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Microbiology Lab Final

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Biological Sciences
BIOL 2051

Microbiology Lab FinalCatalase productionEnzymes catalyze reactions of metabolismAnd enzyme can generally only react with one substrate and can catalyze one reactionExtracellular enzymes are enzymes that act outside of the cell in which they are produced Endoenzymes act inside the cell to degrade molecules to simpler molecules and release energyEnzymes that are produced only when the substrate is available is called inducible or adaptive enzymes Enzymes that are essential at all times are called constitutive enzymesMicrobiologist often investigate and organisms metabolic capabilitiesThis type of assessment tells how the organism functions in an environment and is a study of the physiology of that organismCatalase breaks down toxic hydrogen peroxide into waterUsed to distinguish between organisms especially streptococcus and staphylococcusStreptococcus Lactococcus and Enterococcus are Catalase negative anaerobic or fermentative Stapylococcus and Micrococcus and Catalase positive aerobic Drop H2O2 add organism observe for bubblesStaphylococcus epidermididis is positiveStreptococcus mutans is negativeOxidase ProductionOxidase enzymes are made by organisms that produce energy using aerobic respirationAn enzyme called cytochrome oxidase catalyzes the oxidation of cytochrome and the subsequent reduction of molecular oxygen to either H2O H2O2Oxidase tests are used to differentiate medically relevant generaoVibrio and Pseudomonas positive from Enteric bacteria like Escherichia and Salmonella negativeoNeisseria and Moraxella positive from Acinebacter negativeTransfer 200 microliters of oxidase reagent and watch for bluepurple happens fastPseudomonas putida is positiveE coli is negativeHydrolysis of Polysaccharides Carbohydrates CH2OnMonosaccharides ex glucose single molecule of a sugarPolysaccharide ex cellulose starch each are polymers of glucoseAmylase enzyme that hydrolyzes starch into smaller subunits glucose to be transported into a celloMicrobially produced amylases have commercial and industrial usesfood fermentation laundry detergent brewing distilling textiles paper etcoFlood SA plates with organisms with iodine Amylase production will be evident by a clear area zone of hydrolysis around and under the amylase producing organism Iodine reacts with starch to produce a brownblue coloroBacillus subtilis is positive E coli is negativeCellulase is an enzyme that hydrolyzes cellulose intoto be transported into the cell for glucosemetabolismoCellulose is a polymer of glucose but unlike starch it has different glycosidic bondsoCellulose is found in eukaryotic cell wallsoCellulose decomposition is part of the natural cycling of carbonGroup A 4 pieces of paper on dubos agar plateLook for growth on paperGroup B paper in test tubesLook for growth and degradation of paper oCytophaga hutchinsonii is positive for cellulose productionThe hydrolysis of proteinsProteins are amino acids joined by peptide bondsPeptide is a short chain of amino acidsProteases are enzymes that hydrolyze proteins into peptides so they can be transported into the cellCommercial use of proteases laundry detergent stain remover meat tenderizer
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