Fermentation

Effects of Sugar Revolution – Economic During the seventeenth century the pattern of the Landownership changed from small planters to wealthy individuals and the price of land became extremely high as sugar became more profitable in the Caribbean. Previously tobacco and the other cash crops such as corn were produced by small planters on relatively small plots of land between five and thirty acres. In the year 1645 there were approximately 5000 smallholdings in Barbados that mainly cultivated tobacco, but as the months went by the price of tobacco was gradually falling and ten acres was Just not enough.

The smallholders either moved to another island for a fresh start or returned to England. Consequently the availability of the land increased for larger sugar plantations in Barbados and other Caribbean Islands. Sugar could only be grown on economically large estates so the landholdings increased in size and small landholding were grouped together to make a large estate. They were owned by rich planters, a partnership between two planters or a planter who had a significant amount of money for capital. In Barbados the average holding was 150 acres after the change to sugar.

If it was below this amount, then the estate tended not to e profitable. About half of the area was under sugar; a sixth would be for the cattle, another sixth for growing crops such as vegetables and fruits and the remainder for woodland which would be used for timber and firewood. When the sugar revolution was undergo it caused the price of the land to become exceeding high and in some parts of Barbados by as much as thirty times. For instance in 1630 the average price of an acre was three pound (E). By 1648 when the sugar revolution was almost complete in Barbados, an acre was sold for over thirty pounds (EYE). By Imaginable

All cells need to have a constant energy supply. The two processes by which this energy is attained from photosynthetic materials to form ATP are cellular respiration and fermentation. (Hyde,2012). Fermentation is a way of harvesting chemical energy that does not require oxygen. (Reece et al. 2012). When the body is deprived of oxygen it will then begin to meet its energy needs through the slow process of fermentation. In our lab we investigated alcoholic fermentation by using yeast, which can flourish in an low energy environment in anaerobic conditions.

In this lab our goal was to discover the rate at which yeast will ferment different sized molecules of carbohydrates. In order to perform our experiment we made use of water, glucose, sucrose, and starch. It was hypothesized that glucose, sucrose, then starch would all be used to produce energy during fermentation. Being that glucose is a simple sugar, or monosaccharide, we predicted that glucose would be fermented most quickly. This hypothesis was made based on the idea that glucose is the cell’s main source of energy in aerobic cellular respiration. The first step of cellular respiration is glycolysis which breaks down glucose for energy.

We predicted that Sucrose would ferment second to glucose since it is a larger molecule composed of glucose and fructose. Finally, we predicted that starch would ferment extremely slow behind all of the other carbohydrates. METHODS AND MATERIALS: On October 31, 2012 in the lab of Greenfield Community College my lab partners, Madeline Hawes, Timothy Walsh and I conducted the following experiment in order to test the effectiveness of yeasts’ ability to ferment different carbohydrates. We first filled 6 small flasks with 75 ml of water and 5 drops of phenol red to each flask.

Four of these were labeled with the solution that would feed into them and the other two with “control” and the last with “increased CO2. ” The color of phenol red is orangish-pink when there is a neutral pH present. As carbon dioxide is released into this solution from the release of the gas from the yeast filled flasks, the solution turns a light yellow indicating a weak acid. We measured out four weigh boats of 2 grams each of starch and then added 2 grams to each of 4 labeled flasks of 50 ml water, 50 ml Glucose solution, 50 ml Sucrose solution, and 50 ml Starch solution respectively.

All of these had been stored in incubators to maintain an optimal temperature of 35 degrees celsius. We put these flasks into our sink which we made into a water bath. We then drained and added hot plate warmed water from a 1000 ml beaker we kept heated in order to maintain the optimal temperature of 35 degrees celsius around the flasks. We swirled the large flasks to mix the solutions and yeast as they sat in the water bath. The flasks containing the yeasts solutions were then stoppered with glass straws and tubings and their extending tubes placed into the matching labeled smaller flasks adjacent to the sink.

