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Bioprocess Engineering

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Presentation on theme: "Bioprocess Engineering"— Presentation transcript:

1 Bioprocess Engineering
Course Outline Basics of Cell Biology and Biochemistry Enzymes Enzyme kinetics Immobilized enzymes Fermentation and Cell Culture Cell Growth ; Batch Growth Kinetics Product formation kinetics; Nutrient limitation Ideal Chemostat; Modeling approaches. Design and Analysis of Bioreactors Suspension bioreactors Immobilized cell bioreactors Scale-Up and Optimization of Bioreactors Oxygen Mass Transfer; Mixing; Sterilization Instrumentation and Control

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Course Outcomes Upon completion of this course, students shall be able to: 1. Describe the growth of microorganisms. 2. Analyze enzyme systems. 3. Determine the reaction stoichiometry for bioreactors and understand the operation of bioreactors. 4. Recognize principles of bioreactor analysis and design. 5. Develop sufficient skills to read current literature in the bioprocess engineering field. 6. Understand where the field is heading and identify future opportunities.

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Required Textbook M. L. Shuler and F. Kargi, Bioprocess Engineering: Basic Concepts, 2e, Prentice-Hall (2002). Course Grading Homework % Quizzes % Class contributions % Mid-term exams % Final Exam %

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Typical Bioprocess – Raw materials preparation – Inoculums preparation – Sterilization – Production in bioreactors – Recovery – Separation – Packaging

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Typical stirred tank bioreactor

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Airlift bioreactor

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Basics of Cell Biology and Biochemistry Eukaryotic and Prokaryotic cells Living cells are divided into two types - prokaryotic and eukaryotic. This division is based on internal complexity. Eukaryotic: The cells of protozoa, higher plants and animals are highly structured. These cells tend to be larger than the cells of bacteria, and have developed specialized packaging and transport mechanisms that may be necessary to support their larger size.

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Basics of Cell Biology and Biochemistry Eukaryotic and Prokaryotic cells Prokaryotic: These cells are simple in structure, with no recognizable organelles. They have an outer cell wall that gives them shape. Just under the rigid cell wall is the more fluid cell membrane. The cytoplasm enclosed within the cell membrane does not exhibit much structure when viewed by electron microscopy. Prokaryotic cells are found in bacteria, including both Archaebacteria and Eubacteria, and including the blue-green algae.

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Basics of Cell Biology and Biochemistry Eukaryotic and Prokaryotic cells Prokaryotic cells are found in bacteria, including both Archaebacteria and Eubacteria, and including the blue-green algae.

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Basics of Cell Biology and Biochemistry Eukaryotic and Prokaryotic cells Prokaryotes share several characteristics:  Prokaryotes DO NOT have a membrane-bound nucleus.  Prokaryotes DO have a cell membrane, but DO NOT have membrane bound organelles.  Prokaryotes HAVE ribosomes that are different from those of Eukaryotes.  Almost all Prokaryotes are smaller than the smallest Eukaryotes.  Most Prokaryotes are single – celled organisms (Unicellular).

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Basics of Cell Biology and Biochemistry Eukaryotic and Prokaryotic cells

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STRUCTURAL CHARACTERISTICS OF A BACTERIAL CELL v v Cell Wall protects the cell and gives shape, made up of polysaccharides

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STRUCTURAL CHARACTERISTICS OF A BACTERIAL CELL Cell Membrane regulates movement of materials into and out of the cell;  contains enzymes important to cellular respiration v v

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STRUCTURAL CHARACTERISTICS OF A BACTERIAL CELL v v Chromosome carries genetic information inherited from past generations

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STRUCTURAL CHARACTERISTICS OF A BACTERIAL CELL v v Plasmid contains some genes obtain through genetic recombination

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STRUCTURAL CHARACTERISTICS OF A BACTERIAL CELL v v Flagellum moves the cell

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STRUCTURAL CHARACTERISTICS OF A BACTERIAL CELL Pilus (Pili) assist the cell in attaching to other surfaces, which is important for genetic recombination v v

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STRUCTURAL CHARACTERISTICS OF A BACTERIAL CELL Cytoplasm contains DNA, ribosomes, and organic compounds required to carry out life processes v v

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STRUCTURAL CHARACTERISTICS OF A BACTERIAL CELL Outer Membrane protects the cell against some antibiotics Capsule, and slime layer protects the cell and assist in attaching the cell to other surfaces Endospore protects the cell against harsh environmental conditions, such as heat or drought

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Basics of Cell Biology and Biochemistry Bacteria (cell) growth Bacteria are all around us. Given good growing conditions, a bacterium grows slightly in size or length, a new cell wall grows through the center forming two daughter cells, each with the same genetic material as the parent cell. If the environment is optimum, the two daughter cells may divide into four in 20 minutes. Scientists who study bacteria try to create the optimum environment in the lab: culture medium with the necessary energy source, nutrients, pH, and temperature, in which bacteria grow predictably.

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Basics of Cell Biology and Biochemistry Bacteria (cell) growth a new cell wall grows through the center forming two daughter cells

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Basics of Cell Biology and Biochemistry Bacteria (cell) growth LAG PHASE: Growth is slow at first, while the cells acclimate to the food and nutrients in their new habitat. LOG PHASE: Once the metabolic machinery is running, they start multiplying exponentially, doubling in number every few minutes. STATIONARY PHASE: As more and more cells are competing for food and nutrients, booming growth stops and the number of bacteria stabilizes. DEATH PHASE: Toxic waste products build up, food is depleted and the cells begin to die.

