Ph and Enzymes | Quizalize

Lide 1: Introduction to Bioreactor A bioreactor is a vessel used for growing microorganisms, plant or animal cells Provides controlled conditions for biological reactions Maintains optimum pH, temperature, oxygen, and nutrients Widely used in fermentation, enzyme, vaccine, and antibiotic production Ensures sterile and aseptic environment Scale ranges from laboratory to industrial production Slide 2: Basic Design Requirements of a Bioreactor Must be constructed with non-toxic, corrosion-resistant materials Should allow effective mixing and mass transfer Provision for sterilization (in situ sterilization) Must maintain uniform temperature and pH Easy sampling without contamination Should support scalability and automation Slide 3: Materials Used in Bioreactor Construction Stainless steel (SS-316) for industrial bioreactors Glass for laboratory-scale bioreactors Plastic (polycarbonate) for disposable bioreactors Materials must withstand heat and pressure Should be smooth to prevent microbial attachment Resistant to chemicals and cleaning agents Slide 4: Main Parts of a Bioreactor Vessel: holds the culture medium and microorganisms Agitator (impeller): provides mixing Sparger: supplies sterile air Baffles: prevent vortex formation Sensors: monitor pH, temperature, dissolved oxygen Ports: used for inoculation, sampling, and feeding Slide 5: Agitation System Ensures uniform mixing of nutrients and cells Improves oxygen transfer rate Common impellers: Rushton turbine, marine propeller Speed controlled by motor Prevents settling of cells Affects shear stress on cells Slide 6: Aeration System Supplies oxygen for aerobic fermentation Air introduced through sparger Types of spargers: ring, nozzle, sintered Maintains dissolved oxygen concentration Air is filtered for sterility Essential for high cell density cultures Slide 7: Temperature and pH Control Temperature controlled by heating/cooling jackets pH maintained using acid or alkali addition Sensors continuously monitor parameters Automated control systems used Ensures optimal microbial growth Prevents enzyme denaturation Slide 8: Foam Control System Foam formed due to protein and agitation Excess foam reduces oxygen transfer Mechanical foam breakers used Chemical antifoam agents added Foam sensor detects foam formation Maintains efficient fermentation Slide 9: Types of Bioreactors – Based on Mode of Operation Batch bioreactor Fed-batch bioreactor Continuous bioreactor Choice depends on product type Widely used in industrial fermentation Controls productivity and yield Slide 10: Batch Bioreactor All nutrients added at the beginning No addition or removal during process Simple and easy to operate Low risk of contamination Used for antibiotics and enzymes Limited control over nutrient depletion Slide 11: Fed-Batch Bioreactor Nutrients added during fermentation Prevents substrate inhibition High product yield Widely used in industrial fermentation Allows better control of growth rate Used in insulin and enzyme production Slide 12: Continuous Bioreactor Fresh medium continuously added Culture removed at same rate Maintains steady-state conditions High productivity Risk of contamination is high Used in wastewater treatment and SCP production Slide 13: Types of Bioreactors – Based on Design Stirred tank bioreactor Airlift bioreactor Bubble column bioreactor Packed bed bioreactor Fluidized bed bioreactor Photobioreactor Slide 14: Stirred Tank Bioreactor (STR) Most commonly used bioreactor Mechanical agitation using impellers Suitable for aerobic fermentation Excellent mixing and oxygen transfer Used for bacteria and fungi Easy scale-up Slide 15: Airlift Bioreactor Mixing achieved by air circulation No mechanical agitator Low shear stress Energy efficient Suitable for shear-sensitive cells Used in wastewater treatment Slide 16: Bubble Column Bioreactor Air bubbles provide mixing Simple design and low cost No moving parts Limited mixing efficiency Used for microbial fermentation Suitable for large-scale operations Slide 17: Packed Bed Bioreactor Contains immobilized cells or enzymes Substrate flows through packed matrix High cell density Used in continuous processes Limited oxygen transfer Used in enzyme and wastewater treatment Slide 18: Fluidized Bed Bioreactor Immobilized particles kept in suspension Better mass transfer than packed bed Reduced clogging Suitable for continuous operation Used in biotransformations Higher operational complexity Slide 19: Photobioreactor Designed for photosynthetic organisms Provides light source Used for algae and cyanobacteria Controls light, CO₂, and temperature Used in biofuel and pigment production Can be tubular or flat-plate design Slide 20: Applications of Bioreactors Production of antibiotics and vaccines Enzyme and organic acid production Single cell protein production Wastewater treatment Biofertilizer and biopesticide production Biopharmaceutical manufacturing