I blew through a straw into the flask labeled “increased CO2. ” The phenol red detected the presence of CO2 turning the solution yellow. The “control” flask was left as a comparison for the remaining yeast filled tubes feeding into the other flasks of phenol red and water. RESULTS: We recorded our first observations at 10 minutes. Just as we hypothesized, the yeast and water experienced no change. In the glucose solution flask, the glucose molecules were being quickly broken down and forming a frothy head, sending a bubble of CO2 through the tube every 2 seconds while turning the phenol red to a light orange.

The sucrose solution was bubbling every three seconds and also had turned light orange. At 10 minutes there was no reaction in the Starch solution. The latter data remained consistent with our hypotheses. The glucose solution at 20 minutes was very frothy and bubbly and had turned the phenol red a very light yellow with a consistent bubble through the tube every second indicating a strong presence of CO2. The sucrose, too, had turned light yellow and had continuous bubbles every 2 seconds. The starch had a rare bubble with no noticeable change in the phenol red solution.

At the final check in of 40 minutes both the glucose and sucrose had fermented most of the yeast and slowed down on bubbling. The glucose still had the most bubbles occuring. The starch was a lighter pink with little change in the levels of froth in the yeast solution. The water solution still remained completely unchanged. DISCUSSION: Our hypotheses were supported through illustrating that all forms of sugar do provide energy and that glucose, being the smallest molecule, was the most efficient. The control tube contained no sugar and therefore produced no energy. A source of sugar is necessary for glycolysis and fermentation to occur.

The strongest presence of carbon dioxide was in glucose, indicated by the bubbles which are a by-product of ethanol fermentation. The rate of fermentation in sucrose was second to glucose and Starch was the least effective at providing a sugar to create energy. The large polysaccharide was difficult for yeast to break down to create the necessary energy that would produce carbon dioxide. Glucose is the most efficient sugar as it is a small monosaccharide which is already the source of energy for the Glycolysis cycle. The largest possible source of error in our experiment is the time in which each solution began its fermentation process.

We added the yeast into each flask containing the sugar solutions at staggered times. If this experiment were to be repeated it would be more precise to have four people pour in the yeast and swirl at the exact same time and then stopper the solutions. The only minor inconsistency would be the amount of yeast that was spilled or left in the weigh boats. This could create a discrepancy in the final results. Through this lab I understoodd that in times of oxygen deprivation the body can still function through the process of fermentation.

The yield of 2 ATP molecules is enough to keep muscles contracting for a short period of time when oxygen is scarce. Through the fermentation process NAD+ is regenerated as pyruvate is broken down to CO2 and ethanol. This allows the anaerobic production of 2 ATP molecules. (Reece et al. 2012). In essence, keeping cells alive that may otherwise die without the energy to provide for muscle contractions of the heart.

LITERATURE CITED: Reece, Taylor, Simon, Dickey, and Campbell. , Biology: concepts & connections. Pearson Benjamin Cummings, San Francisco, CA. Pgs. 100-101 Hyde, A. October 31, 2012

Yeasts’ capability of undergoing ethanol fermentation, its ability to ferment other sugars and artificial sweeteners, and how lactase influences yeasts ability to use lactose as a food source Kristina Naydenova Father Michael Goetz Purpose Part A: To investigate whether yeast has the ability to ferment glucose to produce carbon dioxide gas and ethanol. Part B: To investigate whether yeast has the ability to ferment other sugars and artificial sweeteners and how lactase influences their ability to use lactose as a food source. Question

Part A: Does yeast have the ability of undergoing ethanol fermentation? Part B: Does yeast have the ability to ferment other sugars and artificial sweeteners? Does lactase influence the ability of yeast to use lactose as a food source? Hypothesis Part A: If yeast produces carbon dioxide gas (the solution will turn cloudy due to carbon dioxide presence) and ethanol after fermenting glucose then it has the ability to undergo ethanol fermentation because ethanol fermentations reactants consist of glucose and the products consist of carbon dioxide gas and ethanol.

