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Date Created: 08/17/15
MicrobiologyJune 132015 Exam 2 Chapter 5 Microbial Metabolism Metabolism Two types Catabolism provides energy and building block for anabolism Anabolism energy and building of more complex molecule from simple molecules Both are coupled together Collision Theory Activation energy energy needed to disrupt electronic con guration Can occur with or without enzymes Enzymes will lower the activation energy Enzyme are bioogica catalyst Apo enzymeprotein part 2 parts of an enzyme Cofactor none protein part Holoenzyme Apo enzyme cofactor enzymes binds to the substrate at the activate site The substrate is converted the product The enzymes will remain unchanged 0 Factor that in uence Enzyme Activity Both the temperature and the PH will denature protein As the temperature and increases at a certain time the enzyme activity will decrease Substrate concentration With increasing concentration the enzyme activity will increase until all of the activate site is all lled up It will then become saturated Competitive inhibitor the competitor has a similar structure as thee substrate and they both compete for the activate site Noncompetitive inhibition the competitor will bind to a site other than the active site EX 0 Feedback inhibition the enzyme will inhibit the activity of the rst enzyme in the pathway OxidationReduction Reaction Oxidation removal of an electron Reductiongain of an electron Redox reaction the coupling of an oxidation and reduction reaction ATP is generated by the Phosphorylation of ADP SubstrateLevel Phosphorylation ADP ATP by just adding on an electron Energy from the transfer of a high energy PO4 to ADP generated ATP Oxidative Phosphorylation ATP made from electron transport chain Photophosphorylation ATP made from photosynthesis Causes the chlorophyll to give up electron The Breakdown of Carbohydrates to release energy Glycolysis Oxidation of glucose to pyruvate Products ATP and NADH In gycoysis start with glucose Glucose is a 6 carbon compound Substrate 2 ATPs are used 6 carbon compounds are split into two 3 carbon compound After the split 2 NADH and 4 ATP are made Glycolysis nal product 2 Pyruvate Net ATP 2 Before the Krebs cycle the pyruvate cycle The pyruvate acid is decarboxylated and converted to acetyl COA 2 Pyruvate 2 Krebs cycle Krebs cycle 3 NADH 2 6 1 FADH 2 1 ATP 2 T ETC EukHappens in the lntermembrane For every NADH 3 ATP FADH 2 ATP The ATP that is made from the ETC is an example of the oxidation phosphorylation Glycolysis Net 4 ATP 2ATPs 2 2NADHx3ATPsin ET 6 2 Pyruvate Acetyl CoA 2 NADH X 3 ATP 6 ATP Kreb 3 NADH 3X3 ATP9 1 FADH 2X12 1 ATP 1X1 12 ATP from the krebs cycle 36 ATP In Euk 24 14 36 total AATP 38 ATP in Pro Aerobic respiration nal electron acceptor is oxygen Anaerobic respiration nal electron acceptor is an inorganic compound other than oxygen Fermentation the nal electron acceptor is an organic compound Lipid Catabolism Simple molecule of lipids1 one molecule glycerol and 3 molecule of FAs Will enter during the 3 carbon split 1 NADH 3 ATP 3 Net ATP 2 ATP 2 1 PYRUVATE Aceytl C0a 3 Biochemical testing Photosynthesis Light reaction Make ATP Dark reaction Fix C02 living things must have a source of energy and a source of carbon Source of energy is lightphoto Source of energy is chemical or an organic compound chemo Source of carbon is C02 autotroph Source is an organic compound chemoautotroph Amphibolic Pathways Have both catabolic and anabolic pathways Lab acid fast stain and read sporostain procedure July 15 2015 Next Exam Tuesday Chapter 56 7 Microbial Growth Increase in the number of cell not the size Factors that affect growth Temperature 1 Min Growth Temp 2 Op Growth Temp 3 Max Growth tem Psytrophiles Cold loving Min 10 Pathogenic Bacteria are mesophies Min 10 Max50 Opt35 Psychotrophs Causes food spoilage in the refrigerator PH 0 Most bacteria will grow between PH 6575 Molds and yeast grow between PH 5 and 6 Acidophiles grow in acidic environments Osmotic Pressure Hypertonic environments or an increase in salt or sugar cause Pasmoysis Extreme or obligate haophies require high osmatic pressure Halophiles require high salt lsotonic solution if solute concentration outside 085 the solute concentration inside is going to be 085 Nitrogen Sulfur and Phosphorus requirement Requirement for oxygen Obligate aerobes require oxygen for growth Facultative anaerobes Can grow in the presence of oxygen or absence Obligate anaerobes are killed in the presence of oxygen Why are anaerobes killed in the presence of oxygen In the presence of oxygen the