Life Processes Identify and define the seven life processes (MRS GREN). Classification Group living organisms based on observed similarities and differences. Classify vertebrates into taxonomic groups based on visible physical characteristics. Construct a dichotomous key to classify vertebrates. Cells Compare the structure of generalised plant and animal cells, and selected microbes (e.g. bacteria, fungi and Amoeba) Distinguish among cell wall, cell membrane, nucleus, cytoplasm, temporary and permanent vacuoles, mitochondrion, chloroplast, endoplasmic reticulum and ribosomes. Relate the structure of organelles to their functions; Identify specialised cells such as blood cells, ciliated epithelial cells, nerve cells, root hair cells, sperm cells and egg cells. Explain the importance of cell specialisation in multicellular organisms; include hierarchy of cells, tissues, organs; organ systems and then organism Diffusion, Osmosis, Active transport and Osmoregulation Explain the processes of diffusion, osmosis and active transport. Identify everyday instances of these processes occurring. Discuss the importance of diffusion, osmosis and active transport in living systems. Nutrition in Plants Describe the process of photosynthesis in green plants; test for end products, starch or reducing sugar (glucose). Relate the structure of the leaf of a flowering plant to its function in photosynthesis; draw and label the external features and the internal structure (cross section) of a leaf as seen in cross section under the light microscope. Nutrition in Humans Discuss the importance of a balanced diet in humans. State components of a balanced diet (carbohydrates, fats, proteins, vitamins and minerals, water and roughage and their roles) along with the results of their deficiency or surplus. Suggest dietary recommendations for treating and preventing named deficiency and physiological diseases (such as those outlined in the manual and your notes). Perform tests to distinguish among food substances - Test for proteins (Biuret), fats (grease spot), starch (iodine), reducing sugars (Benedict’s solution). The Digestive System in Humans Relate the structures of the human alimentary canal to their functions; Draw and label simple diagrams of the alimentary canal and internal structure of a tooth required. Describe mastication and the role of teeth in the mechanical breakdown of food to be included. (Compare types of teeth in humans and compare types of teeth in herbivores and carnivores.) Explain the role and importance of enzymes role of digestive enzymes in the mouth, stomach and pancreatic enzymes in the small intestine. Discuss properties of enzymes. Deduce from tables and graphs the effects of temperature and pH on enzyme activity. Experimental Skills Follow all drawing rules as outlined in the drawing skills checklist posted in the classroom (including calculation of magnification).

Candidates should be able to: (a) state the main roles of carbohydrates, fats and proteins in living organisms: • carbohydrates as an immediate source of energy • fats for insulation and long-term storage of energy • proteins for growth and repair of cells (b) describe and carry out tests for: • starch (using iodine in potassium iodide solution) • reducing sugars (using Benedict’s solution) • protein (using biuret solution) • fats (using ethanol) (c) state that large molecules are synthesised from smaller basic units: • cellulose, glycogen and starch from glucose • polypeptides and proteins from amino acids • lipids such as fats from glycerol and fatty acids (d) explain the mode of action of enzymes in terms of an active site, enzyme-substrate complex and enzyme specificity using the ‘lock and key’ hypothesis (e) investigate and explain the effects of temperature and pH on the rate of enzyme catalysed reactions

