Microbiology BIOS 3120
Popular in Course
Popular in Biological Sciences
This 393 page Class Notes was uploaded by Donald Jenkins on Wednesday September 30, 2015. The Class Notes belongs to BIOS 3120 at Western Michigan University taught by Maria Scott in Fall. Since its upload, it has received 105 views. For similar materials see /class/216865/bios-3120-western-michigan-university in Biological Sciences at Western Michigan University.
Reviews for Microbiology
Report this Material
What is Karma?
Karma is the currency of StudySoup.
You can buy or earn more Karma at anytime and redeem it for class notes, study guides, flashcards, and more!
Date Created: 09/30/15
Nutrition 11 Lecture 5 1 392420 1 1 Electron donors Energy source energy is released upon oxidation of a compound The partners that participate in redox reactlons are cruc1al The amount of energy released in redox reactions depends on both the electron donor and the electron acceptor The greater the difference between the reduction potential EO39 of two components the more free energy G 039 released What is the electron donor Electron Tower Examples of reactions with H2 as e39 donor 1 H2 fumaratez gt succinate2 AGO 86 kJ 2 H2 N03 gt N02quot H20 AGO 163 kJ n 1 1 H2 202 gt H20 AGO 237 kJ Couple COZglucose O43 39 2HH2 042 2 e Gogmethanol 038 6 equot NADNADH 032 2 e Gogacetate O28 8 e 80st 028 2 e 80427H23 022 8 e Pyruvatellactate 019 2 9 34062732032 0024 2 e Fumaratesuccinate 003 2 e 2 Cytochrome boxred 0035 1 e Fe3Fe2 02 1 939 pH 7 Ubiquinoneomed 011 2 e Cytochrome camel 025 1 e Cytochrome aomed 039 1 e gt N03 NOZ 042 2 e Nogf xz N27 074 5 e FeStFe2 0761 9 pH 2 15 O2H20 082 2 e potential Is N03 a better electron acceptor than fumarate hint what do you know about their Reduction Potentials EO39 Transfer of electrons in redox reactions requ1re carr1ers primary e39 donor initial donor terminal e acceptor nal acceptor Two classes of electron carriers Freely diffusible coenzymes Prosthetic groups attached to enzymes in CM Common diffusible electron carriers Nicotinamideadenine dinucleotide NAD and NADphosphate NADP NAD and NADP transport both electrons 2e39 and protons 2H simultaneously NADNADH is normally directly involved in energygenerating or catabolic reactions Within the cell NADPNADPH is usually part of biosynthetic reactions anabolic reactions NADH NAD REDUCED FORM OXIDIZED FORM I I Nicotinamide H H N H 1C Ho P o CH O Ribose R 0 OH OH MHZ Oxidized Reduced HO3O lt l j o CH N N Ribose Adenine OH OH Electron and H T 39 I atom carrlers HOPO Phosphate added in NADP Reunion l Enzyme reacts with substrate electron donor and oxidized form of coenzyme NAD D binding siteAttlve me Enzyme NAD Substrate electron on r Remtion 2 Enzyme l reacts with substrate electron acceptor and reduced form of coenzyme NADH ve NADH Acti binding site site d Su bstrate electron acceptor NADH Substrate Ize Enzyme and produtls Figure 511 Brock 2006 Pearson Prenlke Hall ln NAD Substrate Enlyme and produds Biology of Microorganisms 11e Energy released by the cell through redox reactions and the like must be conserved Energy is conserved in the form of phosphorylated compounds What is the primary PO41ated compound used to store energy in the cell quot2 CHO Ester N I Anhydrlde Anhydride bonds bond NI gt HltIZOH bond l N OHCH N l n 39 39 0 1 1 CH2 Cc 0 0 o o CH Hclou Ir H3C 3 o o o CHzo 0w 0 0 o Acelyl phosphate 0 OH OH OP 0 ll Glucose 6phosphule Phosphoenolpyruvnie Adenosine lriphosphuie ATP Thioester Compound 6039 kJmol bond Aa gt 30k 0 Phosphoenolpyruvate H ll H II Hg CITS CszN C CH2zN C 39SH23OTR 13Bisphosphoglycerate n Acetyl BMercapto Pantothenic acid ethylamine ATP ADP AtetyICoA Acetyl oA A6 39 lt 30k AMP Acetyl phosphate Glucose 6phosphate Figure 512 Brock Biology of Microorganisms 112 9 2006 Pearson Prentice Hall Inc Energy Storage normally present at low levels 2 mil lirnolar Two mechanisms of energy conservation in chemoorganotrophs are fermentation and respiration example E coli In both methods synthesis of ATP occurs by the energy released in oxidationreduction reactions HOW Does Fermentation and Respiration Generate Energy for Bacterial cells Fermentation ATP is synthesized during steps in the breakdown of an organic molecule CP ADP DATP a Substratelevel phosphorylation ATP is produced during respiration at the expense of the proton motlve force x Energized X x membrane x X X ADP Pi JATP Less energized x membrane ATP synthesis occurs in phototropic organisms similar to oxidative phosphorylation But light is used to generate PMF oxidation during fermentation is 111016011131quot oxygen 0r coupled to reduction anOth e l quot GXt mal of a compound electron acceptor is generated from the used initial substrate No external electron acceptor is involved Comparison of Fermentation and Respiration Fermentation 1 ATP produced from products made during fermentation process 2 ATP Respiration 1 ATP produced at expense of proton motive force 3 ATP Common biochemical pathway for fermentation of glucose Glycolysis EmbdenMeyerhof glucose to Glyceraldehyde3 phosphate No energy output ATP used No oxidationreduction rxns ATP ADP ATP ADP J j Hexokinase Phosphofructokinase A 4 plt gt 1 cm P 4 P 71139quot 511 Gflyceraldehyde 3 Phosphate iii 39j 1 3 Making ATP GiyceraldehydeS P makin ruvate dehydrogenase gpy 2 Pi Electronsgt 2NAD P 2 NABisgeduoedtoNADH 2 ADP Phosphoglycerokinase g 2ATP N0 net gain in ATP synthesis between stage I and II but at end of Stage II 2 moles ATP produced Conversion of Phosphoenolpyruvate J Pyruvate nets 2 molecules ATP Stage III Fermentation products t 2 Phosphoenolpyruvate39 P A 2 ADP Pyruvate kinase g 2 AT P Stage III eduction 2 P ruvate39 Makmg i NADH y PyruvateFormate lyase fermentation A Lactate P t ruva e 39 products NAD dehydrogenase dgcarboxylase Acetate formate Formate Lactatequot Acetaldehyde C02 lhydrogenlyase Alcohol NADH H2 002 dehydrogenase NAD r Ethanol Reduction of pyruvate oxidizes NADH to NAD allows glycolysis circuit to continue Stage III Pyruvate is the pivotal molecule from which come all fermentation products Ethanol Lactate CO2 Reduction of pyruvate Via fermentation Yeast ethanol and CO2 Lactic bacterial lactate reactants and products must balance Glucose C6H1206 2 ethanol CZHSOH 2 lactate C3HIOO3 2H Fermentation products of Saccharomyes cereV1s1ae Yeast chemoorganotrophs anoxic conditions yields alcohol and CO2 as byproducts few cells Essentials of Aerobic Respiration Transformation of organic carbon compounds to CO2 Transfer of electrons from organlc