Leather Biotechnology

Leather Biotechnology

• Why this course????

 Leather Biotechnology
Fermentation Process: Fermentation is a biochemical process where
microorganisms convert organic compounds into simpler
substances, such as acids, gases, or alcohol. This process is
used in various industries including food production,
pharmaceuticals, and biofuels.
 Bacterial Growth Phases
• Microbial growth in a controlled fermentation environment
typically progresses through four distinct phases:
• 1. Lag Phase: Bacteria adapt to new environment,
preparing for growth.
• 2. Log Phase: Rapid cell division and growth at
maximum rate.
• 3. Stationary Phase: Nutrient depletion slows
growth; secondary metabolite production.
• 4. Death Phase: Nutrients exhausted; cell population
declines.
Leather Biotechnology
 Lag Phase
• When bacteria are first introduced into a new
medium, they enter the lag phase, where they do not
immediately begin dividing.
• This period is characterized by cellular adaptation as
bacteria prepare for growth.
• During this phase, cells synthesize necessary enzymes,
co-factors, and macromolecules required for cell
division and energy production.
• The lag phase duration depends on various factors,
including the bacteria’s previous growth conditions,
the composition of the new medium, and the
physiological state of the cells.
• A shorter lag phase is desirable in industrial
fermentation because it maximizes the active growth
phase, improving the efficiency of the entire process.
 Log (Exponential)
• During the exponential or log phase, bacteria
undergo rapid cell division, growing at a constant and
maximum rate. In this phase, the bacterial population
doubles at a regular interval known as the “doubling
time” or “generation time.”
• Nutrient availability is high, and waste products are
minimal, allowing cells to grow optimally. Bacteria
display high metabolic activity, which can be harnessed
for the production of certain metabolites.
• The log phase is crucial for industrial fermentation
processes that require high biomass or growth-associated
products like enzymes and primary metabolites. Most
commercial fermentations aim to prolong the log phase
to maximize yield.
 Stationary Phase
• The stationary phase occurs when the rate of cell
division equals the rate of cell death. This happens as
nutrients are depleted and waste products
accumulate, creating a less favorable environment
for growth.
• Metabolic activity shifts, and cells begin
producing secondary metabolites, compounds that
are not directly involved in growth but can have
industrial significance, such as antibiotics, pigments,
and certain organic acids.
• Many secondary metabolites, which are
commercially valuable, are produced primarily in the
stationary phase.
 Death Phase
• As the environment becomes increasingly
inhospitable, due to nutrient exhaustion and
waste accumulation, cell death exceeds cell
division, leading to a decline in the bacterial
population.
• Cells lose viability, and the concentration of
viable organisms decreases over time.
• In a closed fermentation system, the death
phase typically marks the end of productivity.
By understanding the death phase dynamics,
scientists and engineers can determine the
optimal time to harvest microbial products to
maximize yields and efficiency
 Fermentation and fermentation products
Five major groups:

1. Production of microbial cells or

biomass.
(baker’s yeast)

2. Production of microbial enzymes.

(amylase, protease, pectinase)

3. Production of microbial metabolites

(primary : amino acids; secondary:
antibiotics)

4. Production of microbial recombinant

products
(insulin, human serum albumin)

5. Biotransformation of a products
(vinegar)
 Biomass

• Baker’s yeast
• Seed culture
• SCP & MBP
Enzymes
 Amylase
 Glucanase
 Protease
 Lactases
 Pectinase
 Invertase
 Cellulase
 Lipase
 Microbial Metabolites

• Ethanol
• Citric acid
• Glutamic acid
• Lysine
• Nucleotides
• Vitamins
 Microbial Recombinant Products

• Insulin
• Interferon
• Humane serum albumin
• Epidermal growth factor
• Calf chymosin
 Biotransformation Products
• Steroids
• Antibiotics
• Prostaglandins
• Vinegar
 Fermentation Process

Fermentation process consists of two parts.

1. Upstream processing
The processes in which biological
materials inoculated and growth in
culture under controlled conditions to
manufacture certain types of products.
2. Downstream processing
The process in which the products are
harvested, tested, purified and packaged,
and also include waste disposal or
recycle.
 Components of a Fermentation Process

Six basic components of a fermentation process:

Upstream processing
1. The formulation of suitable culture media for inoculum preparation
and fermentation.

2. The sterilization of the media, fermenters and ancillary

equipment.

3. The production of culture insufficient quantity to inoculate the

production medium in the fermenter.

4. The growth of organism in the production fermenter under

optimal conditions for product formation.
Downstream processing
5. The extraction of the product and its purification.

6. The disposal of effluents by the process.

Schematic Presentation of a Typical Fermentation Process

1. Medium Formulation

4. Fermentation
Process
2. Sterilization
5. Product recovery
& purification

Culture fluid

Stock Shake Seed Cell separation Biomass

culture flask fermenter
3. Inoculum development Cell-free 6. Effluent
supernatant treatment

Product Product
purification packaging

Upstream Processing Downstream Processing

Aerobic vs. Anaerobic Fermentation

Aerobic Fermentation
• Requires oxygen to proceed and is conducted
by microorganisms that use oxygen for
growth and energy production.
• Aerobic fermentation is common in
processes that require high energy, such as
the production of biomass and specific
growth-associated products like vinegar,
citric acid, and some antibiotics.
• Industrial setups use bioreactors equipped
with aeration systems to maintain optimal
oxygen levels.
• Acetic acid production from ethanol by
Acetobacter bacteria is an aerobic
fermentation process used in vinegar
production.
Aerobic vs. Anaerobic Fermentation

Anaerobic Fermentation
• Occurs in the absence of oxygen and
is used by organisms that either do
not require oxygen (obligate
anaerobes) or can tolerate low
oxygen levels (facultative anaerobes).
• Anaerobic fermentation is crucial in
producing ethanol, lactic acid,
biogas, and certain pharmaceuticals.
• It is common in the food industry for
products like yogurt, sauerkraut,
and alcoholic beverages.
Batch culture
• It is a closed culture system which contain an initial,
limited amount of nutrient.
• The culture is passed through a number of growth
phases.
• Production rate is relatively low. (antibiotic, enzymes)
Continuous culture

• It is a steady state
culture system which
consists of continuous
supply of nutrients &
continuous withdrawal
of products.