molecular oxygen is converted to a superoxide radical Aerobes will take two superoxide radical and will convert it into hydrogen peroxide In order to convert superoxide radicals you need the enzyme superoxide dismutase 1 The Hydrogen Peroxide is toxic to cell but not as toxic as the super oxide 2 The hydrogen peroxide can be broken by a catalase or a peroxidase Aerobes have the superoxide dismutase catalase and perodidase they have 1 or more for disper The will have a superoxide dismutase Anaerobes do not have these enzyme so you will have the buildup for the superoxide which will be toxic to the cell This can killed them How to grow anaerobes You can put chemicals to kill the oxygen Growth Chamber Anything that will increase the c02 leel will decrease 02 level Capnophiles Bio lmA group of bacteria that migrate together Advnatages Bio ms share nutrients Shelter each other from harm Pseudomonas aeruginosa bio lm they produce a bluegreen color They are an opportunistic Big problem in surgical and burn units They can cause an infection Agar Complex polysaccharide Used as solidifying agent for culture media in petri plates slants and deeps Generally not metabolized by micr Microbes Makes it east distinguish colonies of different microbes Autoimmune disease Destroying your own cells Selective Media Suppress unwanted microbes and encourage the desired microbes Ex Mannitol plate only staph Aureus will grow on the mannitol Obtaining Pure Culture Streak plate method used to isolate a pure method Fg 611 When burning your decreasing the number of cells when you go from one area to another How to preserve bacteria by Deepfreezing and freeze drying Most cells are going to divide by binary ssion Growth curve important 1 2 3 Chapter 8 Sespsis Lag culture is overcoming the prime media preparing for population growth but in increase in population Log PhaseThe number of cells diving is greater than the number of cells dying Stationary phase The number of cells dividing and dying is equal Death PhasePopulation is decreasing at a logarithmic rate Turbidity Cloudiness Bacterial growth causes cloudiness If you have growth you will have a decrease in the amount of light Measuring Microbial Growth Direct method Indirect Method Refers to microbial contamination AsepsisIs the absence of a signi cant contamination Sterilization removal of all microbial life Commercial Sterilization Killing C Botulinum endospores Disinfection removing of pathogen Antisepsis Removing pathogens from living tissues DegermingRemoving microbes from a limiting area Ex Giving a shot Sanitization Lowering microbial counts on eating utensils BiocidegermicideKilling microbes Bacteriostatic Inhibiting not killing microbes Effectiveness of treatment Depends on 1 Number of microbes 2 Environment 3 Time of exposure 4 Microbial Characteristics Actions of Microbial control agents 1 Alteration of membrane permeability 2 Damage proteins 3 Damage nucleic acids Moist Het Sterilization Most Heat denatures proteins Autoclave high temp and high pressure Pasteurization High temp for a short period of time It kills pathogens and organisms 91 Dry Heat Sterilization Kils by oxidation Flaming Dry heat ncrineration 99 F Filtration Inhibit microbial growth Physical methods Low tempinhiibits miciall growth Re rgerationdeepfreezing Lyophilization High temp denatures protein Desiccationprevents metabolism drying Ionizing radiation Xrays Gamma rays electrons beams Ionizes water to release OH Damages DNA Nonionizing Radiation UV Principles of Disinfection 1 concentration id disinfectant 2 Organic matter 3 PH 4 Time Figure 76 Dilution Test The greater the zone of imbibition the more effective the chemical or antibiotic is More zone of inhibition The chemical will diffuse out Which is more effective Undiluted g 71 PG 202 will give speci c mechanism of actions Know that alcohol 7890 is more effective than 100 because dilution with water is require P ons Infectious protiens 07202015 Gene segment of DNA that encodes a functional product usually a protein Replicate Replicate all of thee dna Transripetion codes for genes Genotypethe genes of an organism genetic make up Phenotype the physical expression Genetic information ow DNA RNA Transcription RNA Protein Translation Plamid extrachromosomal DNA that can carry genes DNA Polymer of nucleotides adenine thymine cytosine and guanine Double helix associated with proteins Backbone is a deoxyribosephosphateFree hydroxy Strands are held together by hydrogen bons between AT and CG Strands are antiparell 539 prime ends Free phosphate 339 prime free hydroxyl The place where thee DNA unwinds