molecules to terminal electron acceptor ATP synthesis depends PMF Electron Transporters Electron transport systems membrane assoc1ated electron carrlers NADH dehydrogenase Ribo avin avoproteins FADFMN Ironsulfur proteins Cytochromes Lipidsoluble quinones NADH dehydrogenase proteins bound to the inside of CM tactive site binds NADH accepts both electrons and protons when NADH is converted to NAD passes the electrons and protons on to avoproteins Flavoproteins derivative of ribo avin accepts 2e39 and 2H only donates electrons Isoalloxazine ring I Holl O Cl l Cl Cl H2 OH H OH OH OH I Oxidized Ribitol protein bound prosthetic group 72H292H Flavin niononucleotide Iron sulfur proteins electron carriers RCysteine S Cysteine R e 39 x Fe gtFe V V V P RCysteine S Cysteine F R G t 39n S CysteineR lgt 39FCysteineR S r Cysfei nei b CHC CH2nH Cytochrome carries electrons only Pyrmle H Protein 6 CysteineS SQySteine39 Aminoacid Aminoacid Porphyrin A atetrapyrrole C20H14N4 I Cytochrome b C Substrates NADINADH E C d 4 O D C 2 q 0 5 39U G I Electron Transport Chain Generation of Proton Motive Force k x r 1 JudquotFquot 1 3H o2 xi 7 CYTOPLA H quot 7 39 52 f ENVIRONMENT r r r f Energized membrane PMF Drives ATP Synthesis ATP synthase enzyme complex 1 multisubunit headpiece F 1 2 proton conducting chal ll l61 F0 Catalyzes a reversible reaction between ATP and ADP Pi Proton Motive Force Protons cannot diffuse across the membrane crosses Via the ATPase proton channel FlF0 Smallest Biological Motor Yet ADPPi ENVIRONMENT ATPaseCatalyzed ATP Synthesis Is Oxidative Phosphorylation The ATPase motor can reverse l Hydrolysis of ATP provides torque for y subunit to rotate in the opposite direction 2 Allows protons to be pumped from inside to outside the cell creating PMF Carbon ow during respiration gt TCA Cycle o Citric acid cycle 0 Krebs cycle 0 Production of reduced coenzymes C02 H20 and ATP Carbon Flow in Citric Acid Cycle 0 Early steps same as glycolysis 0 Product pyruvate completely oxidized to C02 0 Production of AcetylCoA is pivotal NAD CoA Pyruvate three carbons NADH 002 Key A t I c A C2 ce yA 0 C4 CoA C5 Ce OxalacetateQ Citrate3 NADH 3 NADD Aconltate Malatez lsocitrates39 NADP Fumaratez FADH j C02 FAD SUCCinatGZ a Ketoglutarat z39 39 39 NADPH Succinyl Co GOA NAD 00A 7 GDP Pi 002 NADH a GTP The Citric Acid Cycle Is Key For Biosynthesis and Is Not Just For Energy Generation 439 39I 39 Ar f u u i n39 If uquotquotquot quot I lyingquot in 5 I 1v I39 I n T mwmane w m e all y 39 Eur Erica l39n ii If Jl39qt 1 E q q E dff i 1 39e ju 39 1 4 n ATPase P recharges membrane potential E Ergl membrare W I quotq n 522 a 1 L 13911 Brida me pimpinnwlatim Catabolic Alternatives to Fermentation and Respiration Anaerobic respiration Chemolithotrophy Phototrophy Anaerobic Respiration Electron acceptors other than oxygen nitrate NO339 ferric iron e3 sulfate 042quot carbonate CO3 2i39 other organic compounds Chemolithotrophy Use of inorganic electron donors hydrogen sul de H28 hydrogen gas H2 ferrous iron F6 2 ammonia NH3 User C02 as a carbon Phototrophy Two types Photoautotrophs energy from ATP used to synthesize carbon source from CO2 Photoheterotrophs organic materials is carbon source With light as energy source Many strategies exist for bacterial respiration but all roads lead to generation of PROTON MOTIVE FORCE Microbial Nutrition and Metabolism Lecture 4 Magnification Light path 100 x 400 x 1000x Visualized image Eyepiece 10x ocular lens J V Intermediate Image inverted from that of the specimen 11 6 4036quot QObjective lens x 0 w 39 specimen None Condenser lens Field diaphragm Light source in a b cauyngw a 2002 Pearson Ecucaimn inc puniishiug as Pearson Eewavm nmmgs Compound Light Microscope 1 Compound more than one lens 2 Parfocal remains in focus when you ob eetIves Microscope magni cation Ocular lens 1 OX Obj cctivc lens 10X 40X 100x Resolution Capacity of an optical system to distinguish two adjacent points as separate Resolution 1 Light must pass inbetween two objects in order to resolve them 2 The shorter the wavelength the better the resolution Resolving power depends on l wavelength of light used 2 innate property of the objective lens to gather light apertures Resolving Power 05 L light wavelength numerical aperture Refraction 1 Bending of light as it passes through object lens 2 Glass and oil have the same refractive index Goals for this portion of BIOS 312 l The chemical building blocks of cells HOW cells acquire energy Microbial nutrition Chemicals NUTRIENTS needed by cells for the construction of various cellular components and to sustain life Macronutrients needed for cell growth 1 Macronutrient large amounts a Carbon and Nitrogen 2 Maeronutrient smaller amounts be Phosphorus phOSphate sulfur Na potassiqu Nutrients needed for Growth cont d 1 Trace elements small amounts are needed metals role in function of enzymes e g Iron extremely important trace metal required for cytochromes Major components prokaryotic cells Percent of total Element weight 50 20 Minor components of prokaryotic cells Percent of Elements Dry weight S Na K 100 each Ca Mg Cl 050 each Fe 025 each Nutrients required for cell growth 1 Growth factors organic compounds such as Vitarnins cg Folic acid B12 B1 Culture Media Synthetic Media Complex Media Chemically Chemically de ned unde ned distilled H20 yeast extract pie ptonea PhOxSpih thg 7 carben and energy sourGe 39 sulfate calcium and Key metabolic terms Metabolism Anabolism Catabolism Microbial nutrition NUTRIENTS needed by cells for the construction of various cellular components How do cells gain energy Chemical compounds source of cellular energy OR Energy is Stored in the Cell As Adenosine triphosphate ATP Anabolism Biochemical process involved in synthesis of cell components from simpler molecules REQUIRES ENERGY INPUT Catabolism Biochemical process involved in breakdown of organic or inorganic compounds ENERGY PRODUCTION METABOLISM All biochemical reactions in a cell both anabolic and catabolic Cells are open systems Simultaneous biochemical reactions occur in the cell growth movement reproduction t0 The Timing and Location of Cellular Reactions are Precisely Regulated by Enzymes Enzymes are catalysts that are strictly regulated highly speci c for their target nct up in the reaction Enzymes Enzymes effect the speed at Which a reaction takes place It does not effect the equilibrium for Attributes of Enzymes 1Globular proteins 2 Catalyze between 110000 molecules per second 3 Hith speci c for their Activation