BACTERIAL GROWTH & METABOLISM

GROWTH
GROWTH CURVE

Growth is both at cellular & population level Lag: cells accommodate to new environment Log (exponential) populations double every generation Stationary: growth rate = death rate Death (decline): death rate > growth rate; number of viable cells decreases Generation time: time required for a cell to grow & divide (via binary fission) Dependent on chemical & physical properties Leads to exponential growth; number of cells doubles each generation n Nn = Ni x 2 ; n = generations, Ni = initial number of cells; Nn = number of cells at generation n Steps of binary fission Cell replicates its chromosome, both chromosomes are attached to cytoplasmic membrane Cell elongates, pushing the chromosomes apart Cell forms new cytoplasmic membrane & wall (septum) across midline If daughter cells remain attached: dividing in parallel planes forms a chain, dividing in different planes forms a cluster
ENUMERATION

Measuring microbial reproduction Indirect method: turbidity Direct method: serial dilution & viable plate counts Serial dilution & viable plate count Serial dilution: stepwise dilution of culture; dilution factor = amount of culture added to tube / total volume (1ml sample + 9ml broth = 1:10 dilution, 1:100 dilution, 1:1000 dilution, etc) Viable plate count: live cells will grow on plate; colonies counted (plates w/ 25-250 colonies) to estimate number of cells in original sample CFU/ml (colony forming unit = viable cell) # colonies on plate x 1/diluation factor -4 4 5 65 colonies on plate with 10 overall dilution factor (1/1000 x 1/10) = 65 x 10 = 6.5 x 10 CFU/ml
NUTRITION (C, N, O, ETC)

Carbon source Heterotrophs: gain carbon from existing organic molecules (proteins, carbs, amino/fatty acids) Autotrophs: gain carbon from CO2 (inorganic carbon) Energy source Chemotrophs: gain energy from chemical bonds in molecules Phototrophs: gain energy from sunlight
ATMOSPHERE

Aerobes: utilize oxygen & can detoxify oxygen radicals Aerobic respiration 2 Catalase: enzyme that detoxifies peroxide anion (O2 ) Superoxide dismutase: enzyme that detoxifies superoxide radicals (O2 ) Obligate aerobes: requires oxygen as final electron acceptor of electron transport chain Microaerophiles: require oxygen levels of 2-10% (found in stomach) Anaerobes: dont require or utilize oxygen; or are incapable of detoxifying radicals Anaerobic respiration Fermentation Obligate anaerobe: cant tolerate oxygen; uses something else as final electron acceptor Facultative anaerobe: can live with or without oxygen Aerotolerant anaerobes: prefers anaerobic conditions but can tolerate exposure to low levels of oxygen
TEMPERATURE

Effect of temperature on: proteins (denature); lipid-containing membranes of cells/organelles: rigid/fragile when low; too fluid when high Psychrophiles: -5 to 15 C Mesophiles: 20 to 45 C Thermophiles: 50 to 75 C Hyperthermophiles: 80 to 120 C

WATER ACTIVITY (AW)

Organisms need available (free) liquid water to carry out metabolic pathways Bacteria: 0.95 or greater Fungi: 0.8 or greater Most organisms inhibited at 0.5 or below Reference items: pure water 1, tap water 0.99, honey/dried fruit 0.5 0.7
PH

Neutrophils Acidophiles Alkalinophiles

SALINITY
Halophiles: require saline environment (greater than 9% NaCl)

OTHER
Capnophiles: grow best with high levels of CO2 and low levels of O Barophiles: require extreme hydrostatic pressure found deep below water Extremophiles: require extreme conditions of temperature, pH, and/or salinity
METABOLISM
ENZYMES

Enzymes: protein catalysts that lower activation energy of reaction; mediate all chemical reactions Induced fit model: after binding to substrate, enzyme slightly changes shape so as to bind more tightly Endoenzymes: active inside cell; found within cell cytosol Exoenzyme: active outside cell; secreted by cell into environment 6 main classes (reflecting their mode of action) Hydrolases (lipase): catabolize by adding water via hydrolysis Lyases (decarboxylase): split large molecules w/o using water (catabolic) Isomerases (topoisomerase): rearrange atoms in molecule (neither catabolic or anabolic) Ligases or polymerases (DNA ligase): join 2 molecules together (anabolic); use ATP energy Oxidoreductases (alcohol dehydrogenase): remove electrons (oxidize) or add electrons to (reduce) various substrates (catabolic & anabolic) Transferases (phosphorylase): transfer functional group (amino acid/phosphate/acetyl) between molecules (can be anabolic) Structure: many enzymes are complete in themselves but others are composed of protein & nonprotein portions Apoenzyme: protein portion Cofactors: inorganic helper; metal ions common (irons, magnesium, zinc, copper) Coenzymes: organic helper; NAD/FAD Haloenzyme: combo of apoenzyme & cofactors
ENZYME REGULATION