Part B: If yeast has the ability to ferment other sugars and artificial sweeteners then the products of the solution will consist of carbon dioxide gas and ethanol because the products of ethanol fermentation are carbon dioxide gas and ethanol. If lactase influences the ability of yeast to use lactose as a food source then the yeast will be able to use lactose to produce carbon dioxide gas and ethanol because the yeast will be capable of breaking down lactose into glucose and galactose. Materials * Safety goggles| * Lab apron| 4 flasks (100 mL) and 1 stopper| * Wax pencil (for making test tubes)| * Ruler| * 6 large beakers (400 mL)| * Thermometer| * Stopwatch| * 50 mL glucose suspension (10%)| * 50 mL yeast suspension (I package per 100 mL of water)| * Cotton batting| * Limewater| * Warm water (35 °C)| * 10 mL of each of the following solutions: glucose, sucrose, lactose, and artificial sweetener (10%)| * 10 mL of a suspension of lactose (10%) with a pinch of lactase| * 10 mL of distilled water| * Graduated cylinder| * 6 test tubes (15 mL) with 1 hole rubber stoppers|

Variables Procedure 1. The safety goggles and lab apron were put on. 2. Three flasks were labeled as “yeast and glucose,” “yeast,” and “glucose. ” 3. 10 mL of glucose solution and 5 mL of yeast suspension were added to the “yeast and glucose” flask. 4. 10 mL of distilled water with 5 mL of yeast suspension to the “yeast and glucose” flask as a control. 5. 5 mL of distilled water with 10 mL of glucose solution were added to the “glucose” flask as a second control. 6. Cotton batting was placed in the mouth of the flasks to reduce air turbulence. . The cotton batting was removed carefully after 24 hours and the contents of each flask were smelled. A slight alcohol odour was detected. 8. Each flask was tested for the presence of carbon dioxide.

The invisible gas mixture was slowly poured into a flask that contains 25 mL of limewater. The limewater flask was stoppered and the contents were swirled to mix the limewater with the gas. Observations were recorded. The flask was rinsed. 25 mL of fresh limewater was added before testing the next gas sample. . A ruler was used to place graduation marks at 0. 5 cm intervals along the sides of the test tubes. 10. Six beakers of warm water (35 °C) were prepared. The beakers were two-thirds full of warm water. 11. The six test tubes were labeled as “glucose,” “sucrose,” “lactose free milk,” “artificial sweetener (Splenda),”lactose free milk and lactase,” and “distilled water”. 12. 10 mL of the appropriate solutions to each test tube were added. 13. 5 mL of yeast suspension to each test tube were added.

The test tubes were filled. 14. The test tubes were scaled with one-hole stoppers after the mixtures are placed in the test tubes. 15. One test tube was held. The holes in the stopper were covered, and the test tube was inverted and placed into a beaker of warm water. The process was repeated for all six solutions, using a different beaker for each solution. 16. The amount of gas produced after 1, 5 and 10 minutes was recorded using the graduation marks on the test tubes. Observations Table 1.

Before and after observations of yeast and glucose, yeast and glucose Solution| Before| After| Yeast and glucose| | | Yeast| | | Glucose | | | Table 2. The amount of gas produced by glucose, sucrose, lactose free milk, artificial sweetener, lactose free milk and lactase, and distilled water after 1, 5 and 10 minutes Solution| Time (1 minute)| Amount of gas| Time (5 minutes)| Amount of Gas| Time (10 minutes)| Amount of Gas| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |

Title of the Experiment: Enterobacteriaceae Identification: The Enterotube II System Learning Objectives: After completing this exercise we were able to inoculate an unknown bacterium that belongs to the Enterobacteriaceae by using technology effectively with a Enterotube II. An Enterotube II is a miniaturized multi-test system for rapid identification of enterbacteriaceae. We then evaluated the test results and generated a five-digit code for the unknown bacterium.

Thinking creatively and critically we had to fgure out the code by looking at the Enterotube and comparing he reactants to the original to see what the result was. We then had to use the five- digit code to correctly identify the unknown bacterium from the interpretation guide. Procedure: Step 1 : Remove organisms from a well-isolated colony. Avoid touching the agar with the wire. Step 2: Inoculate each compartment by first twisting the wire and then withdrawing it all the way out through the 12 compartments using a turning movement.

