IBPT Mod 1
1.Industrial bioprocessing means using living cells (like bacteria, yeast, or biological enzymes) as mini-factories to make useful products at a large commercial scale.
Where It Is Used:
Medicine: Making life-saving drugs like insulin, vaccines, and antibiotics.
Food & Drink: Making bread, cheese, yogurt, and flavorings.
Energy & Environment: Making clean fuels (like bioethanol) and cleaning up wastewater.
Why It Matters:
Eco-Friendly: Works under mild temperatures and normal pressure, using less energy than traditional chemical factories.
High Precision: Biological tools target exact reactions, producing fewer unwanted chemical byproducts.
Renewable: Uses plant waste, sugars, and agricultural leftovers instead of petroleum.
2.Six Major Milestones in Bioprocessing History
Ancient Fermentation: Early humans used wild yeast and bacteria to make bread, beer, and cheese without knowing how the micro-organisms actually worked.
Louis Pasteur's Discovery (1850s): Pasteur proved that living microbes cause fermentation. This turned traditional brewing into a real science.
The ABE Process (1910s): Scientist Chaim Weizmann used a specific bacterium to make industrial solvents (acetone and butanol). This created the need for clean, germ-free industrial equipment.
Penicillin Production (1940s): To mass-produce penicillin during World War II, engineers built deep, bubbling tanks with air pumps. This created the modern bioreactor.
Enzyme Tech (1960s–1970s): Scientists learned to trap enzymes on solid supports so they could be used continuously to make products like high-fructose corn syrup.
Genetic Engineering (1970s–Present): Scientists learned to edit DNA. This allowed us to insert human genes into bacteria to produce complex medicines like human insulin.
3. Major Types of Bioproducts
Antibiotics: Medicines made by microbes to kill harmful bacteria (e.g., Penicillin).
Enzymes: Natural proteins that speed up reactions, used in laundry detergents and food processing (e.g., Amylase)
Amino Acids: Building blocks of protein added to animal feed or food supplements (e.g., MSG, L-Lysine).
Vaccines: Biological preparations that train the immune system to fight diseases (e.g., Hepatitis B vaccine).
Therapeutic Proteins: Lab-made proteins that treat diseases directly (e.g., Recombinant Human Insulin for diabetes).
Biofuels: Renewable fuels made from organic material (e.g., Ethanol made by yeast from sugarcane).
4. Unit Operations: Upstream vs. Downstream
Upstream Operations (Preparing and Growing):
Food Preparation: Mixing nutrients (sugars, salts, water) to feed the cells.
Sterilization: Heating everything to kill unwanted wild germs.
Starter Culture (Inoculum): Growing a small batch of healthy cells in a lab flask, then scaling it up to seed the main tank.
Tank Control: Keeping the main tank stirred, aerated, and at the right temperature and pH.
Downstream Operations (Extracting and Purifying):
Separation: Filtering or spinning (centrifuging) the mixture to separate liquid from solid cells.
Cell Breaking: Pop open the cell walls if the product is trapped inside the cells.
Concentration: Removing water to shrink the volume of liquid.
Purification: Passing the liquid through special filters/columns to isolate the exact target product from everything else.
Drying & Packaging: Drying the final product into a powder or liquid bottle for sale.
5.Primary Metabolites -Primary metabolites are metabolites that has in the size during the log phase or the tropophase of the microbial growth.
They are essential for the growth and metabolism of the corresponding microorganisms
examples includes amino acids,organic acid,ethanol etc
Higer volumetric yields. (Citric acid prodn via Aspergillus Niger ).
Secondary Metabolites -Secondary metabolites are the metabolites that are synthesized during the stationary phase or idiophase of the microbil growth.
they are not essential for the growth of the microorganisms includes antibiotics,anti cancer agents,pigment.
Lower volumetric yields(penicillin prodn via penicillium chrysogenum).
6.Industrial applications of microbial metabolites
Pharmaceutical applications -antibiotics(penicillin), immunosuppressants like Cyclosporin A prevents organ rejection, statins reduce cholestrol.
Food and beverage - organic acids acts as preservatives (citric acid), a.a acts as flavour enhancer, microbial polysaccharide acts as thickening agents.
Agricultural - Microbial metabolites serve as biopesticides, bio-fungicides etc
E. Coli as industrial cost
Gram-negative, facultative anaerobic rod-shaped bacterium.
Rapid growth rate -very short doubing time(20 min)
Low cost processing - simple, cheap, synthetic formulations
well-understood genetics
Limitations-
Lack of post translational modifications- cannot perform mammalian type N and O glycosylation
Endotoxin contamination- outer memb contains LPS which contains toxin
Industrial appl -
Production of non-glycosylated therapeutics like recombinant human insulin, human growth hormone (hGH), and granulocyte colony-stimulating factor (G-CSF).
8.Penicillium and Aspergillus
Penicillium Species:
Primary producer of \beta-lactam antibiotics like Penicillin (P. chrysogenum).
Used in food manufacturing for cheese ripening (P. roqueforti, P. camemberti).
Aspergillus Species:
Major industrial producer of citric acid (A. niger).
Secretes large quantities of enzymes like amylases, pectinases, and cellulases for industrial processing.
8 marks questions.
The Four Phases of Microbial Growth
Lag Phase:
What happens: Cells are adjusting to their new environment. There is no increase in cell number
Biological activity: The cells are actively synthesizing enzymes, RNA, and biological molecules needed to digest the nutrients in the medium.
Exponential (Log) Phase:
What happens: Cells divide at their maximum speed . The population doubles at regular intervals (exponential growth).
Biological activity: Metabolism is balanced and extremely active as cells consume nutrients rapidly to build cell walls, proteins, and DNA.
Stationary Phase:
What happens: Cell division slows down and eventually equals the rate of cell death, making the net growth rate zero.
Biological activity: Triggered by nutrient depletion (e.g., running out of glucose or oxygen) or the buildup of toxic waste products.
Death Phase:
What happens: Nutrients are completely exhausted and toxic waste reaches critical levels.
Biological activity: Cells lose viability and die faster than new ones are produced, leading to a steep decline in biomass.
CHO Cell lines :
CHO cells serve as the primary mammalian expression host, producing approximately 70% of market-approved recombinant therapeutic proteins (e.g., monoclonal antibodies)
Using selection markers like the DHFR/MTX system, CHO genomes are engineered to amplify copy numbers of inserted target genes
They adapt well to large-scale suspension cultures (>10,000 L bioreactors) in fed-batch modes
CHO cells correctly perform glycosylation, which is vital for therapeutic stability, bioactivity, and half-life in the human body.
Complex mammalian proteins often fail to fold correctly in bacteria, forming insoluble aggregates (inclusion bodies). CHO cells provide the proper cellular environment
CHO cells generate glycoforms that closely mimic native human proteins, minimizing immunogenicity risks.