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Chapter 2: Applied Microbiology

FERMENTATION TECHNOLOGY

Nguyen Van Thanh

Biotechnology R&D Institute (BiRDI)

Can Tho University (CTU)

FERMENTATION TECHNOLOGY

•Aim of chapter: 1*General information about fermentation techniques:(2)

2* Microbial basis of processes (6)

3* Bio-products (14)

4* Type of culture (15)

5* Mode of culture operation (16) (19) (31)

6* Effect of application type on process:

. Substrates (32)

. Inoculum (33)

. Parameters (36)

. Growth patterns, growth kinetics, and modelling

of growth (47) or type of product (45)

• Propose results:Supplying the basic knowledge on application of fermentation:

Research and development on fermentation (48…)

1. Definition of fermentation?

• In history, the knowledge of science is always

upgrade and fullfil during of time

Justus von Liebig (1803-1873)

• Fermentation is only a pure

chemical reaction (chemoreaction)

• Any information about him?

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He is known as the "father of the fertilizer

industry" for his discovery of nitrogen as an

essential plant nutrient, and his formulation of the

Law of the Minimum which described the effect

of individual nutrients on crops. He also

developed a manufacturing process for beef

extract, and founded a company, Liebig Extract

of Meat Company, that later trademarked the

Oxo brand beef bouillon cube

Louis Pasteur (1822-1895)

• Fermentation is a processing,inwhich the organic substrates transform to products by microorganisms

• What field did he explore and become the famous people?

� proved existence of microorganisms (1864)

� Vaccine

�pasteurization

General definition

• Fermentation is the substrate transformation by

living beings (microorganisms) or part of them

with or without oxygen.

≤

2. Microbial basis of processes

• Groups of microorganisms applied for

fermentation:

* bacteria (7)

* yeast (8)

* fillamentous fungi (9)

* actinomycetes (13)

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lactic acid bacteria

• One kind of bacteria that

can produce lactic acid from

carbohydrates, especially

also on cooked starch

• Other functions?

Some examples:

•Lactococcus lactis (cheese)

•Lactobacillus casei (sour milk)

•Pediococcus halophilus (miso, soy-sauce)

•Lactobacillus plantarum (kimchi, sauerkart)

•Oenococcus oeni (vine)

yeasts

• One kind of mold, that can

produce a carbon dioxide

for making bread from

wheat powder adding

sucrose


>

filamentous fungi

• Aspergillus oryzae, one

kind of fungi, that can use

for soy-sauce, miso or

“tuong” making


conidia

Filamentous fungi

• Penicillium chrysogenum,

the fungus was used for

producing penicillin, the first

antibiotic discovered by

Alexander Fleming (11) on

other specie of fungus named

Penicillium notatum

≤

Alexander Fleming memorial (12)

Robert Koch (1843-1910)

A random discovery of Fleming about penicillin in

1928

>

actinomyces

• One kind of the actinomycetes,

that can produce antibiotics

• Other products?

≤

3. Bio-products

• First metabolites: directly produced during or after

fermentation:

– enzymes, biomass, amino acids,

– vitamines for using or further fermentation.

• Secondary metabolites: indirectly produced after

fermentation:

– antibiotic.

4. Type of culture

• Solid-state fermentation (16)

• Liquid-state fermentation (18)

• Solid-state and liquid-state fermentation (17)

liquid–state fermentation

Fermenter

This is a diagram of a simple

fermenter. In industry, these

would be very large and would

have lots of different pipes and

tubes coming out of it for

various functions.

liquid-state fermentation

• Liquid Fermentation is a controlled process which consists of growing cells in a

liquid broth.

• In this tank also known as a bioreactor, as shown in the picture above, sterility of

the material can be maintained and control of temperature, pH, disolved oxygen,

and mixing, is possible.

• This will provide an optimal environment for the specific microorganism to grow

Both filamentous and non filamentous fungi (such as yeast) can grow in liquid

fermentation systems

solid-state fermentation

In contrast to Submerged (liquid state) Fermentation, Solid State Fermentation (SSF) is the

• cultivation of micro organisms under controlled conditions in the absence of free water.

• examples of products of Solid State Fermentation include industrial enzymes, fuels and nutrient

• enriched animal feeds. The application of modern biotechnical knowledge and process control

modern drum solid state fermenters

Box fermentation

>

traditional solid-state fermentation of “tuong”

(mold fermentation) solid substrate cultivation?

<

traditional liquid-state fermentation of “tuong”

>

soy-sauce fermentation

Type of fermentation? >

bag fermentation

Type of fermentation?

mold fermentation in laboratory scale

Box fermentation: Type of fermentation?

Type of fermentation?

modern moisture-aerated solid state fermenter

modern drum solid state fermenter

traditional liquid state fermentation

stirro-aerated liquid state fermentation

simple fermenter

automatic liquid state fermenter (5 liters)

aerating by stirring motor

Larger fermenter (500 litters) What is type of culture? >

5. Mode of fermentation operation

1. Batch fermentation

2. Semi-continuous fermentation

3. Continuous fermentation

4. Aerobic fermentation: the liquid was stirred up or

aerated for supplying oxygen to microorganism

5. Anaerobic fermentation: put (put away) the oxygen

content out of the liquid medium?

substrates

• Local materials (carbon source: natural or industrial carbohydrates (agriculture and industrial wastes ), nitrogen source: … (nitrogen salts), minerals…other stimulated growth substrates…

• Looking for the method of preparation suitable substrates for used microorganisms and type of fermentation

• Pay attention to use cheap materials and give high yield production

inoculum

• Preparation of inoculum: purify the microorganism

strains, fermentation process (34).

• Mode of inoculum: single or mixed inoculum,

immobilized microorganism (35).

Fermentation process

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>

parameters (effecting factors)

• Kind and substrate containing (39)

• Kind of microorganism (38)

• Aeration (40), shaking (42)

• Moisture (41)

• pH

• Temperature (45)

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≤

≤

≤

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Enzyme production depending on the duration

time and temperature

Research and development on fermentation

• Improving and optimizing the fermentation method

(traditional and modern fermentation)

• Improving and selection microorganism strain

• Production suitable inoculum forms for utilization

• Development fermentation process

Surface response diagram of dry biomass depending on (Surface response diagram of dry biomass depending on (NH4)NH4)22SOSO44(X), (X), succrosesuccrose ((Y) ratiosY) ratios at at

0,25%0,25%KHKH22POPO44

Contour diagram of dry biomass depending on (NH4)Contour diagram of dry biomass depending on (NH4)22SOSO44(X(X), ), succrosesuccrose ((Y) ratios atY) ratios at

0,25% 0,25% KHKH22POPO44

(NH4)(NH4)22SOSO44=0,48%=0,48%

SuccroseSuccrose = 11%= 11%

KHKH22POPO44 = 0,25%= 0,25%

Dry biomass=0,85gDry biomass=0,85g

>

Industrializing solid state fermentation?

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Making suitable form for solid state fermentation

Establishing fermenter and suitable fermentative method

Improving microorganism strain

(Cellulomonas)

Production suitable inoculum forms for utilization

<

Development fermentation process

Collection of waste and processing

Increasing application of waste products

LITERATURE

• 1992. Solid substrate cultivation. Doelle H. W.; D.A.

Mitchell and C. E. Rolz eds. Elsevier Applied Science

• 1997. Biochemical engineering and biotechnology

handbook. Vo.1&2

Thank you for your attention !