to start replication is the replication fork Main enzyme in DNA replication DNA polymerase The DNA polymerase only goes in one direction 5393 direction Made in a continuous leading strand Discontinuous Lagging strand Okazaki fragments Transpcription makes proteins 3 types RNA Mrna Trna rRNA Transcription begins at the promoter Promoter where the RNA polyermase will made ana polymerase starts transcription Transcription will stop when it reaches the terminator In eukaryotic cells introns exons More gentric material then needed ntrons are removed exons come together which makes the messenger RNA On the messenger RNA Codon three nucleotides Start codonAUG Translation ends when you reach a nonsense codon UAAAUAGUGA 64 Codons for 24 amino acids tRNA cares the anti codon the trna will bring in the amino acid the anti codon is complementary to thee codon Trna anti codon Mrna codon Constitive genes are expressed at a xed rate Other genes are expressed only as needed Inducible genes are turned on Repressible genes are turned of o A group of similar gene under one control operon Control region a promoter and an operator The promoter is the place where the RNA polymerase binds to initiation transcription The I gene codes for a repressor The repressor can be active or inactive If the repressor is active it will bind to the operator It will prevent the RNA polymerase from binding to the promoter Mutation change in gentric material Base subisition point mutation change n in one base Misense mutation Base subitution results in the change in amino acid Nonsense mutation base substitution results in a nonsense codon Stop codon does not code for any amino acid Frameshift mutation lnsert a base or delete a base Auxotrophic adding something in the media to get a bacteria to grow Ames test The rst amino acid is Met Horizontal gene expression Horizontal gene transfer gene transfer between similar Vertical gene transferreproduction Methods of gene exchange Transformation uptake if unattached DNA Congugation cell to cell conguation F factor plasmid Plasmid extra Chromosome nothing for the survival of cells When you make FF news cells F HFR high frequency of recombination Mutiple replicating parts Transduction method of change exchange where virus is used Bacteriophagevirus that affects bacteria Jumping gene Naked DNA F methlyine Translation Vector Restriction enzymes Cuts DNA in speci c places How does a cell chop up foreign DNA and not its own DNA The cell39s DNA is methylated Restriction enzymes Cut speci c DNA Vectors Carry new DNA to desired ce Shuttle vectors can exists in several different species Plasmids and viruses are used as vectors Polymerase chain Reaction To make multiple copies of piece of DNA enzymatically Used to Cone DNA for recombination Amplify DNA to detectable levels In each cycle Inserting Foreign DNA into cells Separates DNA fragments of different sizes DNA moves toward the positivelv charged end of the gel Transformation exchanging naked DNA protopast Fusion Obtaining DNA Complementary DNA is made from an enzyme from mRNA by reverse transcriptase Mrna uses reverse transcriptase cDNA minus the introns Methione rst protein related to translation How to detect thee gene of interest Coony hybridization Florescence abeed probe It will hybridized with the same sequence DNA nger print Isolate the DNA with the restriction enzyme DNA to DNA Hybridization Hybridize probes to DNA fragments that have been separated by gel electrophoresis RNA to DNA Hyberization Hybridized pair Hybridize probes to RNA fragments that has been separated by gel electrophoresisre All of these are ways to determine the gene of interest GMOs must be safe for consumption and for the environment Steps of recominiamt adna Isoate Dignestion with restriction enzyme have a vector Seection process Exam 3 89 The following questions refer to the mRNA molecule seen below 539 AUGGGAAU UAGAUCCUG UGCAUAAGACCAGAG UCAA 339 1 What would be the rst three amino acids synthesized upon translation of the mRNA 2 points MetGIy Iso 1 How long how many amino acids would the protein produced from this mRNA be 2 points Seven note the UAA stop codon 1 What is the name of the part of the tRNA which would bind to the triplet AUU in the mRNA above 1 point Anticodon 1 Would a mutation which caused the triplet GGA to be changed to GGU be a serious problem to a microorganism in most cases Why or why not 2 points No GGA and GGU both code for the amino acid glycine Answer the next three questions assuming that because of exposure to UV radiation a single quotAquot was introduced into the fourth position of