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2.0 Generation of ATP

by: Gail Chernomorets

2.0 Generation of ATP BIOL 251

Gail Chernomorets

GPA 3.2

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Notes from 09/29 Week 5 Biol 251
Medical Microbiology
Kurt Regner
Class Notes
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This 10 page Class Notes was uploaded by Gail Chernomorets on Saturday October 1, 2016. The Class Notes belongs to BIOL 251 at University of Nevada - Las Vegas taught by Kurt Regner in Fall 2016. Since its upload, it has received 30 views.


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Date Created: 10/01/16
Notes from 09/29 2.0 Generation of ATP: Glycolysis & Fermentation Overview of Metabolic Pathways  Metabolism is all the chemical reactions within an organism  Catabolic pathways break down complex molecules and release energy  Anabolic pathways build complex molecules and consume energy  Pathways are coupled Catabolism  Energy from catabolism (exergonic energy-releasing processes)  Reaction releases energy trapped and stores ATP  Break down  Potential energy less than 0  Exergonic reaction - energy decreases from reactants to products Anabolism  Energy for cellular work (endergonic energy-consuming processes)  Reaction requires energy  Making  Need source of energy  Potential energy more than 0  Endergonic reaction - energy increases from reactants to products  make proteins, carbs, and lipids  homeostasis applies here * Both should be the same size Free Energy  The portion of a system’s energy that is able to do work when temperature is uniform throughout the system - energy from ATP  ex. active transport  Gibbs Free Energy - ΔG = change in free energy - Energy allows you to do work  Not usable energy – heat  What can cells do with free energy? - mechanical - transport - anabolism  ΔG < 0 Free energy is released (negative) - exergonic - released/liberated  ΔG > 0 Free energy is consumed (positive) - endergonic - consumed/utilized  ΔG = 0 You’re dead  ΔG = G final state starting state Why is this important?  Understand biogeochemical/nutrient cycles - recycled by organisms using as source of energy - carbon and nitrogen cycle  Investigate evolutionary relationships among the three domains - have own pathways of acquiring material  Identify potential targets for antibiotics or antimicrobials  Industrial production of amino acids, polysaccharides, organic acids  Fermentation of beverages and foods  Use prokaryotic models to study eukaryotes  Identify unknown  Ex. Zantham gum – polysaccharide (seen in salad dressing) Transfer of Free Energy - to ATP and reduced coenzymes 4 steps  glycolysis  pyruvate oxidation  Krebs/citric acid cycle  Electron transport chain  Polysaccharides are hydrolyzed to monosaccharides - organisms eat  Lipids are hydrolyzed to their individual components - into fatty acids  Proteins are hydrolyzed to amino acids - fed into the system  Fuel chemical bonds  electrons  energy  make ATP Most metabolic pathways are similar  Extremely similar in all 3 domains of life  Energy is released in incremental steps  Release more energy with many steps; one step has more heat  If all the energy was released in a single step, the cell would explode - kill cell Glycolysis  Dissolution of sugar  Occurs in the cytoplasm of almost every type of cell in every organism - most bacteria do this or something similar  Variations exists  Aerobes & anaerobes - evolved early - thought to be first metabolic pathways because doesn’t require oxygen  Hypothesized to evolved early  Nearly universal  Independent of oxygen  Glucste and other monosaccharide - 1 part requires some energy - 2ndpart leaves 2 ATP  Glucose (6C)  2 pyruvate (3C each)  10 steps **Do not need to memorize  Requires NAD+  1 half go higher; require energy - 2 ATP required - start very high  2ndhalf go lower; release energy - reduced (receive electrons) - H+ added - 2 NAD+  2 NADH + H+ - final state: very low ** Glycolysis releases energy; exergonic *Results of Glycolysis For each glucose:  2 pyruvate  2 NADH + 2H+  2 ATP produced.  