Process Development Scale-up • Laboratory flask: the first indication that a process of commercial interest is possible • Laboratory fermenter: generally of glass.
Download ReportTranscript Process Development Scale-up • Laboratory flask: the first indication that a process of commercial interest is possible • Laboratory fermenter: generally of glass.
Slide 1
Process Development
Slide 2
Slide 3
Slide 4
Scale-up
• Laboratory flask: the first indication that a
process of commercial interest is possible
• Laboratory fermenter: generally of glass and
generally of 1-5-10 liter size / to test variations in
medium, temperature, pH, and so on
inexpensively / examination of feeding strategies
(batch, fed-batch, continuous, etc.) / selection of
production system (stirred tank, airlift, packed bed,
solid state, hollow fiber, etc.)
• Pilot plant: usually 80-300-3000 liter size /
careful instrumentation and computer control is
desirable, so that the conditions most similar to
those in the laboratory fermenter can be obtained
• Commercial fermenter: generally 1,000-10,000500,000 liters
Slide 5
Commercial fermenter
Size of fermenter Product
(1,000 liters)
1-20
Diagnostic enzymes, substances
for molecular biology
40-80
Some enzymes, antibiotics
100-150
200-500
Penicillin, aminoglycoside
antibiotics, proteases, amylases,
steroid transformations, amino
acids
Glutamic acid
Slide 6
Key factors that influence yield during
scale-up process
• Inoculum propagation procedures: quality and
quantity of inoculum
• Choice of medium: cheaper nutrient sources
• Sterilization protocols: degradation of heat-labile
compounds
• Experience in larger fermenters: nutrient,
temperature, pH and oxygen gradients; alter the
generation of foam, shear forces and rate of
removal of carbon dioxide
Slide 7
Microbiological considerations on
process development - 1
• Inoculum build-up: vegetative cells suspension /
heat treatment (boiling for 90s for germination) of
Clostridium sp. spores / non-toxic surface active
agent for fungi or actinomyces spores /
fragmented hyphae (Waring blender) of nonsporulating filaments
• Size of inoculum: 0.5-10% ( up to 20% for
production tank) / fluctuation of environments for
the cells such as change in pH value, increase in
supply of nutrients, decrease of growth inhibitors,
diffusion of essential cofactors out of cells,
enzymes of primary metabolism must be adjusted
to the new conditions / has an influence on the lag
phase
Slide 8
Microbiological considerations on
process development - 2
• Stability of inoculum: log or stationary phase /
supply essential nutrients to auxotroph to prevent
back mutation / supply inducer for enzyme
adaptation / preserve master culture / re-isolate,
purify and test for production
• Contamination: microscopic observation and /or
fermentation appearance, lysogenic phages
(mutations result in decrease of production) and /
or lytic phages (isolation of phage-resistant strains)
Slide 9
Microbiological considerations on
process development - 3
• Medium sterilization: crude, technical-grade
nutrients in production medium / large volume and
longer heating and cooling time / thermal
degradation of essential nutrients and production
of toxic substances / production sterile, not strictly
sterile
• Foam control: antifoams e.g. silicones,
polyoxyalkylene glycol, animal or vegetable oils /
inhibitory effect / additional nutrients
Slide 10
Most important criteria used in scaleup studies on aerobic fermentations
• Constant volumetric oxygen transfer rate or
constant dissolved oxygen level
• Constant power consumption per unit of broth or
constant impeller tip speed
• Fermenter height-diameter aspect ratio？
Slide 11
問題
• 工業生產規模的醱酵槽，其接種量(size of
inoculum)常常達到10~20%。使用高接種量的
優缺點為何？
• 種菌(inoculum)的特性對於主醱酵槽中產物的
生成有哪些影響？
• 下列名詞的內涵：Antiforms, Polyoxyalkylene
glycol, Production sterile
Process Development
Slide 2
Slide 3
Slide 4
Scale-up
• Laboratory flask: the first indication that a
process of commercial interest is possible
• Laboratory fermenter: generally of glass and
generally of 1-5-10 liter size / to test variations in
medium, temperature, pH, and so on
inexpensively / examination of feeding strategies
(batch, fed-batch, continuous, etc.) / selection of
production system (stirred tank, airlift, packed bed,
solid state, hollow fiber, etc.)
• Pilot plant: usually 80-300-3000 liter size /
careful instrumentation and computer control is
desirable, so that the conditions most similar to
those in the laboratory fermenter can be obtained
• Commercial fermenter: generally 1,000-10,000500,000 liters
Slide 5
Commercial fermenter
Size of fermenter Product
(1,000 liters)
1-20
Diagnostic enzymes, substances
for molecular biology
40-80
Some enzymes, antibiotics
100-150
200-500
Penicillin, aminoglycoside
antibiotics, proteases, amylases,
steroid transformations, amino
acids
Glutamic acid
Slide 6
Key factors that influence yield during
scale-up process
• Inoculum propagation procedures: quality and
quantity of inoculum
• Choice of medium: cheaper nutrient sources
• Sterilization protocols: degradation of heat-labile
compounds
• Experience in larger fermenters: nutrient,
temperature, pH and oxygen gradients; alter the
generation of foam, shear forces and rate of
removal of carbon dioxide
Slide 7
Microbiological considerations on
process development - 1
• Inoculum build-up: vegetative cells suspension /
heat treatment (boiling for 90s for germination) of
Clostridium sp. spores / non-toxic surface active
agent for fungi or actinomyces spores /
fragmented hyphae (Waring blender) of nonsporulating filaments
• Size of inoculum: 0.5-10% ( up to 20% for
production tank) / fluctuation of environments for
the cells such as change in pH value, increase in
supply of nutrients, decrease of growth inhibitors,
diffusion of essential cofactors out of cells,
enzymes of primary metabolism must be adjusted
to the new conditions / has an influence on the lag
phase
Slide 8
Microbiological considerations on
process development - 2
• Stability of inoculum: log or stationary phase /
supply essential nutrients to auxotroph to prevent
back mutation / supply inducer for enzyme
adaptation / preserve master culture / re-isolate,
purify and test for production
• Contamination: microscopic observation and /or
fermentation appearance, lysogenic phages
(mutations result in decrease of production) and /
or lytic phages (isolation of phage-resistant strains)
Slide 9
Microbiological considerations on
process development - 3
• Medium sterilization: crude, technical-grade
nutrients in production medium / large volume and
longer heating and cooling time / thermal
degradation of essential nutrients and production
of toxic substances / production sterile, not strictly
sterile
• Foam control: antifoams e.g. silicones,
polyoxyalkylene glycol, animal or vegetable oils /
inhibitory effect / additional nutrients
Slide 10
Most important criteria used in scaleup studies on aerobic fermentations
• Constant volumetric oxygen transfer rate or
constant dissolved oxygen level
• Constant power consumption per unit of broth or
constant impeller tip speed
• Fermenter height-diameter aspect ratio？
Slide 11
問題
• 工業生產規模的醱酵槽，其接種量(size of
inoculum)常常達到10~20%。使用高接種量的
優缺點為何？
• 種菌(inoculum)的特性對於主醱酵槽中產物的
生成有哪些影響？
• 下列名詞的內涵：Antiforms, Polyoxyalkylene
glycol, Production sterile