energy energy input needed for molecules to react With each other Attributes of Enzymes 1 Raise the rate of chemical reactions from 108 to 1020 faster than they occur Without the enzyme rVvlv v v w y v v v v pmgress of reaction g Ace mmm mail Em ags The EnzymeSubstrate complex Enzymes interact With one substrate or speci c class of substrates The active site of the enzyme subatrates FIde um enzyme enzyme substrate enzyme complex Free Energy of Formation Gof The energy that is produced or needed to make a certain product from its constituent components Th6 fre i i rgy of elemental Free Energy Go Formation of compounds can be exergonio the G0 is negative and energy is released Free Energy Go If formation of a compound is endergonic the G0f is positive and energy is needed for the reaction to occur Free energy of formation of some 00111111011 compounds Water H20 Carbon dioxide C02 Hydrogen gas H2 Nitrous oxide N20 Glucose C6H12O6 G0f 2372 kjmol 3 944 quot O n 1042 quot 9173 quot Temperature Pressure Concentration of reactantsproducts p H Ofthe S39Olution Measured at 250C 1 atm pressure pH 70 1 M concentration OxidationReduction reactions Energy released from chemical reactions in living organisms involves oxidationreduction reactions quot Removal of an electron or electrons from a substance Reduction Addition of an electron to a substance REDOX reactions involve transfer of both electrons and whole hydrogen atoms Redox reactions involve electron donation by an ELECTRON DONOR and acceptance by an Example of Redox Reaction H2igt2 e 2 4 Electrondonating half reaction 02 2 9 02 Electronaccepting half reaction 2 H 0239 H20 Formation of water Electron Electron donor H2 02 i HZO acceptor Net reaction Rules of Redox Reactions The substance oxidized is the electron donor ie Hydrogen gas The Substance reduced is the electron Reduction Potential EO39 Measure of the tendency of a molecule to be reduced at the top of the tower electron donor be oxidized at the bottom of the tower electron acceptor reduction Electron Tower potential Examples of reactions Couple En V a w s 050 mm H2 as e danor Gogglucose 043 24 equot 2HH2 042 2 e 040 COQmethanol 038 6 e 0 30 NADNADH 4332 2 e 1 H2 fumaratezquot 1 succinatez39 COZacetate 028 8 e V a 020 AGO 86 M SOHQS 023 2 3 80427st 022 8 e Pyruvatelactate 0192 90 a 00 84052782032 0024 2 e a 4M0 39139 Fumaratesuccinate 003 2 e 2 H2 No 5 N0 H20 2 Cytochrome awed 00351 9 AG a 163 k Fe3Fe2 02 1 e pH 7 Ubiquinonewred 011 2 e Cytochrome COXred 025 1 e Cytochrome amred 039 1 6 gt NOS fNOQ 042 2 e 39 No3 12 N2 074 5 e 1 H2 2 02 r H20 FeSFe 07B1 9 pH 2 A6 i237 M 12 02H20 082 2 e5 WHAT IS CHANGE IN FREE ENERGY AG0 The energy yielded or required for the formation of a speci c molecule from its constituent elements A D AGO39 Relationship between Redox rxns And AGO free energy and AEO reduction potential 1 Transfer of electrons from a donor to an acceptor results in a change in the reduction potential Volts Relationship between AGO 39 and AE0 The greater the difference in the reduction potentials between two redOX partners the more energy released high energy phosphate bonds are the energy source of the cell CH23 COO 23 High energy Low energy N HCOH LOW energy High energy 0 P O anhydride bonds ester bond l ester bond anhydride n N OH 0 939 1quot 0 CR O Phosphoenolpyruvate O IIT39v 0 0 O CH H30H l Hac C ONE 0 o o o CHz O O O o Acetyl phosphate OH OH Adenosine triphosphate ATP Glucose 6phosphate Compound 60 kJmol High energy Phosphoenolpyruvate 516 13 Bisphosphoglycerate 520 Acetyl phosphate 448 318 M t ADP 318 erca o 39 39 Acetyl Ethwami e Pantothenlc acrd Low energy AMP 142 AcetVlCOA Glucose 6phosphate 138 ATP AdenosinePPP P phosphate group phosphoanhydride bond Breaking of PP bond results in release ofenergy er kealm ol of Summary of ATP Hydrolysis Phusphuanhyd dn bonus III x F39Im mnhmmm mud Mgr v Hamlinn I Hvdmwum x I 7 MN 3 Math Di 0 J Q x V H I y H a 13 p g p u p n EH 0 maulum 2 ATP nunu m 39 39 MG 2 13 hcan39mmj D 0 D F hnuphn l granp DH dElI39IGBI B Kquot r v quot r a Fl MP J J y 39 w Earn1 3 139 rr39rn39rrs Funurn w rum111 1n Irvlr39r39z39v 7 rmnm Adenosinetriphosphate high energy bonds Free energy released post hydrolysis of PP is transferred to other molecules Adenosinemonophosphate AMP Bond The phosphoester bond of AMP is a lower energy bond Coenzyme A another highenergy compound AcetylCoenzyme A acetylCoA with a sulfoanhydride bond 39p1 oduots derived from hydrolysis drives of ATP gt Concentration work or ElhanolCO V a F Elaclricalwolk Mechanicalwnrk Bioluminescenl work a Anaerobic conditions r Synthetic work gt Concentration work 1 Eledrlcal walk Bialumlnnwam work Heat up Aerobic conditions mm a mm vs rmquot Emmmm m pmmnnas 5mmquot oummmns Welcome to BIOS 312 Microbiology Instructor Dr Maria E Scott TA s Pietra Kohlcr Microbes Consist of OBacteriaArchae2Hlt OAlgae OFungi OProtoza BACTERIA OSinglecelled organisms OVisible under the light microscope 9N0 nucleus i1 1gular bacterium m alffig ffiMUN ii ilii 1 xV Bacterial Colonies on Agar Plate Archaea QSame size and shape as bacteria OOften isolated from extreme environments OTheught to be eldest life forms ALGAE OMicroscopic and macroscopic OContain nucleus OPhotosynthcsis OSinglc and multicc llular FUNGI OMicrcsccpic and macroscopic ONuclcus OSinglc and multiccllular O Sinular Yeasts BUD SEARS Mold on Apple PROTOZOA OSinglecelled ONucleus OSingular protozoan VIRUSES OAcellular entities ONeed living cells to replicate OVisible under electron microscope OSingular History of Microbiology OVan Leeuwenhoek OPasteur Antonie van Leeuwenhoek van Leeuwenhoek 1632 1723 ODutch amateur lens grinder OFirst to observe microorganisms OAnalyzed rain water scraped microorganisms from his teeth OCalled them animalcules OReportjexd to the Royal Society of UH HAW i E I A4 I IEIE Iil lii V Louis Pasteur 18221895 Louis Pasteur Olnvented process of pasteurization ORefuted spontaneous generation OSWanneeked asks oVaeeinated life stock and people Air frce DU pen and g 74 44 Nunaterile liquid Neck 43f flask Liquid steri ized named int flask drawn nut in by heating ame DUEt Erld mi r rg ni mg nned in band quit angled i wiy many years Hash tipped ee Miereer enieme miereergeniemIeden new in iquid duet eeeteete sterile liquid 444 4 I 1 4II 4 T4 w 4 44 4 4 4 44 4 444 444 44 444 J L44 44444 4 4 w 4IE 4 m 4 4 4 4 44 I 44 U 44 4 4 4 4 4 44 4 44 4 I 4 444 H x l 4 I 4 w 4 4 I LN 4 w l 4 414 4 4 I4 44 y E 2 1 I 4 4 4 4r in I 4 44 4 E 44 444 4 4 44 4 4 4 t h M 4 4 4 l4 4 4 4 