Enzyme level: controls enzyme activity Competitive inhibitiors: block active sites; Noncompetitive inhibitors: attach to allosteric site elsewhere on enzyme altering active site so its no longer functional - Allosteric inhibition: alters shape of active site so substrate cant bind - Allosteric activation: activator molecules bind altering shape of active site, which activates an otherwise inactive enzyme Feedback inhibition (negative feedback): final product of a series of reactions inhibits enzyme in earlier part of the pathway; slows/stops anabolic pathways when product is in abundance Genetic level: controls transcription of mRNA needed for an enzymes synthesis Transcription control: when transcription occurs and how much RNA is created Induction/Repression of enzyme synthesis by regulatory proteins that can bind to DNA and either block or enhance the function of RNA polymerase (enzyme required for transcription)
METABOLIC REGULATION

If 2 energy sources are available, cells catabolize the more energy efficient one first Cells often synthesize enzymes needed to catabolize a substrate only when substrate is available Cells synthesize metabolites they need; cease synthesis is metabolite is available
METABOLISM

Sum of all chemical reactions occurring within a cell Catabolic pathways: break down molecules & releases energy (which is used to transfer energy to ATP); hydrolysis Anabolic pathways: synthesizes molecules & uses energy (ATP); dehydration synthesis

Phosphorylation: addition of phosphate group to molecule; 3 types form ATP (from ADP) Substrate-level: transfer of phosphate to ADP from another phosphorylated organic compound Oxidative: energy from redox reactions of respiration used to attach inorganic phosphate to ADP Photophosphorylation: use of energy from light to attach inorganic phosphate to ADP 4 major strategies for producing ATP Cellular respiration (aerobic/anaerobic): metabolic process involving complete oxidation of substrate molecules and production of ATP following a series of redox reactions Fermentation Photosynthesis
AEROBIC RESPIRATION

Final electron acceptor: oxygen Glucose + ADP + O2 ATP + CO2 + H2O Product: 34-36 ATP Substrate-level & oxidative phosphorylation Glycolysis: splitting of glucose molecule Substrate: glucose, NAD+ is required coenzyme Products: 2 pyruvate, 2 ATP, 2 NADH (reduced NAD+) Acetyl CoA synthesis: 2 carbons from pyruvaic acid join coenzyme A to form acetyl CoA Substrate: pyruvate Product: acetyl CoA Krebs cycle: transfers electrons from acetyl CoA to coenzymes NAD+ and FAD Substrate: acetyl CoA Products: 4 CO2, 6 NADH, 2 FADH2 Electron transport chain: Series of redox reactions that pass electrons from one membrane-bound carrier to another, and then to a final electron acceptor; energy from these electrons is used to pump protons across the membrane Substrate: NADH, FADH2, ADP, O2 Products: NAD+, FAD, ATP, H20
ANAEROBIC RESPIRATION

Uses an electron transport system & chemiosmosis (ions flow down electrochemical gradient across a membrane through ATP synthases to synthesize ATP) -2 Final electron acceptor: inorganic, oxygen-containing ion: NO2 , NO3 , SO4 , HCO3 Product: less than 34 ATP per glucose; less efficient than aerobic respiration Substrate-level & oxidative phosphorylation Methanogens: obligate anaerobe that produces methane gas; methanococcus, methanosarcina; reduce CO 2 (HCO3) to CH4
FERMENTATION

Partial oxidation of sugar to release energy using an organic molecule from within the cell as the final electron acceptor Final electron acceptor: organic molecule Oxidizes NADH to NAD+ so that glycolysis can continue Only 2 ATP made per glucose End products: alcohols, gases, organic acids Substrate-level phosphorylation Common types Acid fermentation: conversion of pyruvate to lactate Alcohol fermentation: conversion of pyruvate to acetaldehyde (CO 2 produced) to ethanol Other types Homolactic fermentation: lactic acid only; lactobacillus, lactococcus Heterolactic fermentation: lactic acid, ethanol, CO2; leuconostoc Mixed acid fermentation: lactic acid, succinic acid, acetic acid, ethanol & gas; e. coli Propionic acid fermentation: propionic acid, acetic acid, carbon dioxide; propionibacterium Butanol acetone fermentation: butyric acid, acetic acid, butanol, acetone, ethanol, carbon dioxide & hydrogen gas; clostridium
PHOTOSYNTHESIS

Uses light energy to produce ATP & reduced NADPH Some organisms use this energy to fix inorganic carbon General pathway + H2A + ADP + NADP A2 + ATP + NADPH

Oxygenic photosynthesis: uses water as electron donor, produces oxygen + H2O + ADP + NADP S2 + ATP + NADPH Plants, algae, cyanobacteria Anoxygenic photosynthesis: uses hydrogen sulfide as electron donor, produces sulfide + H2S + ADP + NADP S2 + ATP + NADPH Green sulfur bacteria