Professional Documents
Culture Documents
Lecture1 Fermenter Design
Lecture1 Fermenter Design
Lecture1 Fermenter Design
Introduction
Pharmaceutical proteins produced via fermentation in transgenic microbes or mammalian cell culture systems provide economical systems for production of therapeutic proteins. These include antibodies, vaccines, blood proteins, etc. Biopharmaceuticals are medical drugs (see pharmacology) produced using biotechnology. They are proteins (including antibodies), nucleic acids (DNA, RNA or antisense oligonucleotides) used for therapeutic or in vivo diagnostic purposes, and are produced by means other than direct extraction from a native (non-engineered) biological source Dozens of new pharmaceuticals produced via fermentation in transgenic microbes have been approved for therapeutic use in the USA. Hundreds of additional biotech drug candidates are in various stages of research or clinical trials. Fermentation systems can be scaled up to produce quantities of pharmaceuticals that are difficult or impossible to produce via traditional methods. Pharmaceutical quality may also be improved. For example, pharmaceuticals produced from blood must be carefully purified to ensure no transmission of viruses as accidental contaminants in the pharmaceutical product. Microbial systems that do not allow human viruses to replicate enable pharmaceutical production with little or no risk of virus contamination.
Introduction
Fermentation technology is the oldest of all biotechnological processes. The term is derived from the Latin verb fevere, to boil--the appearance of fruit extracts or malted grain acted upon by yeast, during the production of alcohol. Fermentation is a process of chemical change caused by organisms or their products, usually producing effervescence and heat. Microbiologists consider fermentation as 'any process for the production of a product by means of mass culture of micro-organisms'. Biochemists consider fermentation as 'an energy-generating process in which organic compounds act both as electron donors and acceptors'; hence fermentation is an anaerobic process where energy is produced without the participation of oxygen or other inorganic electron acceptors.
Metabolite
Ethanol - Saccharomyces cerevisiae alcoholic beverages - Kluyveromyces fragilis Citric acid - Aspergillus niger food industry Acetone and Clostridium butanol acetobutyricum solvents Lysine Corynebacterium nutritional additive Glutamic acid glutamacium flavour enhancer Riboflavin Ashbya gossipii nutritional Eremothecium ashbyi Vitamin B12 Pseudomonas denitrificans nutritional Propionibacterium shermanii Dextran Leuconostoc mesenteroides industrial Xanthan gum Xanthomonas campestris industrial
Secondary metabolites
Penicillin Penicillium chrysogenum antibiotic Erythromycin Streptomyces erythreus antibiotic Streptomycin Streptomyces griseus antibiotic Cephalosporin Cephalosporium acrimonium antibiotic Griseofulvin Penicillium griseofulvin antifungal antibiotic Cyclosporin A Tolypocladium inflatum immunosuppressant Gibberellin Gibberella fujikuroi plant growth regulator Secondary metabolism may be repressed in certain cases. Glucose represses the production of actinomycin, penicillin, neomycin and streptomycin; phosphate represses streptomycin and tetracyclin production. Hence, the culture medium for secondary metabolite production should be carefully chosen.
PRODUCTION ENZYMES
Aspergillus oryzae Amylases Aspergillus niger Glucamylase Trichoderma reesii Cellulase Saccharomyces cerevisiea Invertase Kluyveromyces fragilis Lactase Saccharomycopsis lipolytica Lipase Aspergillus species -Pectinases and proteases Bacillus species Proteases Mucor pusillus Microbial rennet Mucor meihei Microbial rennet
RECOMBINANT PRODUCTS
Therapeutics Proteins mAbs Enzymes Hormones DNA for gene therapy
BIOTRANSFORMATION
Production of a structurally similar compound from a particular one, during the fermentation process is transformation, or biotransformation, or bioconversion. The oldest instance of this process is the production of acetic acid from ethanol. Immobilised plant cells may be used for biotransformation. Using alginate as the immobilising polymer, digitoxin from Digitalis lanata was converted into digoxin, which is a therapeutic agent in great demand. Similarly, codeinone was converted into codeine and tyrosine from Mucuna pruriens was converted into DOPA.
Vitamins Figure 11.13, Vitamin B12 Used as supplements for human food and animal feeds Nearly $1B/year Synthesized chemically, but some by biocatalysis Selected high-yield strains for B12 produce up to 60 mg/L For riboflavin, up to 7 g/L
Amino Acids L-Glutamate (MSG) flavor enhancer Aspartame (phe + asp) sweetener L-Lysine nutritive additive DL- Methionine nutritive additive
Design of Fermenter
Design of Fermenter
Factors to consider when designing a fermenter Aseptic and regulatory capability, longterm reliability Adequate aeration and agitation Low power consumption Temperature and pH controls Sampling facilities
Economy of scale
Scale Up
Definition Adaptation of biological methods of production to large-scale industrial use Objectives Obtain the best biological catalyst Create the best possible environment Purify the products in the most economical ways
Typical Bioprocessing
Penicillin Production
Cost Estimations
Cellular metabolism of substrate Extracellular production of insulin Air sparging for oxygen delivery Impellers for mixing of nutrients and oxygen
Bioreactor Specifications
Di
1.3 m
HL*= m
5.5 Dt
1.50 m
HL**= 6.06m
Novonordisk Bioreactor
Steps
1. Organism selection, with regard to:
substrate versatility byproduct formation characteristics robustness of the organism, e.g., to process upsets viability with regard to cell recycling physiological characteristics (maximum growth rate, aeration requirements, etc.) genetic accessibility
2. Metabolic and cellular engineering: improve existing properties of the organism introduce novel functions, for example, by simplify- ing product recovery, expanding substrate and product ranges, and enabling fermentation to occur under nonstandard conditions
3. Fermentation process development: culture and media optimization (from complex to defined minimal media) optimization of cultivation parameters that take into account product recovery and purification (minimize byproduct formation, minimize chemical inputs, and develop high-cell-density cultivation) incorporation of cell retention/recycling
Types of Fermenter
1. 2. 3. 4. 5. 6. 7. 8. 9. Activated sludge Fermenter Air Lift Fermenter Bubble cap Fermenter Loop Fermenter Mist Fermenter Packed Bed Fermenter Rotating Drug Fermenter Tower Fermenter Trickling Film Fermenter
Classification of Fermenters
2. Air-lift Fermenters
2. Air-lift Fermenters
4. Loop Fermenter
5. Mist Fermenter
8. Tower/Column Fermenters
References
Stanbury, P.F., A. Whitaker, and S. J. Hall, Principles of Fermentation Technology, 2nd ed., Butterworth Heinemann, Oxford, 2000.
References
References
References