I 4 H Eu 1 E a n EH H u H 4 m E 44 n4 WL m4 4 4 HE E E jaw Robert Koch 18431910 Robert Koch OWas the rst to use agar plates to isolate pure bacterial colonies ODeveloped Germ Theory of Disease I l u n u n n in IM uh wk 139 uh wk I v I I 3 39a39 l i uquot 39I39 7quot r 3 I I p I h I 5 r4 u c 7 39139 I v I c n I q kv I Fv39l 5 I I I U a He reniems preeth e J v 7 Streee eger plte 7 11 Li 7 m r J H 39 i I H I 1 u I quot r H I 1 J w E 7 n 1 1 r z e 4 x 1 r A J r fl Ill 1 4 I n 39w 1 l I 7 7i quotJ E gt U r H j Ii 739 l j 39 w r H 1 J L L1H m L H b 1 UH H t J 1 MN m quot U m r U 3 A r n v fre m w e 39Tl Ier I l I J 391 LI U U I m 1 pi AHA 1quot 1 JUN a w 7 quotIN I He d A FF n 1 H quotId 14 11quot uj l J I 3994 L 4 quotIX 39 139 i m H IEEE we UT 7 V Pm 39 m 1 7 HJ39JJT H e 139 39 e 7 x gt 1 I Ml I V V m 7 gt gt quot quot 39 39L In u 1 r w 1quot xi 39 739 l l 1 x quot w i e 1 39 Y J r U I g r 39 I39 I eeulete healthy enimel with eele ef eueeeeted pethegen Iv rv q n o n my rug a II u a It p 0 V1 z on u cl I I rill quotnL shmlllt39lll 394hl D I I m I ivvn i I n I t VI Hmwa hm m fisauee gamma and sarve y mi r py Lart w gulture FLIFE Eul39tura must In Earn as fare rganism Koch s Postulates O The same pathogen must be present in every case of the disease The pathogen must be isolated frm the diseased host and ltZgtThe pathogen from the pure culture must cause the disease when it its inoculated into a healthy susceptible laboratory animal ltgtThe same pathoen must be isolated Microbes in our Daily Life Out of sight out of mind However oFermented beverages Fermented food Microbial Revolution O Germ Theory of Disease OVaceinations OAntimicrobial Compounds Microbial Revolution OMolecular Biology and Genetic Engineering OBacteria important for oUnderstanding processes of replication transcription translation lt Cloningl ltgtDiscovery of Microbial Diversity First focus on E coli and yeast Then use molecular methods to assess biodiversity the real biodiversity is in the microbial world bioremedi ation Next Microbial Revolution is Up to You OFewer than 02 of microbial species have been described OMetabolic diversity OYellowstone Park of pollution What new diseases await us in the 21St Century Why Study Microorganisms OThey are our ancestors QLifesupport system of our planet OKey to life on other planets O astrobiology OSomeftiimes our enemies Lecture 3 Cell Structure Cell wall of grampositive bacteria Teichoic acid Wallassociated protein a Lipoteichoic acid 3lepticloglycan Cyloplasm39c membrane Cell wall of Gramnegative Bacteria Out a Lipop olY39 R sacchalride Outer a membrane 1 1 quot J H 7 I 7 if M rr A a LI p0 prot 5 I l E Phosphollpld Periplasms Peptldoglycan We 3 3 25 n quot39 quot q azeiitrr e39wr QQa Cytoplasmic l VI membrane 1 6w a m In Grampositive Gramnegative Peptidoglycan Peptidoglycan Membrane Membrane Peripasm Outer membrane Iipopolysaccharide and protein quot93 WWWrockefellereduvafcells Later membran Periplasm Cytoplasmic elm Nb ra me 39dgE EHEI I n 1139 u quot4 WV 1 h J39Mi quot w A If quotin I IMM L39JJ WH V 4 39 1 1 Iquot r Slip 4 Q5 n d 13 Terryr B 1 I 39 rr f IV39 I t L 39 gt7y r Mira V bquotghmm 39 39f quot 39 quot 39 LLMJ 5 Em yrigi t Erin E IPEI Pearsm E iu atil r fl Inna mblishi g 35 Pearam Eneraiamm Bm nrmngs Glycan backbone 39 C E lnterbridge L Alia LAla I y I DGluPeptldes DGIUNHZK Dl i nP DAla L ITyse 3 H xnAla DAP DaAla a Escherichia coli DGlLlINH2 gramnegative ma Staphylococcus aureus grampositive quotI5 1 Peptide bonds 39L r M Glycosidic bonds 0 Cogywigh2031 F39Earacm Educatiam 1quotle publishing Pearsor m Benjamin Cur nmings r I I I 1 Lysozyme A dig sts Wan I I I I 39I l C i39 II I 39 I I I I I I 7 I 1 I I I 39 39I39 I l39 quot 39 l I I I I II 39 I I I I rI I I quot39 i I 39 N I I I II I i I I I I I V V I g y I A v I I I I I I I I I h 5 II39 I I II 39I I l l I I r I I 39 g r r I quot I 39 g I I g l I quot g a I I quot I I s I v I I I I I I I r I I I d I b If I I 4 I I I I I I l I l I i I I I i Y 1quot J g h I w I I i 1 r I n xl d I u W I l IIDM I I I I I a s r I I I I 39 I v r I I I I a v b L I I I I I 41 I I I quot 4 quot I I I quotquot a 39 I 39 I 39 I 39 I 39 a 39 I 39 V e I 39 II I 5 39 T I 39 I I t 39 39 I I I 1 r T I I V h J 39 J l I I r a 39 l 39 I I a 39 39r 9 b I 39 In a39 II n 39 s I u L I39 quot i In I l39 a r I 39 a I I I 3 a L I I A397 I II I r 39 1 II d 2 I I I I 39 I 39 l I I a a I I I I quot I r 7 I I m I a I I I I I quotI 1 I Ix I 39 I I I I l v u I u s n r1 w I I A l l p I I I 39I 3 l y I l l 1 i I I I I 39 I I I I I I I 39 I 1 39 I i I I 1 I n 39 4 I If r n I I I quot r II A I 9 39 t 39 IrJ 39 39 rr 39 II I quot I 39 t I 39 q I l x quot I 39 I I I quot III I 39 I I I I 39 a I I I II I I I I I gtI i r I I In I 1 I I I I I I r I I I quot I I I I I I 1 e I 39 I 39 39 39 I I 39 39 I I quot I I I I 39 I I 39 I n 39 I h I I I h I i m I k l I I I h n I z h I t A I I I39 I 39 I39 I r I 39 g A i I I I I I I I a I I Iquot I II I 39 I I I I 1 39 I I quot l I I I 1 9 i x I 0 1 I I q I I II p q A l I I I w I I I I 39 I I I I I I 7 I I a I I quot a I 1 quot I I I I 39 I r I 39 r 39I I I I 39 39 1 I 39 39 r I 39 z a I 39 z c r I I I I 39 I I I I 39 1 I I I I I I 7 l I I I I a a k u I l I I 1 A I 39 39 a 39 39 I I 39 I r I I I I II I 1 39 I t I II r I quot v l II II I I I I39 I I II I I 39 II 39 39 l 39l I 39 I I 39 A I 4 z n 7 x III 1 II I I I 39 I 39 r 1 1 I 39 l I r u l I I I w I I v Iquot I I I A quot t i v I Iquot I l I I I l i I r I a r39 l I39 1 39 I I39 I I I J v39 39 I Ib r s I I I M I I I I I 39I av a J 9 V39 g 39II m I I F I 39 I 4 h I 39 I I I I I quot I I gt A I I I l 39 39 I 39 r l I 39 r I I I I A quot I In I I I I l I I i I I y u I 6 9 g I 393 f l I quot g I V quot g I quot I r F I l I 1 l I 1 I I ll I I o I 39 I I II I v II gt I 1 l I x 39 1 f I39 I I39 4 I y I 39 V39quot 39I I 39I 39 I V39 1 I I 39 39I I IIn i I qIu F 39 I 3 e 39 In 39 quot I II 39 a I a I I I l I I 39 I i g I 39 I I 39 l I I l 39 I I I I I I I F I r i i I I F l i r J 1 I l r 39 a l l V 39 I In I 939 A I I I I I I I39 quot JII III I I Iquot q I 1 quot II I I quot II II I I I I I h I 39 m 1 aIl I II I I I II l V PI 3 A a I 39 39 39 I I 39 I I I I I I l I I u I r I I I I I ISIO u I I I I 39 I I I I I I II I I I x I a I v I I M 1 II Differential Stains ODO not stain all cells the same OGrarn stain ODifferences in cell wall Most important stain for differentiation of bacteria 3 Basic Shapes of Bacteria OCcccus pl cocci spherical OBacillus pl bacilli rodshaped r Other Forms OVibrios commalike OSpirochetes contain