Culture Medium

(Substrate)

Industrial
Microbiology

Minggu ke 4
 Introduction

– Medium for laboratory scale application is normally

expensive
– Medium for an industrial scale application should be
cost effective so the production cost can be reduced
– Generally a common medium for industrial scale is in a
liquid form because it is easy to handle
– Medium must contain all essential elements which are
required for microbial growth and production.
Komposisi Media
Element Physiological functions Dry
weight
(%)
H To form organic molecules and water 8
O To form organic molecules and water 20
C To form organic molecules 50
N To form protein, nucleic acids and co-enzyme 14
S To form protein and some co-enzyme 1
P To form nucleic acid, phospolipid dan co-enzyme 3
Mg Co-factor some enzyme reaction (ATP) 0,5
Mn Co-factor of some enzyme 0,1
Ca Co-factor of some enzyme 0,5
Fe Co-factor of some enzyme
 Carbon sources
– Microbes in industries require organic
compound as a carbon source and energy,
except algae and autothropic bacteria
– Carbon accounts for almost 50% as the
highest component required, so it has to be
considered when formulating medium
– Biomass total (X) is directly corellated with
the concentration of carbon source: : X = Ys
Note: Y is a yield constant for substrate and organism
Y value in some medium
Medium Culture cell/ml Dry weight Y
(g/l) (%)
Nutrien Broth Escherichia 5 x 109 1 12,5
8 g/l dry weight coli

Medium sintetik Serratia 2 x 1011 29 37,5

dg 57 g/l glucose marcescens
+ 20 g/l acetate

Mineral solutions Candida 45 45

+ 100 g/l glucose utilis
 Carbon source
– Microorganism are able to use several
carbon source from the simplest one like C2
component (acetate, ethanol) to the
complex one (polysaccaride), ex:
Pseudomonas sp are able to degrade
detergent
– Other microorganism needs one spesific
source for their metabolism, ex:
Methylomonas dan Methylococcus needs
methane and methanol as their carbon and
energy source
 Energy source

– Two pathways for energy (ATP) production:

– Substrate fosforilation: substrate oxidation, 1
molecule glucose produce 2 ATP molecule via
glycolisis reaction
– Oxidative fosforilation: aerobic respiration
– 1 molecule glucose produce 36 ATP molecule
aerobically
 How to determine carbon source?

– First excess substrate are used, cell (dry

weight) obtained 40g/L
– Biomass are assumed to contain 50%
carbon, organism able to convert 50%
substrate to carbon biomass
anaerobically
– How much glucose are actually needed?
 Calculation
– 40 X (100/50) x (50/100) = 40 g C/l
– Glucose: (40 X 180)/72 = 100 g/l
 Molasses
– Sugar industry product that can be used
as substrate for several process, ex:
alcohol, amino acid, etc

– Contain C: sucrose, glucose, fructose

– Contain N: amino acid, ex glutamate,
asparagine, aspartat, alanine, glycine
– Vitamins: biotin, pyridoxine, pantothenic
acid, thiamine
– Contain S
 Other carbon source
– Whey: cheese industry byproduct, contain lactose,
protein, fat and minerals
– Starch: Endomycopsis fibulliger hydrolyze potatoes starch
– Crude oil:10 – 25% C10 – C18
– Hydrocarbons CnH2n+2: slowly degraded, production of
single cell protein ex: Pseudomonas, Bacillus,
Acinetobacter,Corynebacterium, Mycobacterium,
Nocardia dsb
– Hydrocarbon fermentation requires more oxygen
compare to carbohydrate fermentation
– Glucose
C6H12O6 + 2,1O2 + NH3  1,2C5H9NO4 + 5,4 H2O
Molecule result is120% or 98% weight
– Hexadene
C16H34 + 8,45O2 + NH3  3,2C5H9NO4 + 4,1 H2O
Molecule result is 320 % or 208% weight but require 4
times more oxygen

Example: Glutamic acid production

Using hexadene require 3x more oxygen compare to
glucose and produce 3,5x more heat
 Nitrogen source
– Microorganism are able to use anorganic
nitrogen, or assimilate ammonia
– Lactic acid bacteria needs specific amino
acid to grow but it cannot be produced by
bacteria itself.
– Nitrogen source in fermentation: Soy flour,
fish flour, tofu dregs
– Streptococcus faecalis needs specific amino
acid arginin in their medium to grow
 Growth Factor
– Growth factor is needed but microorganism can not
synthesize itself
– Bacteria can be grown in ammonium salt and
glucose if growth factor such as Vitamin, amino acid
or fatty acid are added
– Example: Lactobacillus arabinosa needs really low
concentration of biotin 10-7 g/l.
– Some growth factors are already contained in the
medium formulation, but sometimes it needs to be
added to the medium, ex from yeast extract, liver
extract, etc.
Excessive amount of growth factor
inhibit fermentation product
 Minerals

– It’s not easy to calculate minerals needed in medium

but it will affect the amount of product that can be
produced.
– The approach is carried out by measuring microbial
elements contents (5% biomass weight) and
generally consists of 60% phosphor, 20% sulfur and
metals
– Trial and error experiments with syntetic medium
should be done
– Phosphor and sulfur normally needed in high 0,5 g/l
– Aspergillus niger produce citric acid in Fe deficient
medium
Thank you

Industrial
Microbilogy
 Quiz (50 mins)
1. Please explain conditions where SSF
maybe appropriate for! (point 20)
2. What are the construction materials for
fermenters? Please also explain about the
function of each material? (point 20)
3. What is the difference between mycelium
(bibit) and spawn? (point 10)
4. What are the difference between mushroom
cultivation and processing industries in
Indonesia and Europe? (point 15)
5. Name 5 bacteria and their roles in bio-
industry! (point 15)
6. What elements serve as macronutrients?
What are their functions? (point 20)