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Bacillus thuringiensis - “Bt” B. sphaericus LECTURE 14 GRAM POSITIVE BACTERIA (cont.) Both species form a parasporal body - a solid protein crystal next to their spores. These species of bacteria kill insects: • B. thuringiensis - moth larvae (caterpillars) - beetle larvae • B. sphaericus mosquito larvae Genes from Bt have been integrated into several plant genomes to give plants permanent resistance to pests: “Bt” is a popular organic insecticide The Giant Bacterium - Epulopiscium First isolated from a surgeonfish See Perspective 1.1, pg. 14 Assumed to be protozoa because of their size - up to 600 uM DNA sequencing showed however that Epulopiscium is a bacteria, not a protozoan. These bacteria are closely related to Clostridia and Bacillus. In 1997, off the coast of Namibia, an even larger bacterium was discovered (See Perspective 1.1, pg. 14). These bacteria, Thiomargarita namibiensis, are not Gram + bacteria. Also giant Beggiatoa-like Thioploca off the coast of Chile These bacteria use H2S as an energy source and nitrate as an electron acceptor. It’s as big as the head of a fruit fly! Part of the reason that they’re so huge is that as much as 98% of their cell is filled up with a vacuole filled with nitrate. Low GC Gm- cont. Staphylococcaceae - Staphylococcus Table 22.1. Note that the dominant organisms are adapted to dry conditions. • facultatively anaerobic, nonmotile, cocci that form irregular clusters Table 23.1 Fig. 22.1. The skin as a complex landscape for microbes. Note that High GC, Gm+ bacteria dominate the deeper zones of the hair follicle. These organisms produce fatty acids (e.g. propionic acid) by fermenting oils from the sebaceous gland. Low GC Gm- cont. The Nose and upper respiratory tract Staphylococcus aureus and S. epidermidis are the predominant bacteria in the nose. See Table 23.1 20% (much higher % among hospital workers) of humans carry S. aureus, an opportunistic pathogen Fig. 22.3. Boil or pimple formation caused by coagulase-positive Staphylococcus aureus. Lactic Acid Bacteria Fig. 6.23. Fermentations • produce lactic acid as fermentation product !Homolactic fermentation only lactic acid (2 per sugar fermented) !Heterolactic fermentation other things, too (usually 1 lactic acid and 1 acetaldehyde or alcohol) Table 32.1 Lactic acid bacteria are fermentative bacteria that can tolerate O2 but can't use O2 in their metabolism (aerotolerant). Live in rich environments (like your throat or milk) and have lost the ability (through evolutionary time) to synthesize many amino acids and vitamins. Such organisms are referred to as fastidious. • Common genera are: Streptococcus, Enterococcus, Lactococcus, and Lactobacillus. Streptococci (strept = Gr. for twisted) • some are important pathogens (e.g. S. pneumoniae and S. mutans). • very common inhabitants of the human body and foods (will come back to in a few minutes…). • Don’t produce spores Homolactic fermentation (produce 2 lactic acid for every glucose used in fermentation) Fig. 11.3. Streptococcus spp. Streptococcal Diseases • various Streptococcus species We will concentrate on tooth decay (dental caries today (read pgs. 602-604) Later in semester… Strep Throat (Steptococcal Pharyngitis, pgs. 565-569) and S. mutans is the primary cause of dental caries via an plasmid-encoded enzyme dextransucrase which catalyzes the following: n sucrose ----> dextran + n fructose Dextran is a sticky polymer (alpha-1,6 linkages) of glucose molecules (we don’t have the enzyme to cleave that bond). A little bit about pneumonia (pgs. 576-579) Remember S. mutans is also homofermntative so it also converts every fructose from above to 2 lactic acids n sucrose ----> dextran + 2n lactic acid Caries are caused by this acid eating away at the enamel…. Sucrose and S. mutans are needed because S. mutans can’t cause caries without sucrose….. and S. mutans-free animals don’t get cavities even in the presence of sucrose Fig. 24.3. Dental Plaque. Many types of Bacteria in a polysaccharide matrix. Fig. 24.4 Increase in acidity after sugar addition to dental plaque. • Lactobacillus used to make many foods - yogurt, sauerkraut, beer, wine, cheese, sour dough bread… Usually rods (see Fig. 11.4) can live at lower pHs than Streptococcus spp. thus is important in later stages of food fermentations (e.g. in yogurt, sauerkraut). Homo- or heterolactic fermentors. Fig. 11.4. Lactobacillus sp. in yogurt. Note denatured proteins (= curdled milk) Table 32.1 Fig. 30.4 Succession in microbial communities - applications to food spoilage and enhancement….. Succession in foods is often related to pH changes due to lactic acid bacteria…… Start with food spoilage see figure 30.4 making yogurt Streptococcus thermophilus, Lactobacillus bulgaricus (Lactobacillus delbruekii subsp. bulgaricus) Natural successional processes can be manipulated to enhance flavor and preserve food…. 1. Pasteurize milk - kills most of the organisms in the milk (but not S. thermophilus). 2. Inoculate with yogurt 3. Incubate at 45°C (optimum for S. thermophilus). S. thermophilus grows Example of yogurt and cheeses……. And then a bit on wine and vinegar…. and produces lactic acid. 4. Cool when chunky - encourages L. delbrueckii (opt. = 37°C); grow producing lactic acid and aromatic compounds (acetaldehyde) that contribute to the yogurt flavor. Lactobacilli are more acid tolerant than Streptococci. Therefore the above steps enhance a natural successional process. Other organisms are added to yogurt: Lactobacillus acidophilus (acidophilus) Bifidobacterium bifidum (see next lecture) Both can colonize the human colon; have therapeutic value for recovery from diarrhea etc. Cheese…. Curd = coagulated (denatured) milk proteins surrounding fat etc… caused by lactic acid bacteria and rennin = an acid protease from calf stomachs (or fungal acid proteases) Curds are then ripened - 3 main ways to ripen cheese: 1) Original bacteria in the cheese (e.g. Parmesan, cheddar, Gouda, Swiss) 2) Microbes injected inside the cheese (e.g. blue cheese injected with the fungus Penicillium roquefortii) 3) Microbes that grow on the cheese (e.g. brie and Camembert are covered with Penicillium camembertii) Fig. 32.3 Table 32.3 Table 32.2 Let’s look at Soy Sauce (Shoyu) in a little more detail a succession of microbes based on release of different organic constituents over time….. Alcoholic Fermentation - 2 step process PYRUVATE ACETALDEHYDE + CO2 NADH ETHANOL NAD+ Used to make bread, wine, and beer. Fig. 32.04 The difference between white and red wines has mostly to do with how long the must is exposed to the skins (where most of the pigments are) and the occurrence of the malo-lactic fermentation. In both white and red wines yeast (Saccharomyces cerevisiae) are usually added to the wine to perform the ethanol fermentation. But, many red wine grapes are picked before they are completely ripe and as a result there is an excess of malic acid in the grapes (that would be converted to sugar in ripe grapes). After the wine has set for a while and most of the sugars are fermented, certain lactic acid bacteria (e.g. Lactobacillius spp.) carry out a fermentation that converts malic acid to lactic acid and CO2. The Malo-lactic fermentation (cont.) Because malic is a dicarboxilic acid and lactic is a monocarboxilic acid, this reaction can cut the acidity of the wine by up to half - making for a more mellow wine. COOH COOH HCOH ------> HCOH + CH2 CH3 COOH Malic acid Lactic acid CO2 This step is absolutely essential for the production of high quality red wines, especially in Burgundy and Bordeaux, where the growing season is short. Vinegar production from wine……. by Acetobacter aceti (an aerobic alpha Proteobacterium) and related bacteria that oxidize ethanol to acetic acid….. Colonies of Acetobacter aceti (an alpha Proteobacterium) on calcium carbonate agar. Zone of clearing indicates acetic acid production…… One last microbial food…. Edible cyanobacteria Spirulina platensis is used directly as food in parts of Africa and now in Europe… Spirulina platensis is harvested from seasonally dry ponds near Lake Chad… The mats of bacteria are cut into cakes called Dihe. Very high protein content (65%) and very high yield of protein per acre in France (10 tons protein per acre as compared to 0.02 for beef and 0.2 tons per acre for wheat).