axial lament Shapes of Bacteria Coccus 3 H h Stalk yp a Budding and appendaged bacteria Filamentous Further Characterization ODiplococci pairs OStreptococci chains gtM0n0trich0us v single polar agellum m ltgtAmphitrieh0us agella at each end of bacterium Lophotrichous tuft of several agella at one end Peritrichous agella distributed over the entire cell Flagella OFilament consists of agellin OHook OBasal body Biosynthesis of agella Filament synthesis Late hook Early hook Cay Motor I I MS ring proteins P mg L ring gr Peptidoglycan j Cytoplasmic membrane Flagella OElongate from tip OFlagellin subunits pass through hollow core of agellum and are added to the terminal end of the growing lament Flagellum anchored DireEti If I D Eli Enlargement 3 El lrEgi r l utar 39 quotquot quot E1 LPEJ xiffilfff 3 Z W74 m39 EHE a IQ g u if 55M i l l l mg r rau WERE at r r r r r r r r r r r l gt I 39 39 h r ma u i Iwly I I N I If a rigiagl L lii 1 7 FE F E plasm Syl lplasmic he a mem mna l 3 Mal rtir 1MB Dll39ijl il39 ifli pr leima matEur switch into cell wall 39Iagellum Peritrichous agella movement OMovement by rotation propeller like 0 Tethered cells rotate OCounte rclockwise bundled run CIOCkWise tumble Tumbb ageWa pushed apa CW ro tati n Bundled h Hagel a DEW rotation Flagellla bundled SEW rotamtim Chemotaxis OGradient sensing of chemoattractant or chemorepellent ORun towards attractant OChange direction away from repellant N0 Attractant Random run and tumble b Polar reversible flagella 7 7 7 it a CCW rotation CW rotation Polar unidirectional agella w Cel gt f stops CW rotation reorients CW rotation Cell Appendages Fimbriae adherence ne and shorter than agella Pili DNA transfer conjugation Comparison of mbriae and agella Comparison agellum and mbriae A flagelmmm Bjimhriaa t h0t W W 5 us alummu FIiJI a r illsruld quotHIka 4 iIIIIici P if I II r c e auv w agalns gt defenses I e n r a b M mm 6 Wm or DC kp e b W m 10D 1111 may confer prote 111111111116 Funct Capsules Glycocalyx Capsules l rigid matrix polysaccharide protein 2 antiphagocytic 3 adherence to host cells 4 protects from dying role in bie lm fermation 2 JUL 13 m 1 quotquotH1H c 39 L V I 7 9 Slimelayers Similar in function to capsules 1 less rigid more exible diffuse mat of polymer Poly Bhydroxyalkanoate Inclusion bodies Function a storage of energy b reserves of structural building blocks poly39B39hydI OXybutyrate PH PolyBhydroxybutyrate PHB rm CH 39 39 1 CH O CH CHE subunit TEM of Rhodovibrio sodomen s w lt3 9393 CH C CH Bcarbon Monomer Poly Bhydroxybutyrate F R Turner and M T Madigan Cell granules cont d Polyphosphate granules inorganic phosphate Energy source elemental sulfur accumulate inside cell Sulfur granules in T hiomargarita Magnetcscmes symmetrical Crystal particles of Magnetite Fe3O4 1 respond to geomagnetic eld Magnetotactic Bacterium 7 Whmwwmwg J gas VESiEIBS at a cyanobacterium Ferdinand Cohn Endospores OResistant OHeat ODrying Ragdiation 9 Bacillus subtilis endospore stain ABM MicmbeLibrawnrgEIJChamberlain 39439 6 pyrlg 39r M E 20339 4 51 H I y 1 v r E1 Pearsorr Education 1 l39 publishier as Peargm Benjamin C U 1 H S Pankratz T C Seaman and Philipp Gerhardt m m i r E Core wa Spore coat xosporlum b Mature endospore Kirsten Price l megetative cell V m n n I I ihDevelopin39g spore Sporulating cell 0 Endospores OTriggered by nutrient depletion 9Complex series of events occur QOver 200 genes involved O Takes approx 8 hours pores erminate rapidly Dipicolinic acid p01ymer f1 39OOC N COOquot Ca crosslinked 8 f1 f1 lt Ca39OOC N cooCaooc N COO39Ca gt b Carboxylic acid groups Microscopy Light microscope bright cld phase contrast Confocal microscopy Electron microscope Variations of Light Microscopy OBright eld ODark eld OPhasecontrast OFIuorescence Magnification Light path 100 x 400 x 1000x Visualized image Eyepiece 10x ocular lens J V Intermediate Image inverted from that of the specimen 11 6 4036quot QObjective lens x 0 w 39 specimen None Condenser lens Field diaphragm Light source in a b cauyngw a 2002 Pearson Ecucaimn inc puniishiug as Pearson Eewavm nmmgs Compound Light Microscope 1 Compound more than one lens 2 Parfocal remains in focus when you ob eetIves Microscope magni cation Ocular lens 1 OX Obj cctivc lens 10X 40X 100x Resolution Capacity of an optical system to distinguish two adjacent points as separate Resolution 1 Light must pass inbetween two objects in order to resolve them 2 The shorter the wavelength the better the resolution Resolving power depends on l wavelength of light used 2 innate property of the objective lens to gather light apertures Resolving Power 05 L light wavelength numerical aperture Refraction 1 Bending of light as it passes through object lens 2 Glass and oil have the same refractive index Basic Unit of Life The Cell Hallmarks of Cellular Life Environment gt 1 Metabohsm Uptake 0i citemicaia limit the environment and elimination if wastes intn ll IE anvimnmnnt The nail is thus an Opt2W Systnm 2 Reproduction Chemicals from the environment are turned into new cells under the direntinn if preexisting cells 3L Difmirentiatinn Formation of a new cell structure such as a spare usually as part of a cellular life cycle Hallmarks of Cellular Life l 3 4 Commun1cat10n l g 7 Calls cammmfsare 0r interact J 1 l f primarin by maans Df Chemicals l l f 1 which are released or taken up i 5 Evnlvutlnn i Cells evalve ta display new Cx biolugiaal propertias Phylogenetic l trees Show the evolutiunary Ancestral C9 relatiunships betaweer cells ALL CELLS OReproduce ORegulate uptake of nutrients and remove waste ORequire energy oElaborate sensin systems that oNucleic acids DNA RNA 6Proteins ltgtCarb0hydrates sugars QMin39e r lsh Cells OConsist of OCell membrane OCytoplasm OGenetic material oNueleoi d or Prokaryotie Cells versus Eukaryotie Cells trnkaryutic cell Cell will f Cw g lasmi mew rane energy pf d ti m FA Ri b snmea quot1thF Eil ed pmtein A EHDWE 51fnth55 nu Hamid ukaryotic cell ytoplasmic embrane l KH Golgi body protein localization E Endoplasmic reticulum studded with ribosomes protein synthesis Mitochondrion energy roduction a R Nuclear membrane Prokaryotes Eukaryotes O No nucleus O 708 Ribosorne 9N0 organelles O Motility 0 Circular chromosome 6 Nucleus O SOS Ribosorne O Organelles O Mitochondria O Chloroplasts 9 ER and Golgi 9 Motility o Macrophage destroying Bacteria WWWlifenthuedutWlslew Cell Wall Cell membrane OCell rigidity ORole in cell shape OPrevents rupture OfeeH OSemipermeable ORegulates ow of nutrients OConsist s of lipids ill 393 