MIC 303
INDUSTRIAL AND
ENVIRONMENTAL
MICROBIOLOGY
INDUSTRIAL PRODUCTS FROM
MICROBIAL PROCESSES
(CON’T)
Vinegar
an alcoholic liquid that has been allowed to sour.
“Vinegar” = the French word “vin” (wine), “aigre”
(sour).
It is primarily used to flavor and preserve foods and as
an ingredient in salad dressings and marinades.
also used as a cleaning agent.
Two way of production:
i. the microbial fermentation of alcohol
ii. by the dilution of acetic acid
Key equation:
CH3CH2OH + O2 → CH3CO2H + H2O
Types of vinegar
Malt vinegar
Wine vinegar
Other vinegars
 Cider vinegar
Rice vinegar
 Rice vinegar
 Spirit vinegar
Chemical synthesis of vinegar
Balsamic vinegar
Balsamic vinegar
Microbes used
Key organism: Acetobacter (formerly known as
Mycoderma), pertinent strains: Acetobacter aceti,
Acetobacter pastorianus and Acetobacter hansenii
Acetobacters are microscopic bacteria that live on
oxygen bubbles.
Whereas the fermentation of grapes or hops to make
wine or beer occurs in the absence of oxygen, the
process of making vinegars relies on its presence.
Conversion of ethanol to acetic acid is
accompanied by secondary fermentation resulting in
generation
of
aroma-active
compounds
(acetaldehyde, ethyl acetate and other esters and
higher alcohols such as methyl butanol)
Raw materials
Flavour: depends on the source of the alcohol.
Base materials: Herbs and fruit used to flavor
vinegar.
Commonly used herbs include tarragon, garlic,
and basil.
Popular fruits include raspberries, cherries, and
lemons.
Acetozym
In the vinegar factory, this process is induced by
feeding acetozym nutrients into the tanks of
alcohol.
Acetozym nutrients are manmade mother of
vinegar in a powdered form.
It is a natural carbohydrate called cellulose. This
film holds the highest concentration of
acetobacters.
It is skimmed off the top and added to
subsequent batches of alcohol to speed the
formation of vinegar.
The Manufacturing Process
The Generator Method
modern
commercial
production of vinegar.
submerged
fermentation method.
These
methods
are
based on the goal of
infusing as much oxygen
as possible into the
alcohol product.
The Manufacturing Process
The Submerged Fermentation Method
commonly used in the production of wine vinegars.
Production plants are filled with large stainless steel
tanks called acetators.
The acetators are fitted with centrifugal pumps in the
bottom that pump air bubbles into the tank in much
the same way that an aquarium pump does.
As the pump stirs the alcohol, acetozym nutrients are
piped into the tank. The nutrients spur the growth of
acetobacters on the oxygen bubbles.
A heater in the tank keeps the temperature between
80 and 100°F (26-38°C).
The Manufacturing Process
The Orleans Method
Slow process.
manufacture of highquality vinegars.
also called continuous
method.
The Manufacturing Process
Natural Fermentation
Made easily by fermenting
fresh sap into plastic or
earthen jar until it becomes
sour.
Then pack into plastic
bottles and place under
the heat of sun for few
days.
PRODUCTION OF
ANTIBIOTICS PENICILLIN
Antibiotics produced by
Microorganisms
Antibiotic
Cephalosporin
Chloramphenicol
Erythromycin
Griseofulvin
Penicillin
Streptomycin
Tetracycline
Gentamicin
Producing microorganism
Cephalosporium acrimonium
Streptomyces venezuelae
Streptomyces erythreus
Penicillium griseofulvin
Penicillium chrysogenum
Streptomyces griseus
Streptomyces aureofaciens
Micromonospora purpurea
Synthesis Pathway
-ketoglutarate + AcCoA → homocitrate → L-αaminoadipic acid → L-Lysine + β-lactam
The term "penam" is used to describe the core
skeleton of a member of a penicillin antibiotic. This
skeleton has the molecular formula R-C9H11N2O4S,
where R is a variable side chain.
Moa of Penicillin
First antibiotic to have been manufacture in bulk
All penicillin like antibiotics:
 inhibit synthesis of peptidoglycan, an
essential part of the cell wall.
 do not interfere with the synthesis of other
intracellular components.
 do not affect human cells because human
cells do not have cell walls.
Spectrum of Activity
used in the treatment of bacterial infections caused
by susceptible, usually Gram-positive, organisms.
effective against many previously serious diseases
such as syphillis and Staphylococcus infections.
