⒈ Amylase Experiment

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Amylase Experiment



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The Digestion of Starch by the Enzyme Amylase

When the nutrients required for the bacteria to grow are abundant, they exhibit metabolic activity. These organisms can produce antibiotics during sporulation. Examples of the antibiotics that Bacillus subtilis can produce include are polymyxin, difficidin, subtilin, and mycobacillin. Many of the Bacillus microbes can degrade polymers such as protein, starch, and pectin, therefore, they are thought to be an important contributor to the carbon and nitrogen cycles.

When they cause contamination, they may result in decomposition. Quite a few of the Bacillus organisms are primarily responsible for the spoilage of food Todar. Bacillus subtilis supports plant browth. As a member of Bacillus , this bacterium often plays a role in replenishing soil nutrients by supplying the terrestrial carbon cycle and the nitrogen cycle. Bacillus subtilis bacteria form rough biofilms, which are dense organism communities, at the air and water interface. Bacillus subtilis biofilms are beneficial. They allow for the control of plant pathogen infections. The plant benefits because B. Preemptive colonization prevents other pathogens from infecting the plant because B. The biofilm communities form a mutualistic interaction with plant rhizome systems.

Bacillus subtilis biofilms found in the rhizosphere of plants promote growth and serve as a biocontroller. In this sense, B. Bacillus subtilis strains can act as biofungicides for benefiting agricultural crops and antibacterial agents. Bacillus subtilis also reduces mild steel corrosion Morikawa Bacillus subtilis bacteria are non-pathogenic. They can contaminate food, however, they seldom result in food poisoning. They are used on plants as a fungicide. They are also used on agricultural seeds, such as vegetable and soybean seeds, as a fungicide. The bacteria, colonized on root systems, compete with disease causing fungal organisms. Some strains of Bacillus subtilis cause rots in potatoes.

It grows in food that is non-acidic, and can cause ropiness in bread that is spoiled Todar. Some strains related to Bacillus subtilis are capable of producing toxins for insects. Those strains can also be used for protecting crops as well. Some Bacillus species can cause food poisoning, such as Bacillus cereus and Bacillus licheniformis. Bacillus cereus can result in two different kinds of intoxications. It can either cause nausea, vomiting, and abdominal cramps for hours, or diarrhea and abdominal cramps for hours.

Some Bacillus organisms can cause more severe illnesses. Bacillus anthracis , for example, causes Anthrax. It was the first bacterial organism that was known to cause disease in humans. Bacillus anthracis spores can survive for very long periods of time. Anthrax is very rare in humans, however it is more common in animals. Bacillus organisms, isolated by soil sprinkle technique, are responsible for producing antibiotics. The most antibiotic activity was seen in Bacillus subtilis MH The most optimal activity occurs at a temperature of 37 degrees Celsius and a basic pH of 8. Glycerol is the optimal carbon source and L-glutamic acid is the optimal source of nitrogen.

The antibiotic bacitracin was determined to be affective on Gram-positive bacteria only Jamil Other antibiotics that Bacillus subtilis form are polymyxin, difficidin, subtilin, and mycobacillin. Polymyxin is affective against Gram-negative bacteria, whereas difficidin has a broader spectrum Todar. Bacillus subtilis bacteria secrete enzymes, "such as amylase, protease, pullulanase, chitinase, xylanase, lipase, among others. There are many research studies that are currently being done on Bacillus subtilis.

One recent research project focuses on the resistance of Bacillus subtilis spores to heat, radiation, and chemicals. It has been known that spores can survive hundreds, even millions, of years in a dormant state. The study investigated the important factors that contribute to spore resistance. The researcher found that the bacteria's coats were a major factor because the coat provides a barrier for the organism against toxic agents, ultraviolet radiation, and lytic enzymes. The inner membrane was also found to be important, due to its low permeability against toxic agents. DNA repair was also determined to be crucial, since it can control DNA damage due to radiation, heat, and toxins.