a Hydth T gi n Thin section of Neisseria gonorrheae Cytoplasmic membrane lam VD mm M I UT 1 mm H 4 5 31 1 x 1 x x A quot39 1 E a I A Ti 1333 H W I 4 LE I w L m n g I Irgp h b I I m g i f Tf Fl WWW1ifenthuedutWlslcw F s rr ahilitg Barrier Frau ms leakag arid fU i ti E as a gammy hr tjra spmt f r39iutriar its intr and nut f H1 cell quotii1 a I Pr igin An hnr Sim nf many mtaims inmluad ir1 transprti i ai l fg li and chamntaiis 7 f 39r w in i1l E454 Enargy EEi IE W ti r l Ema crf ganaratinn and use fquot the warm matinee fame Cell wall consists of OPeptidoglycan bacteria OChitin fungi Peptidoglycan Murein ORepeating Disaccharide subunit N acetylglucosamine N acetylmuramic acid O39Riidity amino acid cross NMeMglumsamine a Na Acetyhmulramic acid M l Di12 l l EHE UH rm 39 39 quot 39 rm WM 1 H 3 H glycosidic bond W4 I H Flinn connects the sugars D g 0 HD I bHi C S D Lysozymasensltlve EH3 CH3 imam HBCEH L NEU IM NE l CHQ GHE EH EDOH NHE DGlLJtamac acid I H0013 C CH2 CH2 EHE SH i1 Mesodiaminopimelnc g A anme acid HSG CH CDDH x Interhridge may he EEUEI39EL additiunal aa39a here D ala H P tetrapeptide D glu 1 consist of glycan Chains of alternating residues of N acetylmuramic acid M and N rzacetylglueosami eg G linkediby 1 4 g1 39coSidicbondsabetWeen theiC1 of and The DisaccharidePeptide Subunit in Peptidoglycan WELL WALL AND CAPSULE BIOSYNTHESIS 3 Lzlanine Dglulamate Dalanine CH3 Lalanine CHfiCHfioH CHZ CHZ NHZ L diaminobntyric acid CHZ CHZ COOH Lglutamic acid CH erIlziNI Z Lornithine CHZ3 CH2 NHZ Llysine coon Gigi CH NHZ Lhomoserine LLDAP and mewDA P a Diaminopimelic acid b Lysine Grampos and Gramneg bacteria different cell wall composition OGram neg OOne layer peptidoglyoan oOut er membrane in addition to inner membrane OGram pos Omany layers of peptidoglyoan OWith lipoteiohoio acids Cell wall of grampositive bacteria Teichoic acid Wallassociated protein a Lipoteichoic acid 3leptirclogilllycan Cyloplasm39c membrane Cell wall of Gramnegative Bacteria Out a Lipop olY39 R sacchalride Outer a membrane 1 1 quot J H 7 I 7 if M rr A a LI p0 prot 5 I l E Phosphollpld Periplasms Peptldoglycan We 3 3 25 n quot39 quot q azeiitrr e39wr QQa Cytoplasmic l VI membrane 1 6w a m In Grampositive Gramnegative Peptidoglycan Peptidoglycan Membrane Membrane Peripasm Outer membrane Iipopolysaccharide and protein quot93 WWWrockefellereduvafcells Differential Stains ODO not stain all cells the same OGrarn stain ODifferences in cell wall Most important stain for differentiation of bacteria Step 1 Flood the heat fixed smear with crystal violet tor 1 min All cells purple Add iodine solution for 3 min All cells remain purple Step 3 Decolorize with alcohol briefly about 20 sec Grampositive cells are purple gram negative cells are colorless Counterstain with safranin for 1 2 min Gram positive G cells are purple gramnegative G cells are pink to red 3 Basic Shapes of Bacteria OCcccus pl cocci spherical OBacillus pl bacilli rodshaped r Other Forms OVibrios commalike OSpirochetes contain axial lament Shapes of Bacteria Coccus 3 H h Stalk yp a Budding and appendaged bacteria Filamentous E Enli cell Further Characterization ODiplococci pairs OStreptococci chains gtM0n0trich0us v single polar agellum m ltgtAmphitrieh0us agella at each end of bacterium Lophotrichous tuft of several agella at one end Peritrichous agella distributed over the entire cell Flagella OFilament consists of agellin OHook OBasal body Biosynthesis of agella Filament synthesis Late hook Early hook Cay Motor I I MS ring proteins P mg L ring gr Peptidoglycan j Cytoplasmic membrane Flagella OElongate from tip OFlagellin subunits pass through hollow core of agellum and are added to the terminal end of the growing lament Flagellum anchored DireEti If I D Eli Enlargement 3 El lrEgi r l utar 39 quotquot quot E1 LPEJ xiffilfff 3 Z W74 m39 EHE a IQ g u if 55M i l l l mg r rau WERE at r r r r r r r r r r r l gt I 39 39 h r ma u i Iwly I I N I If a rigiagl L lii 1 7 FE F E plasm Syl lplasmic he a mem mna l 3 Mal rtir 1MB Dll39ijl il39 ifli pr leima matEur switch into cell wall 39Iagellum Peritrichous agella movement OMovement by rotation propeller like 0 Tethered cells rotate OCounte rclockwise bundled run CIOCkWise tumble Tumbb ageWa pushed apa CW ro tati n Bundled h Hagel a DEW rotation Flagellla bundled SEW rotamtim Chemotaxis OGradient sensing of chemoattractant or chemorepellent ORun towards attractant OChange direction away from repellant N0 Attractant Random run and tumble b Polar reversible flagella 7 7 7 it a CCW rotation CW rotation Polar unidirectional agella w Cel gt f stops CW rotation reorients CW rotation Metabolism and Microbial Growth Lecture 397 i i 1 Figure 611 Breek Bielngy ef Micmurganisms 1 He J 9 2006 Pearson Prentice Hall s Mn Assembly of FtsZ at the divisome With other proteins that participate in cytokinesis Sellrnssemblv mm at cell munquot E E E Minute 3 k Cell wall 1 7 V In I r V 39 3 a 1 u 7 V 39 V 2 quot1 I hi4 a l 1 2 I membrane 7 7 7 i 7 7 7 my h Jgga v 1 M Cytoplasmic l J N ucleoid MinE Divisome complex o39 Z g Jup r39rf Hf 339 ea gar jiIJrJafzi3r m 1513 s 39ensudr m vajurr39lmii PquotL1 IUCII JF39 rltWEr I1 blll1393 F d quot IEI f Inquot 1y FtsZ mutant showing lamentation Gabe McCool and Maura Cannon University of Massachusem Amherst FtsZ is dynamic assembly and disassembly of the cytokinetic ring allows progression through cell division cycle Peptidoglycan synthesis critical for productive cell division FtsZ ring 039 e W Wall bands I Growth zones I 4 I r I V I V 4 I HOW do you get the N acetyl lucosamineNacetylmuramic acidpentapeptide peptidoglycan 1 EH3 C3H3 I3H3 Bactoprenol undecaprenolphosphate C55 I Growing point of Peptldoglycan 0er wall Cytoplasmic membrane Out mm 6 up my mg Bactoprenol Transpeptidation rxn crosslinkage of two peptidoglycan chains l 1 Target of penicillin f gt ENERGY Mg L Ala DGlu DAP DAla DAla RELEASED Transpeptidation DAP DGIULAIa39 D Ala m M LAla DGlu DAP D Ala DAP DGlu LAIa 7 DAla K9 5 Population Growth Doubling time same as Growth rate calculate as Phases of Bacterial Growth Lag Log Stationary gelath Bacterial Growth Curve E ElmNth phases HEB Lag Exponential Statmnaw 90 a E J 1113 quotquotquotquotquotquot I quot39rargia y mg 5 30 npticat density j E 39 050 i m f 3 TD l E 4325 250 B D 3 50 e H Time Measurements of Bacterial Growth QDirect microscopic count OViability serial dilutions OTurbidity optical density spectrophotometer u a Sighr F lt 39 2 39 PI n V I 39 h k V g 4 r i i 7 W 39 z 1 P z I w 7 a l V p E j 7 IV 1 a fatal Er s