Step 3: Reinsert the wire (without sterilizing), using a turning motion through all the 12 compartments until the notch on the wire is aligned with the opening of the tube. Step 4: Break the wire at the notch by bending. The portion of the wire remaining in the tube maintains anaerobic conditions essential for true fermentation. Step 5: Punch holes with broken-off part wire through the thin plastic covering over depressions on sides of the last eight compartments. Replace caps and incubate at 35 degrees C for 18-24 hours.

Step 6: After encircling the numbers of the positive tests on the laboratory report, total up the numbers of each bracketed series to determine the five-digit code number. Refer to the Enterotube II Interpretation Guide for identification of the unknown by using the code number Results: Before inoculation of the Enterotube II showing the original colors of each test which was also used to compare with the inoculated enterotube. After the inoculation of the Enterotube, many of the colors have changed which means they have reacted with the antibiotic.

The reactants then helped me find out what the unkown bacterium is. Each color changed or reacted gives a certain digit Summary & Conclusions: Enterotube II identifies Enterobacteriaceae. The Enterotube II is a multiple test system designed to identify enteric bacteria based on Glucose, Adonitol, Lactose, Arabinose, Sorbitol, Dulcitol fermentation, lysine and Decarboxylation, Sulfur reduction, Indole, Acetoin production of glucose fermentation, Phenylalanine deamination, Urea hydrolysis, and Citrate utilization.

Adonitol Bacterial fermentation of adonitol, which results in the formation of acidic end products, is indicated by a change in color of the indicator present in the medium from red (alkaline) to yellow (acidic). Any sign of yellow should be inter preted as a positive reaction, orange should be considered negative. Lactose Bacterial fermentation of lactose, which results in the formation of acidic end roducts, is indicated by a change in color of the Indicator present in the medium from red (alkaline) to yellow (acidic).

Any sign of yellow should be interpreted as a positive reaction; orange should be considered negative. Arabinose Bacterial fermentation of arabinose, which results in the formation of acidic end products, is indicated by a change in color from red (alkaline) to yellow (acidic). Any sign of yellow should be interpreted as a positive reaction; orange should be considered negative. Sorbitol Bacterial fermentation of sorbitol, which results in the formation of acidic nd products, is indicated by a change in color from red (alkaline) to yellow (acidic).

Any sign of yellow should be interpreted as a positive reaction; orange should be considered negative. Voges-Proskauer Acetylmethylcarbinol (acetoin) is an inter mediate in the production of butylene glycol from glucose fer mentation. The presence of acetoin is indicated by the develop ment of a red color within 20 minutes. Most positive reactions are evident within 10 minutes. Phenylalanine Deaminase This test detects the formation of pyruvic acid from the deamination of phenylalanine. The pyruvic acid formed reacts with a ferric salt in the medium to roduce a characteristic black to smoky gray color.

Urea The production of urease by some bacteria hydrolyzes urea in this medium to produce ammonia, which causes a shift in pH from yellow (acidic) to reddish-purple (alkaline). This test is strongly positive for Proteus in 6 hours and weakly positive for Klebsiella and some Enterobaeter species in 24 hours. Citrate Organisms that are able to utilize the citrate in this medium as their sole source of carbon produce alkaline metabo lites that change the color of the indicator from green (acidic) to deep blue (alkaline). Any degree of blue should be considered positive.

After looking at the results of the Enterotube I came to conclusion that my unkown bacteria gave me the five-digit code 34363 which translated to Klebsiella pneumonia bacterium that can form a capsule. It is found in the normal flora of GI tracts in humans. K. pneumoniae can become pathogenic in patients whose immune systems are compromised. K. pneumoniae can cause nosocomial urinary tract infections and pneumonia. In immunocompromised patients, death is possible. For a personto get the K. pneumoniae bacteria, they have to have direct contact with another person. K. pneumoniae is not able to be contacted through the air.

