
Ethanol
Saccharomyces
cerevisiae
Industrial solvents,
beverages
Glycerol
Saccharomyces
cerevisiae
Production of
explosives
Lactic acid
Lactobacillus
bulgaricus
Food and
pharmaceutical
Acetone and
butanol
Clostridium
acetobutylicum
Solvents
-amylase
Bacillus subtilis
Starch hydrolysis
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10
Winemaking fermenter
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Chapter II
General Aspects of Fermentation
Processes
12
Fermenter
The heart of the fermentation process is the fermenter.
In general:
Stirred vessel, H/D 3
Volume 1-1000 m3 (80 % filled)
Biomass up to 100 kg dry weight/m3
Product 10 mg/l 200 g/l
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Types of fermenter
17
Fermentation medium
Define medium nutritional, hormonal, and
substratum requirement of cells
In most cases, the medium is independent of the
bioreactor design and process parameters
The type: complex and synthetic medium (mineral
medium)
Even small modifications in the medium could
change cell line stability, product quality, yield,
operational parameters, and downstream processing.
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Medium composition
Fermentation medium consists of:
Macronutrients (C, H, N, S, P, Mg sources water,
sugars, lipid, amino acids, salt minerals)
Micronutrients (trace elements/ metals, vitamins)
Additional factors: growth factors, attachment
proteins, transport proteins, etc)
For aerobic culture, oxygen is sparged
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Inoculums
Incoculum is the substance/ cell culture that is
introduced to the medium. The cell then grow in the
medium, conducting metabolisms.
Inoculum is prepared for the inoculation before the
fermentation starts.
It needs to be optimized for better performance:
Adaptation in the medium
Mutation (DNA recombinant, radiation, chemical
addition)
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Chapter III
Quantification of Microbial Rates
22
C, N, P, S source
biomass
CO2
O2
product
heat
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kg.i / hour
m3 reactor
kg.i / hour
kg. X
ri = qi CX
Thus:
rX = CX
Product
rP = qPCX
25
rj
q jC X
qj
rate. j
rate.i
ri
qiC X
qi
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Chapter IV
Stoichiometry of Microbial Growth and
Product Formation
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Introduction
Cell growth and product formation are complex processes
reflecting the overall kinetics and stoichiometry of the
thousands of intracellular reactions that can be observed within
a cell.
Thermodynamic limit is important for process optimization.
The complexity of the reactions can be represented by a simple
pseudochemical equation.
Several definitions have to be well understood before studying
this chapter, for example: YSXmax, YATP X, YOX, maintenance
coefficient based on substrate (ms).
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Composition of biomass
Molecules
Protein 30-60 %
Carbohydrate 5-30 %
Lipid 5-10 %
DNA 1 %
RNA 5-15 %
Ash (P, K+, Mg2+, etc)
Elements
C
40-50 %
H
7-10 %
O
20-30 %
N
5-10 %
P
1-3 %
Ash 3-10%
Anabolism
Amino acids protein
Sugars carbohydrate
Fatty acids lipids
Nucleotides DNA, RNA
energy
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Catabolism
Catabolism generates the energy needed for anabolism and
maintenance. It consist of electron donor couple and
electron donor acceptor couple
For example:
Glucose + ()O2 ()HCO3- + H2O
donor couple: glucose/HCO3acceptor couple: O2/H2O
Glucose ()HCO3- + ()ethanol
donor couple: glucose/HCO3acceptor couple: CO2/ethanol
The catabolism produces Gibbs energy (Gcat.reaction)
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Coupled anabolism/catabolism
C-source (anabolism) and electron-donor (catabolism) are
often the same (e.g. organic substrate)
Only a fraction of the substrate ends in biomass as C-source,
while the rest is catabolized as electron-donor to provide
energy for anabolism and maintenance
YSX is the result of anabolic/catabolic coupling.
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34
35
N-source?
YSX =
0.0506 * 88 * 0.95
0.172 C-mol X/mol oxalate
24.6
36
YSX
1
5.815
mol oxalate/C-mol X
0.172
qO2
37
38
+4
+1
-2
-3
+6
Fe
+3
+ charge
-1
- charge
+1
NH4+ as N-source
-3
N2 as N-source
NO3- as N-source
+5
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for compounds
For example: glucose (C6H12O6)
glucose = 6(4) + 12(1) + 6(-2) = 24 = 4/C-glucose
Biomass? O2? Fe2+? Citric acid? Ethanol? Lactic acid?
-balance
It is used to calculate stoichiometry
It follows from conservation relations (C, H, O, N, charge, etc)
by eliminating the unknown stoichiometric coefficient for
reference compounds
It relates biomass, substrate/donor, acceptor, product
(H2O, H+, HCO3-, N-source are always absent)
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Example
Catabolism of glucose to ethanol in anaerobic culture
-C6H12O6 + aC2H6O +bCO2 + cH2O +dH+
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Further reading
Stoichiometry calculations in undefined chemical systems for
fermentation with complex medium, biological waste
water treatment, and soluble and non-soluble compounds
Measurements of lumped quantities:
1. TOC, Carbon balance
2. Kj-N, Kjeldahl-nitrogen for all reduced nitrogen (organic
bound and NH4+), N-balance
3. ThOD, COD balance (similar to balance)
42