E 7 uwi39r39 v kur39vrquotrt r Is wr 11 I Vr m I 5 q 1 L k n 1 x h 3E E quot1 I n nz J r I Zn iv E m i i m 1mm 1 D3 he mm uteri 1H Iiig Flame 1mlll l J samples J i e 159 1W 139955 E 1H5 FlatE iMut Eifgarnisrns ier m mummi actwr inf mmgnal gamma M Surfage colonies CFU Colony Forming Unit Sub q wca gramme Surfaga nlmmes CFU Colony Forming Unit Caveat achieving accurate cell counts from natural samples such as soil and water Phutoceli measures Pa rd r cantammg 3 E 0 cells E unscattered 7 h light 1 Umi image Unsca emd 1 Spectrophotometer 9 0 light r optical density DD Bacterial cells scatter light 3 photonHater passing through the suspension g r em 1 Klan unit DDIUEDP What is an example of a steady state culture condition Fresh medium fr m resewir 5 3quot 1 Eterile air 3 ulnar gas v rur 5 Chemostat Flawrate rugulatur Gaseuuu headspai CuturE weasel quot Culture verfnvlr Effluent cntaining miurubialcels quotaw5 l Environmental factors that in uence bacterial growth Temperature pH Water availability Cardinal temperatures OEach Microorganism OMinimum OOptimal Different classes of microbes OPsychrophiles Oopt 15 OC OMesophiles 02540 OC opt 37 OC OThermophiles 955 060 QC OiHyperrtheerph iles I i I i I i I i I i I i I i i I i I i I i I i I i I i I i I i I i I i I i I I I i I I I i I i I I I I Ii SSWiiiIISWIIIIEI39EW39l iIISHI3930395illIICIHIIHI GelllIF l illl 39l 53911I3lquot quot a I I I e I l I a I I I I a I l I I I I II I g 39I I II a E quotI a J I n i a gutu 39 m u 3939 quot 39 39 quot 39 nHUM2quotquotiniIil ll 39l I IIIIIIIIIIII III IIolquImI I alw l 39 III3939I lt IIIII II u E4quot 323quotquotquot39 1 II IIl r quotlll IIIIII i IIII I Mquot39 I I lizi J 39 Ilt39uu I I an a may i 39 2quotquot II v IllaIn l 4 m 039 u I I quot JV uquotr I I n quot I phylumIa u I 1 I I ll IIII n I III I I I IIIIIII39IIII aiIaIIIaI 39quotquot 39IIIIIIIIIIIIIIIII quIIIIIII I I I I I I II I 9 A U ll I 3 a I Q I l 3 a i H i39w w m im39afE 2 E I pEI I MWF 31 Extremophiles Psychrophiles Psychrotolerant Thermophiles Hypertherm ophiles Psychrophile Chlamydomonas algae found on snow elds optimal 150C or lower Two samples of microorganisms found in polar ice Credit Richard HooverNASA Website Psychrotolerant organisms Can grow at zero degrees Celsius but optima is between 20400C How do Psychrophiles adapt to their environment Survival strategies lipids mucus coat greater amount of alpha helices proteins less Extremophiles Thermophiles 7 W L V7 1 339 l39 7 V us iquot39 39391 g V r U 7 7 3m rn 1 1 Equot T Extremophiles Thermophiles optimum growth above 450C Hyperthermophiles growth above 0 C Chemooranotrophls an chemo thotrophsi Red rocks result of Sulfalabus Arggaea arples a 7 i1 quot r EL a I 7 l i f 7 m V IHj J 39 m 1 7 Jr 7 EH IL 4 H E73 i g 7 Advantages of Being a Thermophile T hermus aquaticus famous thermophile Molecular Modi cations Indicative of ThermophilesHyperthermophiles Proteins Presence of various solutes stabilize proteins di inositol phosphate quot x contain fatty acids in their membranes side chains composed of of the vecarbon I I Comparison of the Ester and Ether Chemical Bonds found in Lipids Ester linkage Ether linkage Isoprzgenge carbon hydrocarben Diglycerol Tetraether Biphytanyl C 40 hydrocarbon 5 i i fi f 1441H 1 1 J Membrane protein s zz39aszz m s M W W Diglycerol h c Lipid bilayer d Lipid monolaver tetra er Microbial Growth pH OAcidophiles low pH ONeutrophiles pH 7 OAlkaliphiles high pH 1 NEUTRDPHILE ACID I 39 ILE 5E HmMAE GREJWTII LIIImlunzh39lr39inH D J D FIG I D p H J TV T hiobacillus thiooxidans Obligate Acidophile OMinimal pH 05 OOptimal pH 2028 OMaximal pH 46 Alkaliphiles Habitat High pH Soils Nonmarine microbes soda lakes high carbonate soils Example Bacillus All Organisms Require Water for Life The cell is thought to be in positive water balance Water tends to diffuse into the cell toward hiher solute Metabolism and Microbial Growth 6 Fermentation occurs in the absence of usable terminal electron acceptors ATP is generated by substratelevel phosphorylation example Glycolysis Respiration Uses molecular oxygen or some other electron acceptor ATP is produced by oxidative From which method fermentation or oxidative phosphorylation of energy generation do you get more energy produced Brie y explain Our answer in of quot the of the nal Q Why does glycolysis require that pyruvate be reduced to fermentation product Explain in terms of redox balance NADVNADH balance why important Living systems need mechanisms to 1 maintain a redox balance to conserve energy required for incorporating carbon and other nutrients into the cell biomass Conservation of energy requires the participation of electron carriers eX NAD and FAD Exempli ed in the electron transport arranjernent within membrane Electron Transport Carriers Redox Enzyme Cofactors FlaVin mononucleotide FMN accepts electrons and protons oxidized passes electron transport chain Flavin adenine dinucleotide FAD accepts electrons and protons oxidized passes electron transport chain Flavoproteins are oxidoreductase enzymes enzymes that bind FAD or FMN cofactors and accept 2 e39 2H but donate only electrons to the next carrierquot in the chain Electron Transport Chain Generation of Proton Motive Force example 4 397 pef 7 52432 if EIIVIRONMENT y weakest reductant generates proton motive force IL kinetic energy to do wor Proton Motive Force Protons cannot diffuse across the membrane crosses Via the ATPase proton channel CY I OPLASM ENVIRONMENT t 6 Y r O t m 1 a m g 0 1 O 1 B t S 6 U m S n 0 m1 F F 1 subunit catalytic complex converts Pi to ATP cytoplaSm T N E M N o m V N E 12 l I How the ATPase motor works 1 Movement of protons through the FO subunit makes the C12 subunits rotate 2 Rotation of the C12 subunit creates torque that is picked up by the ye units How the ATPase motor works 3 Twisting of the ye units causes conformational changes in the 3 unit allowing binding of ADP and Pi The ye units prevent rotation of orB subunits but allows conformational change of B How the ATPase motor works As the 3 unit returns to its original state ATP is produced Approximately 3 to 4 protons are consumed for each ATP produced Generation of ATP Via ATPase OXIDATIVE PHOSPHORYLATION The ATPase motor can reverse 1 Hydrolysis of ATP provides torque for ye subunit to rotate in the opposite direction 2 Allows protons to be pumped from inside to outside the cell creating PMF Strictly fermentative organisms can not do oxidative phosphorylation don t have electron transport chain chains still have