CARBOHYDRATES Carbohydrates are organic compounds composed of carbon, hydrogen, and oxygen in a ratio of about one carbon atom to two hydrogen atoms to one oxygen atom. The number of carbon atoms in a carbohydrate varies. Some carbohydrates serve as a source of energy. Other carbohydrates are used as structural materials. Carbohydrates can exist as monosaccharides, disaccharides, or polysaccharides. Monosaccharides A monomer of a carbohydrate is called a monosaccharide (MAHN-oh-SAK-uh-RIED). A monosaccharide—or simple sugar— contains carbon, hydrogen, and oxygen in a ratio of 1:2:1. The gen- eral formula for a monosaccharide is written as (CH2O)n, where n is any whole number from 3 to 8. For example, a six-carbon mono- saccharide, (CH2O)6, would have the formula C6H12O6. The most common monosaccharides are glucose, fructose, and galactose, as shown in Figure 3-6. Glucose is a main source of energy for cells. Fructose is found in fruits and is the sweetest of the monosaccharides. Galactose is found in milk. Notice in Figure 3-6 that glucose, fructose, and galactose have the same molecular formula, C6H12O6, but differing structures. The different structures determine the slightly different properties of the three compounds. Compounds like these sugars, with a single chemical formula but different structural forms, are called isomers (IE-soh-muhrz). SECTION 2 OBJECTIVES ● Distinguish between monosaccharides, disaccharides, and polysaccharides. ● Explain the relationship between amino acids and protein structure. ● Describe the induced fit model of enzyme action. ● Compare the structure and function of each of the different types of lipids. ● Compare the nucleic acids DNA and RNA. VOCABULARY carbohydrate monosaccharide disaccharide polysaccharide protein amino acid peptide bond polypeptide enzyme substrate active site lipid fatty acid phospholipid wax steroid nucleic acid deoxyribonucleic acid (DNA) ribonucleic acid (RNA) nucleotide C HO H C H OH C OH H C CH2OH H C H OH O Glucose C OH C O H OH C OH H CH2OH C H CH2OH Fructose C H HO C OH H C OH H C CH2OH H C H OH O Galactose Glucose, fructose, and galactose have the same chemical formula, but their structural differences result in different properties among the three compounds. FIGURE 3-6 Copyright © by Holt, Rinehart and Winston. All rights reserved. 56 CHAPTER 3 Disaccharides and Polysaccharides In living things, two monosaccharides can combine in a condensa- tion reaction to form a double sugar, or disaccharide (die-SAK-e-RIED). For example in Figure 3-4, the monosaccharides fructose and glu- cose can combine to form the disaccharide sucrose. A polysaccharide is a complex molecule composed of three or more monosaccharides. Animals store glucose in the form of the polysaccharide glycogen. Glycogen consists of hundreds of glucose molecules strung together in a highly branched chain. Much of the glucose that comes from food is ultimately stored in your liver and muscles as glycogen and is ready to be used for quick energy. Plants store glucose molecules in the form of the polysaccha- ride starch. Starch molecules have two basic forms—highly branched chains that are similar to glycogen and long, coiled, unbranched chains. Plants also make a large polysaccharide called cellulose. Cellulose, which gives strength and rigidity to plant cells, makes up about 50 percent of wood. In a single cellu- lose molecule, thousands of glucose monomers are linked in long, straight chains. These chains tend to form hydrogen bonds with each other. The resulting structure is strong and can be broken down by hydrolysis only under certain conditions. PROTEINS Proteins are organic compounds composed mainly of carbon, hydrogen, oxygen, and nitrogen. Like most of the other biological macromolecules, proteins are formed from the linkage of monomers called amino acids. Hair and horns, as shown in Figure 3-7a, are made mostly of proteins, as are skin, muscles and many biological catalysts (enzymes). Amino Acids There are 20 different amino acids, and all share a basic structure. As Figure 3-7b shows, each amino acid contains a central carbon atom covalently bonded to four other atoms