Lecture 1Prokaryotic (early nucleus) & Eukaryotic (true nucleus) – same metabolic reactions but different structures and complexities Flagella – Varies in number & arrangement (identification), also for motility (true-movement) (NN)Fimbriae (more) or common pilli – adhesion (VF) (NN)Sex pilus – facilitates passage of DNA from one bacterium to another Cell envelope – Glycocalyx: (same composition, different tenacity and thickness)(NN) 1. Slime layer – loose, soluble shield; protects from dehydration, nutrient loss and aids in attachment (VF)2. Capsule – thick, firmly attached; protects from phagocytosis from white blood cells (VF)Cell Wall – Exoskeleton, maintains strength and shape; common component is peptidoglycan (polysaccharide chain of NAG and NAM polymers cross-linked by tetra-peptide bridges between NAM) 1. Gram (differential staining procedure) Positive (purple)a. 90% Peptidoglycan – 40 layers b. 10% Acidic Polysaccharides i. Lipoteichoic Acid – reinforces cross-linking (rigid); allows transfer of ions 2. Gram Negative (pink/red) a. Outer membrane (largest)i. Phospholipid bilayer (Polar head – hydrophilic, Nonpolar tails – hydrophobic)ii. Contains specialized proteins (PP – non-specific channels; LPS – phobic and phillic; LP – acts as anchor to cell wall) b. Periplasmic space i. Hydrolytic enzymes – break down ii. Peptidoglycan – 1 layerc. Cytoplasmic Membrane i. Protects cytoplasm ii. Phospholipid bilayer iii. Contains integral proteins iv. Site of metabolic reactions, synthesis, transport systems, and secretions (VF)d. Cytoplasm i. Internal matrix made of primarily H20 ii. Cell Pool – nutrients/building blocks for metabolism and catabolismiii. Nucleoid – contains genetic info; minimal amount need to surviveiv. Plasmids – extra-chromosomal (NN); can have advantageous traits; transferred (VF)v. Inclusion Bodies/Granules – storage of reserve nutrients in times of abundance; three types:1. Organic (PS) – Glycogen, starch 2. Lipids – PHB3. Inorganic Phosphatesvi. Endospores – dormant structures with no metabolic activity (mechanism of survival – can withstand environmental stressors); bacteria germinates into under adverse conditions (sporogenesis); will germinate back into cell under good conditions; only by Gram +Lab/RecitationShapes of bacteria: Coccus (round), bacillus (rod), spirillum (spiral) Compound microscope – 2 lens system (ocular 10x) * (objective 10x, 45x, 97x – oil, used for looking at bacteria b/c smallest working distance) Normal vs. Transient Flora – what we have on us every day; can pick up or wash off Opportunistic – can infect under opportune conditions (take advantage of host)Thioglycallate Broth – creates an oxygen gradient (Obligate Aerobes – 20%, Microaerophillic – 4%, Obligate Anaerobes, Facultative)Gram Stain Procedure: Crystal Violet (primary stain), Gram’s Iodine (mordant/fixant), Alcohol (de-colorizer), Safarin (counterstain) Flame Streak – diluting technique used to form pure cultures General Purpose Media – used for growth of non-fastidious bacteria States: liquid (turbidity), semi-solid (motility), solid Achieving different atmospheric conditions: Anaerobic Jar w/Gas Pak, Candle Jar (CO2) Lecture 3: Bacterial growth (Lag, Log, Stationary, Death)- Most bacteria grow in artificial cultural media - Elements needed: (Fastidious – more nutritional requirements; Prototrophs – grow in general purpose)o Macronutrients: Needed in large amounts, usually included in GPM, except Co2 and Po4 o Micronutrients: Needed in small amounts, usually included alsoo Vitamins/Growth Factorso Atmospheric Conditions  Obligate Aerobe  Obligate Anaerobe  Micro-aerophillic  Facultative Bacterial Metabolism (all chemical reactions – oxidative reactions – produces energy needed to make organic compounds)- Metabolic Classes: Heterotrophs, pathogenic (organic compounds), Autotrophs (inorganic compounds), Photosynthetic (use own)- Ways to generate energy: Aerobic respiration, anaerobic respiration, fermentation, photosynthesis Aerobic Respiration Anaerobic Respiration Fermentation Photosynthesis Most commonly used mechanism Same, except: Doesn’t produce as much energy, but supports anaerobic growth, good under harsh conditionsPathway Glycolytic Pathway 1. Preparative/investment- Uses 2 ATP’s - Aldolase (6 carbon  2, 3-carbons)- Isomerase: makes same2. Payoff - Gains 4 ATP’s (Net:2) & 2 NADH’s- End: Pyruvate (2) Krebs Cycle (Respiratory Process)1. Pyruvate (2)  (Pyruvate Dehydrogenase Complex)  Acetyl CoA, 2 NADH’s and C02 2. Acetyl CoA (undergoes cycle)- Gains: 6 more NADH’s, 2 ATP’sand 2 FADH’s Electron Transport Chain Phosphorylation- Series of electrons transfers within cytoplasmic membrane- NADH is transferred – gives upelectrons (oxidized) for energy(3); FADH enters later (2)- ADP + Pi  ATP synthase complex  ATPGlycolytic Pathway 1. Preparative/investment- Uses 2 ATP’s - Aldolase (6 carbon  2, 3-carbons)- Isomerase: makes same2. Payoff - Gains 4 ATP’s (Net:2) & 2 NADH’s- End: Pyruvate (2) Krebs Cycle 1. Pyruvate (2)  (Pyruvate Dehydrogenase Complex)  Acetyl CoA, 2 NADH’s and C02 2. Acetyl CoA (undergoes cycle)- Gains: 6 more NADH’s, 2 ATP’sand 2 FADH’s Electron Transport Chain Phosphorylation- Series of electrons transfers within cytoplasmic membrane- NADH is transferred – gives upelectrons (oxidized) for energy(3); FADH enters later (2)- ADP + Phi  ATP synthase complex  ATPGlycolytic Pathway 1. Preparative/investment- Uses 2 ATP’s - Aldolase (6 carbon  2, 3-carbons)- Isomerase: makes same2. Payoff - Gains 4 ATP’s (Net:2) & 2 NADH’s- End: Pyruvate (2) Electron AcceptorOxygen Oxygen-containing salts (inorganic molecules) – sulfate, nitrate, carbonate and ferric ironOrganic compounds LightEnd ProductsC02, H20 Hydrogen sulfide, methane, nitrite Lactic acid, butyric acid, acetic acid, ethanol, butanol(glucose interacts with NADH)ATPEnergy 38 ATP’s Not 38 ATP’s, less 2 ATP’s Organisms Aerobes, facultative Aerobes, facultative Anaerobes, facultative- All use glucose - All conserve energy forming high-energy compounds - All have intermediary steps - PO4 comes from ADP/ATP, recycled by kinases- NADH/NAPDH is producedLecture 3 Mice experiment: genetic information can be transformed!, perfected when given DNAase – DNA contains genetic information- R Strain – lived - S Strain – died - Heat-killed