ATPase used to generate PMF Inhibitors of electron ow and PMF eX carbon monoxide CO cyanide CN bind to cytochrome Uncouplers prevent ATP synthesis without affecting electron transport lipidsoluble substances like dinitrophenol make membranes leaky destroys PMF and ability to drive ATP synthesis Carbon ow during respiration Via the citric acid cycle Fermentation pyruvate fermentation Respiration products lactate ethanol and CO2 pyruvate is oxidized completely to C02 de nes the citric acid cycle Carbon Flow in Citric Acid Cycle 391 Production of Acetyl CoA is pivotal Supplies the energy fOr formation of oxalacetate from which citric acid is derived Further reactions drive synthesis of CO2 Three molecules of C02 are released for each pyruvate molecule oxidized electrons from NADH are transferred to 02 or other terminal acceptors Via the electron transport chain Citric acid cycle generates more energy than does fermentation Presence of an electron acceptor like 02 allows fOr complete oxidation of glucose to CO2 and yields more energy during the process NAD CoA Pyruvate three carbons NADH 002 Key C2 d AcetyCoA C Cltrlc 2101 C4 30A 5 1 Ce cyC C M 39 Oxalacetate2 Citrate NADH NAD Aconitatea Carbon Warez lsocitrates39 ow NADltP Fumaratez FADH j C02 FAD SUCCinatez39 u Ketoglutarat39ez39 39 39 NADPH SuccinyI Co GOA NAD GOA GDP Pi N ADH 00 a GTP 2 Energetics Balance Sheet tor Aerobic Respiration 1 Glycolysis Glucose 21IAD 2 ATP 2 Pymvate 4 ATP 4 ADP to CAC to Complex I a SubstrateleveH phosphorylation 2ADPPi2ATPgtlt2 3ATP b Oxidative phosphoryHatiom 1 is 6 ATP 2 CM Pyruvate39 4NAD GDP FAD r 3 C02 quotT Flint1 GTP to Complex I to Complein a Substratelevel phosphorylatlon 391 GDP P g1W1 GTP 1 GTP 1 ADP 4 ATP 1 GDP 3 Oxiciative phasphory ation 12 ATP 391 FAiDH 2 ATP E 15 ATP x2 3 Sum Glycolysis plus MI gt 38 ATP per glucose Figure 522h Brnck Biolngy at Micmarganisms 11 fie LE lit6 Pearsnm Prentice Hall Inc The Citric Acid Cycle Is Key For Biosynthesis and Is Not Just For Energy Generation Catabolic alternatives to fermentation and respiration Anaerobic respiration Chernolithotrophy citric acid cycle Organic compound 37 Electron transport ATP W Proton mOt39Ve f rce 33 Biosynthesis E g E m Carbonflow 02 So N03 5042 Organic e Aerobic nitrate sulfate I electron acceptors Anaerobic respiration doesnotuseoxygen Chemoorgunoirophic metabolism Figure 523a Brock Biology of Microorganisms 112 9 1006 Pearson Prentice Hall Inc hvdrooen sulfide HZS Fe hvdrooen Gas ammonia NH3 C i C C electron donors Inorganic compound C02 39 ll Electron transport Carbon ATP MProton motive force flow 39 i l M 2 5 02 N03quot 5042 Biosynthesis stable oxygen Chemoliiholrophic metabolism Figure 523b Brock Biology of Microorganisms 1 We 9 2006 Pearson Prentice Hall Inc Typically aerobic respiration electron transport chain forms PMF uses an inorganic source of carbon for biosynthesis Pholoheterolrophy Light Electron transport Organic compound Carbon flow Proton motive force Biosynthesis photophos phorylation gt ATP Pholotrophit metabolism Figure 523 Brock Biology of Mkroorganisms 11e 2006 Pearson Prentice Hall ln PholouIIlolrophy inorganic carbon source co2 Carbon flow Biosynthesis Respiration always lead to generation PROTON MOTIVE FORCE to generate ATP phototrophy chemorganotrophy ehemolithotrophy Microbial Growth Bacterial Cells Divide by Binary Fission reproduction of a cell by division into two approximately equal parts Major Steps in Binary Fission VDNA amp DNA replication lt i y gt Signgation amp Septum formation Completion of septum with formation of distinct walls 7 if 1 callseparation Post Binary Cell Division End up with two cells each with a new and an old pole 3 Fimbriae M Protei n 39 Septum Ar I Chromosome Homologous to eukaryotic tubulin Hiighly conserved mitochondria ch oroplasts 39 Assembles into a ring around the cell FtsZ molecules polymerize Enzymatic activity hydrogze guanosme triphosphate TP Assembly of FtsZ at the divisome With other proteins that participate in cytokinesis Se ressembw m nsz am can ml pulm mm 1 nm naugmerceus laymen 5 1 mm components dlsverse E E E E i39 DAPI FtsZ STAIN FtsZ ring Divisome Cytoplasmic PYOtelnS membrane Division piane quot T den Blaauwen 8 Tnne Nanninga Univ of Amsterdam a in a N I Outer membrane 239 Peptidoglycan Cytnpasmic membrane ivisome Ftsz ring complex Cytnplasmlc membrane 39 A Dmsmn plane a 3quot m m E c g m 2 a I I E n E 33 am ME E42 mm 52 2 Ii Cnpyrighu m 2009 F39Earsnn Educatinn Inn publishing 5 Pearsnn Benjamin Cummings Minute 3 k Cell wall 1 7 V In I r V 39 3 a 1 u 7 V 39 V 2 quot1 I hi4 a l 1 2 I membrane 7 7 7 i 7 7 7 my h Jgga v 1 M Cytoplasmic l J N ucleoid MinE Divisome complex o39 Z g Jup r39rf Hf 339 ea gar jiIJrJafzi3r m 1513 s 39ensudr m vajurr39lmii PquotL1 IUCII JF39 rltWEr I1 blll1393 F d quot IEI f Inquot 1y FtsZ mutant showing lamentation Gabe McCool and Maura Cannon University of Massachusem Amherst FtsZ is dynamic assembly and disassembly of the cytokinetic ring allows progression through cell division cycle Peptidoglycan synthesis critical for productive cell division FtsZ ring 039 e W Wall bands I Growth zones I 4 I r I V I V 4 I HOW do you get the N acetyl lucosamineNacetylmuramic acidpentapeptide peptidoglycan 1 EH3 C3H3 I3H3 Bactoprenol undecaprenolphosphate C55 I Growing point of Peptldoglycan 0er wall Cytoplasmic membrane Out mm 6 up my mg Bactoprenol Transpeptidation rxn crosslinkage of two peptidoglycan chains l 1 Target of penicillin f gt ENERGY Mg L Ala DGlu DAP DAla DAla RELEASED Transpeptidation DAP DGIULAIa39 D Ala m M LAla DGlu DAP D Ala DAP DGlu LAIa 7 DAla K9 5 What is bacterial growth What does that term mean Population Growth Doubling time same as Growth rate calculate as Number m ns 3 rithmetc hr Number f cells Igarithmc Number fGEHE Which culture has a faster growth rate A or B Klett Units 101 2D U n 145quot T215 7370 35 Timehr Phases of Bacterial Growth Lag Log Stationary gelath Bacterial Growth Curve E ElmNth phases HEB Lag Exponential Statmnaw 90 a E J 1113 quotquotquotquotquotquot I quot39rargia y mg 5 30 npticat density j E 39 050 i m f 3 TD l E 4325 250 B D 3 50 e H Time Measurements of Bacterial Growth QDirect microscopic count OViability serial dilutions OTurbidity optical density spectrophotometer
Are you sure you want to buy this material for
You're already Subscribed!
Looks like you've already subscribed to StudySoup, you won't need to purchase another subscription to get this material. To access this material simply click 'View Full Document'