Some members (e.g. amoxicillin) are also effective
against
Gram
negative
bacteria
but
not
Pseudomonas aeruginosa.
Moa of Penicillin
The β-lactam moiety (functional group) of penicillin
binds to the DD-transpeptidase (links the
peptidoglycan molecules in bacteria) → weakens
the cell wall of the bacterium, causes cytolysis or
death due to osmotic pressure.
In addition, the build-up of peptidoglycan
precursors triggers the activation of bacterial cell
wall hydrolases and autolysins, which further digest
the bacteria's existing peptidoglycan.
Gram-positive bacteria are called protoplasts when
they lose their cell wall. Gram-negative bacteria do
not lose their cell wall completely and are called
spheroplasts after treatment with penicillin.
Production of Penicillin
Penicillin was the first important
commercial product produced
by an aerobic, submerged
fermentation and using a fedbatch culture technique.
Used as input material for some
semi synthetic antibiotics.
Secondary
metabolite,
produced only in the stationary
phase when growth of the
fungus is inhibited by stress, not
produced
during
active
growth.
PRODUCTION OF
MONOSODIUM
GLUTAMATE
Monosodium Glutamate (MSG)
MSG has first isolated as glutamic acid in 1866 by
German chemist (Ritthausen) through the acidic
hydrolysis of gliadin, a component of wheat gluten.
In 1908, Kikunae Ikeda, a Japanese chemist
patented a process for isolating MSG from wheat
flour and produced commercially under trade name
“Ajinomoto” (the origin of flavour).
The Manufacturing Process
Preparation and Repulping Processess of Raw
Materials
 Natural crops is used as raw materials (easily
obtained).
 Ex: Tapioca and sago or sugarcane (molasses).
 Tapioca and sago powder are mixed with water
to form starch slurry.
Liquefaction and Saccharification Process
 Starch sluury is converted into glucose solution
through enzymatic action.
Sterilization Process
 The medium will sterilized using steam sterilization
to eliminate contaminants.
The Manufacturing Process (cont)
Fermentation Process
 The heat sterilized raw materials and other
nutrients are put into fermenter.
 Microbes are added to convert glucose into
glutamic acid.
Crystallization Process
 Acidifying of fermentation materials are carried
out to crystallized the glutamic acid produced.
 The glutamic acid crystal cake are separated
from acidified fermentation broth.
Conversion of Glutamic acid to MSG
 By adding glutamic acid cake into NaOH solution
(food grade) to convert glutamic acid into MSG.
The Manufacturing Process (cont)
Cleaning of MSG
 The impurities are removed by using active
carbon to form more clean and clear MSG.
 Active carbon: many micro holes to attach
impurities on its surface.
Crystallization of MSG
 Clean MSG solution is concentrated by heating.
 The crystal of MSG will be formed.
Drying of MSG
 The crystal then are vibrated and transported
through hot air in closed system until dry.
The Manufacturing Process (cont)
Sieving Process
 Seggregating process to separate MSG crystall
accordance to required size.
Packaging
MSG Production Process
Starch (Tapioca Flour, Maize Flour,
Sago Flour)
+ H2O
Starch slurry
Liquefaction (85ºC cooled)
Enzyme saccharification (60 hr, 5560ºC)
Glucose syrup and nutrient addition
(ex Yeast Extract)
Preculture of Brevibacterium
lactofermentum cells
Substrate sterilization (121 ºC)
Inoculation
Batch Fermentation (1-2 weeks)
Filtration (harvest)
Ion Exchangers (to remove
excess mineral salt)
Acidification (pH 2.2 IEP)
Cooling + Crystallization (40
days, 14 ºC)
Glutamate acid slurry
Separation
Crude MSG (Magenta colour)
Decolorization (Activated carbon,
pH 6.4)
Filtration + NaOH
Pure MSG (pH 6.9, 50% MSG)
Final Filtration
Crystallization
Separation
Sedimentation
Drying
Packing
INDUSTRIAL ALCOHOL
FROM WASTES
(OVERVIEW)
INDUSTRIAL ETHANOL
Industrial Production of Ethanol
(Bio Oil Process)
Concentrated sugar cane
molasses + mineral
(Substrate preparation)
Concentrated yeast stream
(Pichina stipitis, S.cerevisiae,
Candida schatae)
Production
fermenter
(anaerobic)
Batch Fermentation
(7 days)
Centrifugation/
Filtration
Ethanol vapourizer
Concentrated
ethanol vapour
Heat exchange
Cooler
Collection tank
Bottling