Bacillus subtilis spores are also resistant to wet heat, primarily by the core's low water content. The lower the water content of the core is, the more resistant the spore is to wet heat. This research study is important in that it can lead to future studies on how the Bacillus subtilis spores in food and medical products can be killed effectively. Learning about the spores resistance gives us a better understanding of which methods may or may not be useful in killing the spores Setlow Another current research study provides evidence that the SpoIIIE DNA translocase is required for Bacillus subtilis forespore chromosome translocation across the septum and membrane fusion during sporulation.

They found that one mutant undergoes the translocation of DNA, but does not undergo membrane fusion normally after the engulfment. They discovered that the septum stays open in this mutation. When the sporulation septum is open, the cytoplasm is permitted to be exchanged between the daughter cells. This implies that the membrane does not fuse properly after engulfment and cytokinesis. The researchers proposed "that SpoIIIE catalyses these topologically opposite fusion events by assembling or disassembling a proteinaceous fusion pore" Liu The study demonstrated that SpoIIIE first participates in allowing a barrier of diffusion for the translocation of DNA, and then participates in the fusion of the membrane.

A third current research project investigates Bacillus subtilis fermented soybean meal and its effects on enzymes in the gastrointestinal tract and intestinal morphology of piglets. The piglets were randomly given either soybean meal or fermented soybean meal. After the experiment was completed, the six piglets from each of the two treatment groups were sacrificed. The contents of the small intestine were collected, and the tissue was sampled at varying locations. The researchers found, using light microscopy, that the piglets in the treatment group that were fed fermented soybean meal had significantly taller villi at the varying locations, and had a significantly lower duodenal crypt depth in comparison to the piglets in the treatment group that were fed soybean meal.

They also showed a significant increase in duodenal and jejunal protease and trypsin activities and a decrease in pancreatic trypsin activity. The findings obtained from this research demonstrate that fermented soybean meal improves the morphology of the intestine as well as the activities of digestive enzymes Feng Journal of Bacteriology.

Digestive Diseases and Sciences. Bacillus : Cellular and Molecular Biology. Caister Academic press. ISBN Pak J Pharm Sci. Dutton, K. Mol Microbiol February;59 4 J Bacteriol. Journal of Bioscience and Bioengineering. Annual Review of Microbiology. Abanes-De Mello, K. Schaechter, M. Ingraham, F. Journal of Applied Microbiology. Edited by a student of M Glogowski at Loyola University.

From MicrobeWiki, the student-edited microbiology resource. Jump to: navigation , search. This is a curated page. Report corrections to Microbewiki. A Microbial Biorealm page on the genus Bacillus subtilis Contents. Bacillus subtilis colony on TSA exhibiting raised, dull, wrinkled characteristics. Glogowski with permission. Plot a graph of rate of reaction against pH. A similar experiment can be carried out to investigate the effect of temperature on amylase activity. Set up a series of test tubes in the same way and maintain these at different temperatures using a water bath — either electrical or a heated beaker of water.

Depending on the chemical reaction under investigation, you might monitor the reaction in a different way. If investigating the effect of temperature on the breakdown of lipid by lipase , for instance, you could monitor pH change — lipids are broken down into fatty acids and glycerol. As the reaction proceeded, the release of fatty acids would mean that the pH would decrease.

Required practical activity Investigate the effect of pH on the rate of reaction of amylase enzyme Aim To determine the rate of the amylase activity at different pHs. Method You will investigate the breakdown of starch by amylase at different pHs. When the starch solution is added: start timing immediately remove a sample immediately, and test it with iodine solution sample the starch-amylase mixture continuously, for example every 10 seconds This is how you might set up the experiment: For each pH investigated, record the time taken for the disappearance of starch, ie when the iodine solution in the spotting tile remains orange-brown. Hazards Wear safety goggles. Amylase solution may cause allergic reactions. Iodine solution is irritant. Avoid contact with skin and eyes.

Example results pH Time taken for the disappearance of starch in s 5 6 40 7 30 8 50 9 The time taken for the disappearance of starch is not the rate of reaction.

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