• The culture is passed a

long log growth phase.

• Production rate is
relatively very high.
(vinegar, O. acids)
Semi-batch culture
• It is a culture system in which the culture is harvested at
regular intervals and replaced by an equal volume of fresh
medium.
• The culture is passed a relatively moderate log growth
phase.
• Production rate is relatively higher than that of batch
culture. (vinegar, O. acids)
Feed-batch culture
• The batch culture which are feed continuously or
sequentially, with medium without the removal of
culture fluid so that the volume is increased.
Submerged Fermentation
vs
SSF
Submerged Fermentation (SmF)
• Microorganisms grow in a liquid medium with nutrients
dissolved or suspended. This method is well-suited for
bacteria and yeast, which thrive in aqueous
environments with consistent access to nutrients.
• Widely used in the production of antibiotics, ethanol,
and industrial enzymes.
• Ethanol production using yeast in liquid glucose medium
is a classic example of submerged fermentation.
Solid State Fermentation (SSF)
• Microorganisms grow on a solid substrate with minimal
water content, typically on organic waste or other
agricultural residues. This technique is ideal for fungi,
which thrive in low-water conditions.
• Commonly used for enzyme production, bioactive
compounds, and bioconversion of agricultural waste.
• Fungal production of enzymes like cellulase and
amylase using solid substrates such as rice bran or
wheat straw.
 Different Types of Fermenters
Based on size
 Laboratory & research (bench scale) 1-50 L
 Pilot plant fermenter (pilot scale) 50- 1000 L
 Industrial production (Production scale) >1000L
Based on growth system Based on aeration
 Surface fermenter (supported growth system)  Aerobic
Tray fermenter
Film fermenter  Anarobic
 Submerged fermenter (suspended growth system)
Fermenter with mechanically moved internal members
Fermenter with forced convection of liquid (by pump)
Fermenter with pneumatic operation (by compression of air)
Based on fermenter design
Stirred tank with impeller Plunging jet fermenter
Stirred tank with draught tube Nozzle loop reactor
High power Stirred tank Paddle-wheel reactor
Gas-aspiring tube reactor Immerged-slot reactor
Stirred multistage fermenter Enzyme=membrane reactor
Tube loop fermenter Immerged coil reactor
Jet loop fermenter Packed-bed column
Air-lift loop fermenter Trickling film column reactor
Bubble –column fermenter Immersing-surface fermenter
Sieve plate tower fermenter Surface culture fermenter
Important Requirements for Fermenter
Optimal conditions for fermentation requires efficient transfer of mass,
heat and momentum from one phase to the other phase (liquid, gas
and solid).

A fermentor (bioreactor) should provide:

(i) Agitation (for mixing of cells and medium)
(ii) Aeration (aerobic fermentors) for oxygen supply
(iii) Regulation of factors (temp, pH, pressure, aeration, nutrient
feeding, liquid level, etc.)
(iv) Sterilization and maintenance of sterility, and
(v) Withdrawal of cells/medium (for continuous fermentors)
(vi) Computer linkage for efficient process monitoring, data acquisition,
etc.
 Component Parts of a Typical Fermenter
• Power motor
• Vessel
• Impeller
• Water jacket
• Steam inlet
• Nutrient inlet
• Air inlet
• Effluent outlet
• Product outlet
• Temp detector
• pH detector
• Foam detector
• DO meter
• Rotameter
• Sample collection
Purposes of the Parts of Fermenter/Bioreactor
 Service Provisions Required for a Fermentation Plant
• Compressed air
Sterile compressed air
• Chilled water (12-15 C)
Cold water (4 C)
Hot water
Steam (high pressure)
Steam condensate
• Buffering facility
• Electricity
Standby generator
Motors
• Drainage of effluents
• Storage facility for medium
components
• Control & monitoring equipment
• Extraction & recovery equipment
• Accessibility for delivery of
materials
• Maintenance facilities
 Monitoring and Controlling System of Fermentation

• Temperature
• pH
• DO
• Agitation
• Foam
• Pressure
• Biomass
• Air flow
• Weight
• Redox
Process Sensors and Their Possible Control Functions
A sensor is an electronic component, module, or subsystem whose purpose is
to detect events or changes in its environment (physical and chemical) and
send the information to other electronics, frequently a computer process.
(sense, indicate, record and control?)

Physical process sensors Chemical process sensors

 Temperature  pH
(heat/cool control) (acid or alkali addition)
 Pressure  Redox
(inlet/exit gases)
 Agitation shaft power (additives to change redox
(rpm control) potential)
 Foam probe  Oxygen
(foam control) (changes feed rate)
 Weight  Exit gas analysis
(change flow rate) (changes feed rate)
 Flow rate  Medium analysis
(change flow rate) (change in medium composition)
• Light
(change biomass)
Monitoring and Control of Temperature, pH & DO
1. Principles of Fermentation Technology by Peter F. Stanbury and
Allan Whitaker
2. Microbiology: An Introduction - G.J. Tortora, B.R. Funke, and
C.L. Case; Pearson, Boston