or functional groups. A single hydrogen atom, highlighted in blue in the illustration, bonds at one site. A carboxyl group, —COOH, highlighted in green, bonds at a second site. An amino group, —NH2, highlighted in yel- low, bonds at a third site. A side chain called the R group, high- lighted in red, bonds at the fourth site. The main difference among the different amino acids is in their R groups. The R group can be complex or it can be simple, such as the CH3 group shown in the amino acid alanine in Figure 3-7b. The differences among the amino acid R groups gives different proteins very different shapes. The different shapes allow pro- teins to carry out many different activities in living things. Amino acids are commonly shown in a simplified way such as balls, as shown in Figure 3-7c. (a) Many structures, such as hair and horns are made of proteins. (b) Proteins are made up of amino acids. Amino acids differ only in the type of R group (shown in red) they carry. Polar R groups can dissolve in water, but nonpolar R groups cannot. (c) Amino acids have complex structures, so, in this and other textbooks, they are often simplified into balls. FIGURE 3-7 (b) Alanine (an amino acid) (c) Simplified version of amino acid CH3 H N OH C C H O H (a) Copyright © by Holt, Rinehart and Winston. All rights reserved. BIOCHEMISTRY 57 H H N C C OH H O H CH3 H2O Glycine Alanine H N OH C C H O H H H N C C H O H CH3 N OH C C H O H (a) (b) (a) The peptide bond (shaded blue) that binds amino acids together to form a polypeptide results from a condensation reaction that produces water. (b) Poly- peptides are commonly shown as a string of balls in this textbook and elsewhere. Each ball represents an amino acid. FIGURE 3-8 Substrate Products Enzyme 1 2 3 In the induced fit model of enzyme action, the enzyme can attach only to a substrate (reactant) with a specific shape. The enzyme then changes and reduces the activation energy of the reaction so reactants can become products. The enzyme is unchanged and is available to be used again. 3 2 1 FIGURE 3-9 Dipeptides and Polypeptides Figure 3-8a shows how two amino acids bond to form a dipeptide (die-PEP-TIED). In this condensation reaction, the two amino acids form a covalent bond, called a peptide bond (shaded in blue in Figure 3-8a) and release a water molecule. Amino acids often form very long chains called polypeptides (PAHL-i-PEP-TIEDZ). Proteins are composed of one or more polypep- tides. Some proteins are very large molecules, containing hun- dreds of amino acids. Often, these long proteins are bent and folded upon themselves as a result of interactions—such as hydrogen bonding—between individual amino acids. Protein shape can also be influenced by conditions such as temperature and the type of solvent in which a protein is dissolved. For exam- ple, cooking an egg changes the shape of proteins in the egg white. The firm, opaque result is very different from the initial clear, runny material. Enzymes Enzymes—RNA or protein molecules that act as biological catalysts—are essential for the functioning of any cell. Many enzymes are proteins. Figure 3-9 shows an induced fit model of enzyme action. Enzyme reactions depend on a physical fit between the enzyme molecule and its specific substrate, the reactant being catalyzed. Notice that the enzyme has folds, or an active site, with a shape that allows the substrate to fit into the active site. An enzyme acts only on a specific substrate because only that substrate fits into its active site. The linkage of the enzyme and substrate causes a slight change in the enzyme’s shape. The change in the enzyme’s shape weakens some chemical bonds in the substrate, which is one way that enzymes reduce activation energy, the energy needed to start the reaction. After the reaction, the enzyme releases the products. Like any catalyst, the enzyme itself is unchanged, so it can be used many times. An enzyme may not work if its environment is changed. For example, change in temperature or pH can cause a change in the shape of the enzyme or the substrate. If such a change happens, the reaction that the enzyme would have catalyzed cannot occur.