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Basics of Cell Biology and Biochemistry Bacteria (cell) growth Traditionally, bacteria have been grouped based on their Structure, Physiology, Molecular Composition, and Reaction to specific types of stains, (Gram Stain), rather than on their evolutionary relationships. Bacteria can be one of THREE Different SHAPES:

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Basics of Cell Biology and Biochemistry GRAM STAINING Most species of Eubacteria can be grouped into GRAM-POSITIVE BACTERIA and GRAM-NEGATIVE BACTERIA based on their response to a laboratory technique called GRAM STAINING. Hans Christian Gram, a Danish Microbiologist, developed the Gram-stain technique in 1884.

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Basics of Cell Biology and Biochemistry GRAM STAINING The technique involves STAINING Bacteria with a PURPLE DYE (Crystal Violet), and IODINE, and RINSED with ALCOHOL.  Then Restained with A PINK DYE (Safarinin).

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GRAM STAINING Depending on Structure of their cell walls, the bacteria absorb either the purple dye or the pink dye. Gram-Positive Bacteria will retain the PURPLE DYE and appear Purple. Gram-Negative Bacteria will appear PINK from the PINK DYE.

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Basics of Cell Biology and Biochemistry GRAM-POSITIVE BACTERIA Have a thicker layer of PEPTIDOGLYCAN in their cell walls, made of a protein sugar complex that takes on the purple color during gram staining.

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Basics of Cell Biology and Biochemistry GRAM-POSITIVE BACTERIA Gram-positive bacteria include; Organisms that produce BENEFICAL Substances Example: Bacteria used to make yogurt, pickles and buttermilk. Bacteria used to make ANTIBIOTICS, including Tetracycline and Streptomycin These bacteria are called ACTINOMYCETES. Organisms that cause important human DISEASE, including Scarlet fever, Toxic shock syndrome, and Pneumonia.

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Basics of Cell Biology and Biochemistry GRAM-POSITIVE BACTERIA Many of these bacteria produce TOXINS, which are poisons to our bodies. Toxins can be deadly; a single gram of the toxin produced by Clostridium botulinum  (Botulism) could kill more than one million people. Antibiotics kill other Gram-positive bacteria by preventing them from making proteins.  They affect only the GROWTH of bacteria without harming the body cells of humans.

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ANTIBIOTICS  Antibiotics are drugs that fight bacteria by interfering with their cellular functions. PENICILLIN interferes with Cell Wall Synthesis. TETRACYCLINE interferes with Protein Synthesis. Many Antibiotics are derived from chemicals that bacteria or fungi produce. SULFA DRUGS are Antibiotics that are Synthesized in Laboratories. (Man Made)

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Basics of Cell Biology and Biochemistry GRAM-NEGATIVE BACTERIA Have an extra layer of lipids on the outside of the cell wall and appear pink after gram staining. The extra lipid layer stops the PURPLE Stain from entering the CELL WALL.  They do absorb the PINK Stain, so they are easily distinguished with a microscope.

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Basics of Cell Biology and Biochemistry GRAM-NEGATIVE BACTERIA The extra lipid layer also stops many ANTIBIOTICS from entering the bacteria. Treatment for these requires a different ANTIBIOTIC than those used for infections caused by Gram-positive bacteria. These bacteria photosynthesis differ from plants, they do not release oxygen as a by-product, but produce SULFUR as a by-product. Bacteria that produce sulfur are also called SULFUR PRODUCING BACTERIA.

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Basics of Cell Biology and Biochemistry NUTRITION AND GROWTH TWO major differences between groups of BACTERIA are their SOURCE OF ENERGY AND WHETHER OR NOT THEY USE OXYGEN FOR CELLULAR RESPIRATION. Most BACTERIA are HETEROTROPHS; they get their energy by consuming Organic Matter as a source of Nutrition. Heterotrophic bacteria that feed on dead or decaying material are called SAPROPHYTES. Some are AUTOTROPHS, they obtain their energy by making their own Food from SUNLIGHT OR MINERALS.

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Basics of Cell Biology and Biochemistry NUTRITION AND GROWTH PHOTOAUTOTROPHS ARE BACTERIA THAT USE SUNLIGHT AS AN ENERGY SOURCE. CHEMOAUTOTROPHS OBTAIN THEIR FOOD FROM OXIDIZING INORGANIC COMPOUNDS INSTEAD OF SUNLIGHT.

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Basics of Cell Biology and Biochemistry NUTRITION AND GROWTH The Second important Metabolic difference between BACTERIA groups has to do with CELLULAR RESPIRATION. ORGANISM THAT USE OXYGEN DURING CELLULAR RESPIRATION ARE CALLED AEROBES. THOSE THAT DO NOT USE OXYGEN ARE CALLED ANAEROBES, TYPICALLY THEY GET ENERGY FOR THEIR CELLULAR RESPIRATION THROUGH FERMENTATION.

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Basics of Cell Biology and Biochemistry NUTRITION AND GROWTH THE PRESENCE OR ABSENCE OF OXYGEN IN THE ENVIRONMENT IS A LIFE DETERMINING CONDITION FOR SOME BACTERIA: A. BACTERIA CALLED OBLIGATE AEROBES CANNOT LIVE WITHOUT OXYGEN. B. BACTERIA CALLED OBLIGATE ANAEROBES CANNOT LIVE IN THE PRESENCE OF OXYGEN. C. BACTERIA CALLED FACULTATIVE ANAEROBES CAN USE OXYGEN WHEN IT IS AVAILABLE, BUT THEY DO NOT DEPEND UPON IT.

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Basics of Cell Biology and Biochemistry NUTRITION AND GROWTH Bacteria have varying Temperature Requirements for Growth.  Some bacteria grow best in Cold Temperatures of 0-20 degrees C .  Others grow best in Temperatures between degrees C. Thermophilic (Heat Loving) bacteria grow best in Temperatures between degrees C. Most bacteria Grow BEST at a pH of 6.5 to 7.5 (7.0 is neutral).


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