Glucose is oxidized to pyruvate or a pyruvate derivative  Part I – Energy Investment - Requires 2 ATPs  Part II – Pay of - Free energy stored as 4 ATPs  Free energy of glucose is converted to - 2 ATP - 2 NADH - some heat is lost (not a big deal for bacteria) Process ATP is produced by substrate-level phosphorylation  Enzymes transfer PO gr4ups from organic molecules (Substrates) to ADP - to make ATP  A small quantity of ATP is formed by substrate-level phosphorylation  In contrast, a large quantity of ATP is produced by oxidative phosphorylation in the mitochondrion *Embden-Meyerhof-Parnas Pathway (EMP pathway)  Most common type of glycolysis  Present in all 3 domains of life - Eukaryotes - Anaerobic bacteria - Facultative anaerobes - Some Archaea, but there are usually differences in 1 or 2 enzymes  Outcome is the same  Produces: - 2 ATP - 2 NADH *Entner-Doudorof Pathway (ED pathway)  Slightly different version of glycolysis  Occurs only in some prokaryotes – bacteria and archaea  Two unique enzymatic steps  Produces: - 1 ATP - 1 NADH - 1 NADHP (phosphate group) For every C H6O12s6gar)  Occurs in several aerobic Gram (-), only a few Gram (+)  Most of the organisms that use this pathway are aerobes  Some Archaea perform ED, but others have variations is a few steps - Known as ED-like  Glucose  pyruvate make a little ATP Glycolysis continued  Many bacteria and some fungi can survive on glycolysis and humans have exploited this trait - on their own  Glycolysis nets a small amount of ATP - NAD+ is reduced to NADH - Have to regenerate  NADH transfers an e- to pyruvate or a pyruvate derivative to regenerate NAD+ - referred to as fermentation  If the terminal e- is absent, then pyruvate or a pyruvate derivative is reduced by NADH  Reduced form of pyruvate is a fermentation waste product  Glycolysis requires NAD+ - Death occurs if NAD+ is not regenerated  Transfer e- to pyruvate and reduced to ethyl alcohol or other fermentation product  Fermentation allows aerobic organisms to survive in the absence of oxygen - done in absence  Oxygen was absent in the atmosphere of early Earth Fermentation  Occurs in the cytosol (cytoplasm)  Advantage: regenerate NAD+  NADH is oxidized to NAD+ by transferring e-s to pyruvate or several pyruvate derivatives - depends on the organism  The reduced pyruvate product is excreted as a waste product  ATP generated by substrate-level phosphorylation - little bit of ATP  The E.T.C is not involved - fermentation is NOT anaerobic respiration - FALSE IDEA Alcohol Fermentation  Ex. Saccharomyces yeast  Pyruvate (3C) converted to acetaldehyde (2C) by removal of CO 2  Acetaldehyde is reduced (by NADH) to ethyl alcohol  Regenerates NAD+ Lactic Acid Fermentation  Ex. kim chi  Pyruvate is reduced (by NADH) directly to lactic acid (lactate) Pyruvic Acid   Lactate dehydrogenase Pyruvate decarboxylase NADH  NAD+ (release carbon)   Lactic Acid Acetaldehyde  Alcohol dehydrogenase  Ethanol (ethyl alcohol)  waste product Most Common Fermentations Bacteria and fungi are manipulated to produce ethanol, lactic acid, propionic acid, and organic acids From pyruvate:  Lactic acid - Lactobacillus  Ethanol CO 2 - Zymomonas - Yeasts  Propionic acid - Propionibacterium - Ex. stinky cheese  Mixed acid fermentation wastes - Escherichia - Salmonella - Shigella  Glucose or other sugars - glycolysis * All fermentation e- transferred to Goal is to regenerate NAD+ ** Will be on exam Organisms Fermentation Fermentation products Propionibacterium CO 2ropionic acid Swiss cheese Aspergillus, Lactic acid Cheddar cheese, yogurt, Lactobacillus, soy sauce Streptococcus Saccharomyces CO 2thanol Wine, beer Clostridium Acetone, isopropanol Nail polish remover, rubbing alcohol Escherichia, Acetic acid Vinegar Acetobacter Fermentation History  Humans started fermenting milk (yogurt), grain (beer), vegetables (to preserve), fruits, and baking bread - ~10,000 years ago  Archaeological excavations have uncovered wine jar storage facilities - ~7,000 years old  Ancient Egyptians had a beer tax - 5,500 years ago  Process remained a mystery until Pasteur determined that Saccharomyces was responsible for alcohol production  Latin, fervere “to boil”  Bacchus - Roman god of grape harvest, wine making, wine and intoxication - Good harvest led to big party orgy  Romans observed that mixtures of