A solution is a mixture in which one or more substances are uniformly distributed in another substance. Solutions can be mixtures of liquids, solids, or gases. For example, plasma, the liquid part of blood, is a very complex solution. It is composed of many types of ions and large molecules, as well as gases, that are dissolved in water. A solute (SAHL-YOOT) is a substance dissolved in the solvent. The particles that compose a solute may be ions, atoms, or molecules. The solvent is the substance in which the solute is dissolved. For example, when sugar, a solute, and water, a solvent, are mixed, a solution of sugar water results. Though the sugar dissolves in the water, neither the sugar molecules nor the water molecules are altered chemically. If the water is boiled away, the sugar molecules remain and are unchanged. Solutions can be composed of various proportions of a given solute in a given solvent. Thus, solutions can vary in concentra- tion. The concentration of a solution is the amount of solute dis- solved in a fixed amount of the solution. For example, a 2 percent saltwater solution contains 2 g of salt dissolved in enough water to make 100 mL of solution. The more solute dissolved, the greater is the concentration of the solution. A saturated solution is one in which no more solute can dissolve. Aqueous (AY-kwee-uhs) solutions—solutions in which water is the solvent—are universally important to living things. Marine microorganisms spend their lives immersed in the sea, an aqueous solution. Most nutrients that plants need are in aqueous solutions in moist soil. Body cells exist in an aqueous solution of intercellu- lar fluid and are themselves filled with fluid; in fact, most chemical reactions that occur in the body occur in aqueous solutions. Copyright © by Holt, Rinehart and Winston. All rights reserved. Liquid water Solid water Ice (solid water) is less dense than liquid water because of the structure of ice crystals. The water molecules in ice are bonded to each other in a way that creates large amounts of open space between the molecules, relative to liquid water. FIGURE 2-12 solvent from the Latin solvere, meaning “to loosen” Word Roots and Origins CHEMISTRY OF LIFE 43 ACIDS AND BASES One of the most important aspects of a living system is the degree of its acidity or alkalinity. What do we mean when we use the terms acid and base? Ionization of Water As water molecules move about, they bump into one another. Some of these collisions are strong enough to result in a chemical change: one water molecule loses a proton (a hydrogen nucleus), and the other gains this proton. This reaction really occurs in two steps. First, one molecule of water pulls apart another water molecule, or dissociates, into two ions of opposite charge: H2O ∏ H OH The OH ion is known as the hydroxide ion. The free H ion can react with another water molecule, as shown in the equation below. H H2O ∏ H3O The H3O ion is known as the hydronium ion. Acidity or alkalin- ity is a measure of the relative amounts of hydronium ions and hydroxide ions dissolved in a solution. If the number of hydronium ions in a solution equals the number of hydroxide ions, the solution is said to be neutral. Pure water contains equal numbers of hydro- nium ions and hydroxide ions and is therefore a neutral solution. Acids If the number of hydronium ions in a solution is greater than the number of hydroxide ions, the solution is an acid. For example, when hydrogen chloride gas, HCl, is dissolved in water, its mol- ecules dissociate to form hydrogen ions, H, and chloride ions, Cl, as is shown in the equation below. HCl ∏ H Cl These free hydrogen ions combine with water molecules to form hydronium ions, H3O. This aqueous solution contains many more hydronium ions than it does hydroxide ions, making it an acidic solution. Acids tend to have a sour taste; how- ever, never taste a substance to test it for acidity. In concentrated forms, they are highly corrosive to some materials, as you can see in Figure 2-13. Bases If sodium hydroxide, NaOH, a solid, is dissolved in water, it dissociates to form sodium ions, Na, and hydroxide ions, OH, as shown in the equation below. NaOH ∏ Na OH Copyright © by Holt, Rinehart and Winston. All rights reserved. Eco Connection onnection Acid Precipitation Acid precipitation, more commonly called acid rain, describes rain, snow, sleet, or fog that contains high levels of sulfuric and nitric acids. These acids form when sulfur dioxide gas, SO2, and nitrogen oxide gas, NO, react with water in the atmosphere to produce sulfuric acid, H2SO4, and nitric acid, HNO3. Acid precipitation makes soil and bodies of water, such as lakes, more acidic than normal. These high acid levels can harm plant and animal life directly. A high level of acid in a lake may kill mollusks, fish, and amphibians. Even in a lake that does not have a very elevated level of acid, acid precipitation may leach aluminum and magnesium from soils, poisoning water- dwelling species. Reducing fossil-fuel consump- tion, such as occurs in gasoline engines and coal-burning power plants, should reduce high acid levels in precipitation. Sulfur dioxide, SO2, which is produced when fossil fuels are burned, reacts with water in the atmosphere to produce acid precipitation. Acid precipitation, or acid rain, can make lakes and rivers too acidic to support life and can even corrode stone, such as the face of this statue. FIGURE 2-13 44 CHAPTER 2 This solution then contains more hydroxide ions than hydronium ions and is therefore defined as a base. The adjective alkaline refers to bases. Bases have a bitter taste; however, never taste a substance to test for alkalinity. They tend to feel slippery because the OH ions react with the oil on our skin to form a soap. In fact, commercial soap is the product of a reaction between a base and a fat. pH Scientists have developed a scale for comparing the relative con- centrations of hydronium ions and hydroxide ions in a solution. This scale is called the pH scale, and it ranges from 0 to 14, as shown in Figure 2-14. A solution with a pH of 0 is very acidic, a solution with a pH of 7 is neutral, and a solution with a pH of 14 is very basic. A solution’s pH is measured on a logarithmic scale. That is, the change of one pH unit reflects a 10-fold change in the acidity or alkalinity. For example, urine has 10 times the H3O ions at a pH of 6 than water does at a pH of 7. Vinegar, has 1,000 times more H3O ions at a pH of 3 than urine at a pH of 6, and 10,000 times more H3O ions than water at a pH of 7. The pH of a solution can be measured with litmus paper or with some other chemical indicator that changes color at various pH levels. Buffers The control of pH is important for living systems. Enzymes can function only within a very narrow pH range. The control of pH in organisms is often accomplished with buffers. Buffers are chemi- cal substances that neutralize small amounts of either an acid or a base added to a solution. As Figure 2-14 shows, the composition of your internal environment—in terms of acidity and alkalinity— varies greatly. Some of your body fluids, such as stomach acid and urine, are acidic. Others, such as intestinal fluid and blood, are

pH and buffer

PH and Indicators

pH and POH (1)

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