the mRNA above so that the sequence now reads 539 AUGAGGAAUUAGAUCCUGUGCAUA AGACCAGAGUCAA 339 1 What would be the rst three amino acids produced 1 point MetArgAsp 1 What type of mutation could this be characterized as could it be characterized as more than one type Why What is the most serious aspect of this type of mutation 2 points This is both an insertion and a frameshift mutation The most serious aspect is the fact that the reading frame of the ribosome is changed resulting in a change in every amino acid in the protein after the point of the mutation Chapter 10 Taxonomy The scienctic classifying organisms The 3 domians of classi cation Prokaryotes Eukaryotes and Archea Lack peptidoglycan in cell walls singlecelled prokaryotes thrive in extreme environmental conditions salty hot acidic extremophiles limited to harsh conditions because they can39t compete with Bacteria more closely related to Domain Eukarya Both the bacteria and the archea are both Prokaryote but the Archea will not have peptidoglycan in their cell wall FossHs Honors the person who discovered may give something about the disease Domian Kindon Phylum Class Order Family Genus Species Population of cells derived from single cells Eukaryotic species A group of closely relate organisms that breed among themselves Viral Species population virus with similar characteristics that occupies a particular ecological niche Classi cation Placing organisms in a group of related species List of characteristics of know organism Flow Cytometry Gene cs DNA base composition Guanine cytosine moles diagram that helps you deliver species Chapter 11 obligate Transmitted by Arthropodborne spotted fevers Gram positive bacteria Legionellales Found in streams warm water ipes coling towers L pneumophilla Home work What is the growth requirement for legionella Vibrionales Found in Coastal Water Gatriontristis Enterobaterilais Gram Negative Puruple They inhibit the intestinal tract 8 Ebterobacter Erwinia Esche cha Klensiella Proteus Salmonella Serriatia Shigella Yersinia Pasteurellales Bacteria found in the blood Chemautotrphic Endospore Producing Obligate Anaerobes Endospre forming gram positive Cocci Clustered schains and gram positive Mycoplasma No Cell Walls Mycology The study of Fungi Examples Molds Opportunistic bacteria Lichens combination between and algae and a fungus They live together and bene t each other Economic advantages 3 methods of movement Finger like projection Flagella Cillia Kingdom Animalia Phylum Plathelminthes Flatworms Nemaota Roundworms Viruses that effect bacteria Bacteriophage Bacteriophages form plaques clear area causes lysis When normal cells come in contact with each they stop growmonoayer Transformed cells do not form monolayer they will continue to grow when they do not come in contact with eac other Virsus inti cation Cytopathogenic effect Pattern of ysis patterns Lytic cycle2 Multiplication of Animal viruses Attachment Virus attach to cell membrane PenetrationBy endocytosis or fusion Attaches the DNA to the host cell animal cells Prophage Inserted DNA Unclotting vy viral or host Lysogenic cycle Incorporation of the genetic material from the host cell Can be a RNA or a DNA You cannot incorporate an RNA virus into a DNA Cancers Associated with virsus Activated oncogenes transform normal cells into cancerous cells transformed normal cell into cancerous Cancers associated with virus is an oncogenes Transformed cell will increase growth loss of contct inhibition tumo speci c transplant antigens T antigens The genetic material of incogenic virsus becomes integrated into the host cell DNA Oncogenic RN virus Needs a reverse transcriptase Virsus remain a symphamtic for a long period of time Outbreaks with system Herpes Chicken pox Prion infectious protein Inhertited by transmissle by ingestion transplant and surgical instruments Normal PrPC PrPSC39Not normal Immunology Final Microbiota Normal Microbiota protects the host cells by occupy niches that pathogens might occupy Producing acids and bacterium39s ProbioticsLive Microbes applied to or ingested into the boy intended to exert a bene cial effect Epidemiology The study of disease Endemic Infections graph Latent Infectious Disease with a Factors that contribute to a pathogens Invasiveness Age Chemeeotherpy Fatique Lifestye Climate and Weather Short Urethra in Feemales nherited traits such as the sickle cell gene Mechanical Transmission Arthropods carries Pathogen on feet Biological Transmission Pathogen reproduces in vector Prevented by use if gloves Face shield using aseptic procedures Patient are susceptible to Nosocomial