crushed grapes kept in large vessels produced bubbles, as though they were boiling  Early beer and wine makes depended on luck  If the mixture was not incubated long enough, the product contained no alcohol  Incubated too long, the mixture was unpalatable Antoni van Leeuwenhoek  Animacules  Involved in fermentation  Observed yeast in wort - sugar from grain - steeped grain juice used for beer and whiskey - near boiling water is used to get sugar out of the grain  Yeast were called ferment  Definition in 1755 - ferment put into drink to make it work; and into bread to lighten and swell it  Common belief was that yeast were not alive, but somehow involved in fermentation During the 18 thand 19 thcentury  Chemists attempted to determine how CH CH OH (3thy2 alcohol) was produced during fermentation - was thought to be strictly chemical Antoine Lavoisier  1789  Conservation of Mass  Balancing equations - mass is neither created nor destroyed in chemical reactions  The mass of any one element at the beginning of a reaction will equal the mass of that element at the end of the reaction  Amount of sugar and alcohol balanced  Determined that sugars were composed of C, H, and O - 1:2:1 ratio  Investigated the mechanism by which sugarcane juice was transformed into CH 3H O2 (ethyl alcohol) and CO duri2g fermentation  He estimated the proportions of C H O 6su12r6 and H O at the 2 beginning of the fermentation and compared them with the CH CH OH 3 2 and CO p2oportions obtained at the end - 2/3 of the C 6 O12e6e used to form CH CH OH (3thyl2alcohol) - The other 1/3 formed CO bubb2es (carbon dioxide)  Predicted that if it was possible to combine CH CH O3 (et2yl alcohol) and CO (2arbon dioxide) in the right proportions, the resulting product would be C H6O 12u6ar)  Could never explain why the ferment (yeast paste) was necessary for CH 3H O2 (ethyl alcohol) and CO produ2tion  Assumptions - yeast were not alive - fermentation was strictly a chemical process Yeast  Determined to be absolutely necessary for fermentation (1815)  Several biologists agreed that yeasts were living organisms (1835) - Better microscopes - Reproduce by budding  How they grow  Pinch off - Grow rapidly in wort  Lots of sugar  2 grape juice and one with yeast  controlled experiment Fermentation was accepted as a partially biological process rather than strictly chemical Louis Pasteur  1876  Conducted controlled experiments that demonstrated CH CH OH (ethy3 2 alcohol) was produced by yeast  Alcohol production was linked with an increase in the yeast population  **Identified that lack of O w2s important for fermentation  Respiration or without air (at the time is what it meant)  Determined that spoiled wort was caused by lactic acid fermentation by bacteria - when it did not work was a new fermentation  Two types of fermentation: alcoholic & lactic acid Eduard Buchner  1896  Wanted to understand how yeast makes alcohol  Demonstrated that the contents of lysed yeast cells could also produce CH 3H O2  He concluded that yeast contain as enzyme, zymase - response for fermentation  Zymurgy - Greek - “Workings of fermentation”  In 1929, ATP was discovered and found to be important for fermentation  In the 1940s zymase was found to be a multiple step process that we call glycolysis  Zymase  glycolysis - leads to Lactic Acid Fermentation 2 types Lactic Acid Fermentation  So NAD+ is made  Gram (+) bacteria that produce lactic acid under anaerobic conditions  Commercially valuable and used to produce kim chi, sauerkraut, yogurt, sour beer 1. Homolactic Fermentation  EMP (version of glycolysis)  1 C 6 12is6converted to 2 lactic acid - lactate dehydrogenase  Genera that perform - Lactobacillus - Lactococcus - Enterococcus - Streptococcus - Pediococcus  e- taken from NAD to NAD+ 2. Heterolactic Fermentation  1 C 6 12yi6lds 1 lactic acid, 1 CH CH 3H a2d 1 CO 2  * Utilize the Pentose Phosphate pathway and not glycolysis - functional equivalent The Pentose Phosphate pathway branches giving rise to equal amounts of ethanol and lactic acid Genera that perform - Leuconostoc - Oenococcus - Weissella - Certain Lactobacilli  Produce - lactic acid - ethanol - CO 2


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