infections because they already have weak immune system protein Are from gram negative Are part of the outer membrane From the lipid oirtions LPS Only released when the cell disease or divides This helps stimulate Pathogens Proteins produced in the pathogenic path way What is complement A second line of defense Set of blood plasma proteins that act as chemotactic attractants trigger in ammation and fever and ultimately destroy foreign cells Complement A group of serum protein that is in your body that is activated in a cascade protein Classical pathway Activated when the Antibody is binding to the Antibody Alternative pathwayActivated by C3 and Microbes The main line of defense is for viral infection is Interferon Interferon are proteins are released from the infected cell That keep the uninfected cell from becoming infected Humoral response be cells B cells make antibodies B cells will differinante in the bone marrow Cellular immune response Make T cells will differiante in the thymus An antibody is a protein is make by your body in response to the antigen A substance that cause the body to produce speci c antibodies or sensitize T cells The antibody is a Globular proteins are called immunoglobins Structure of Antibody include 2 heavy chain and 2 light chain They are polypeptide chain On the heavy and the light there is a V region and constant region Amino Acids are more variable in the V region In the C region they are more constant The place where the Antibody bind to the Antigen is at the variable region The lg is the largest part of the antibody It is the Ig is the only Antibody that crosses the Placenta lgM it is a pentomerlt is the rst Antibody to be produced in an immune response lgA Dimer found in external mucosal secretion lg DMonomer initiate an Immune Response lgE Monomer responsible for allergic regions and lysis of parasitic worms 0 If you have a disease where the toxin is causing the disease You reat it would a toxoid Toxoid lnactiviatited toxin used in vaccines Immortal cancerous B cell fused with an antibodyproducing Normal B cell Produces monoclonal antibodies Monoclonal Antibodies are used to treat a lot of diease You get Antibody speci c to that B cell Westing Blotting To test Antigen Anytime you have the same antigen as Antibody agglutination Hemeaggultination Neutralization test Used to etect disease Compliment Plate you get from the pharmcutical company To test disease direct The antibody is bond to the plate you add the antigen If the antigen is bound you add to the antibody direct Elisa The antigen is bound ELISA stands for EnzymeLinked ImmunoSorbancy Assay DD ELISA is used for test for the presence of against speci c Antibodies Antigens DD Examples of ELISA testing 3 Testings for HIV pregnancy illegal narcotics DE The ELISA is based on the bond that occurs between an antigen and it39s corresponding antibodiy ultratight DD Theory behind ELISA in response to a disease the host will the host will produce antibodies that bind to the antigen associated with the disease DD To perform an ELISA test you combine 3 1 the antigen usually puri ed form 2 the antibodies from the patients serum sample 3 an indicator that allows you to visually determine if the antibodies bonded to the antigens usually a synthetic combo of secondary antibody and an enzyme DD Controls are used to safeguard against what 2 things poor technique and crosscontamination is when an ELISA returns to a positive result but the patient doesn39t have the disease False positive DD is when an ELISA returns a negative result but the patient actually has the disease False negative DD Positive and Negative controls are done at the same time that the actual sample is being tested to ensure what that the protocol worked DD Performing an ELISA 1 add pure disease ANTIGEN to wells of plastic microplate strip 2 add the PATIENTS SAMPLE and the and control samples to the appropriate wells 3 Add SECONDARY ANTIBODY to the wells this attaches to enzyme HRP 4 Add SUBSTRATE that is acted upon by HRP e look at Side 192 Y Blood Type A A anigrn Anti Body BB antigen Anti Body AB both No Antibody universal recipent 0 no antigen have both A and be Antobody Universal donor You cannot Hemlytic sdisease Rh factor Problems with pregnancy Rh male with an Rh and an Rh fetus a drug will be given At the birth of the second birth mother will make antibody to the Rh The IgG will go to the placenta Autommune disease Lost of cell tolerance Your destroying your own self Organ transplant Math the genes from the major histone compatibility complex
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