Healthcare workers can help to decrease the spread of K. pneumoniae by washing their hands before and after taking care of a patient. It was established that the Enterotube system provides a simple, reliable, and rapid method for the probable identification of Enterobacteriaceae. The major advantage of the Enterotube is that all tests are done simultaneously by inoculation from a single isolated colony. It is also easier to inoculate, single inoculation, self- contained, numerous tests, little media preparation, rapid results, reliability, uniformity, simple interpretation.

In view of the commercial importance of black tea and the intricacy of the mechanisms of its manufacture, this product has received by far the most attention and the purpose of the present article is to outline some findings in this field. The black tea process

1. The freshly plucked tea flush is allowed to wither in air for some 18-20 hours, or for shorter periods when heated air is circulated, when it loses water and acquires a kid-glove feel. Important chemical changes have already begun to take place
2. For example, amino acids are formed as precursors of compounds ultimately leading to the production of flavor and non-enzymes browning, the formation of kite compounds as flavor precursors and the 2 formation of caffeine. The leaf also becomes capable of acquiring a twist, rather than breaking up, when it is subsequently rolled. Fermentation is initiated by rolling when the enzyme, normally located in the chloroplast, and the phenol substrate, found in the cell vacuoles, are mixed in the presence of oxygen, without extensive damage to the outer cell wall.A three hour fermentation results in less than 10% of unchanged substrate remaining.
3. Fermentation is arrested by firing in a stream of hot air which also dries the product to some 3% moisture content. The final stage is grading. Enzymes oxidation Phenols or polysaccharides are enzymes which mediate in the oxidation of o-depletion to o-quinine’s in the presence of oxygen but most of these enzymes are also capable of oxidation monopoles to o- quinine’s. The tea enzyme is a polysaccharides but, unlike the ordinary for the so called fermentation are flavor components of the tea leaf. These are based on the flan structure. Polyphonic components comprise some 25-35% of the tea flush on a dry weight basis, of which some 20% may be found as flavor
4. Specific flavor structures are shown in figure 2. They may clearly be divided into two groups ? the catechist and the collocations according to whether there are two or three hydroxyl (OH) groups in the right hand phenol ring. In fact, each group of compounds may be further distinguished according to the arrangements of groups around carbon atoms 2 and 3, resulting in four possible isomers.

For example, the isomers of the catechisms are:

1. (-) catechist,
2. (+) catechist,
3. (-) peachiness
4. (+) peachiness.

In addition, these compounds exist as esters with Gaelic acid, figure 3. The most abundant are the collocations and specifically (-) epistemologically and its gallant ester (ca. 10% dry weight). In order of abundance, this is followed by (-) peachiness and its gallant (ca. 5 by weight)

It is reasonable to assume that the first stage of oxidation involves conversion of Nutrition and Food Science these substrates to o-quinine’s and is followed by condensation of these quinine’s to dimmers and polymers. Flavor derived products in black tea The oxidation of flavors by way of quinine leads to the formation of dimmers by meaner of bonds between adjacent molecules, such that the 2′ position on one molecule, figure 2, links to either the 6 or 8 position on another in the case of catechist (ahead to tail’ dimmers), and in the case of collocations the 2′ position on one molecule becomes linked to the 2′ position on another (tail to tail’ dimmers). These tail to tail dimmers have been identified in black tea and are found to be derived from (-) epistemologically and its gallant as expected

5. During fermentation carbon dioxide is evolved and this is believed to arise from an unusual but most important reaction leading to the formation of a seven member ring. Carbon rings of this size are infrequently found in organic chemistry but the essentials of this reaction are illustrated by the oxidation of paroxysmal to form purloining, figure 4. Gaelic acid, found extensively in fermented tea, can undergo a similar reaction to form purpurogallincarboxylic acid. The thyroxin grouping of the collocations can react in a similar manner to paroxysmal and it is therefore, not surprising that compounds such as paleontologist, figure 5, are found to be present in black tea

6 . It is also found that the catechist can take the place of one molecule of reactant in the purloining reaction. Thus, catechist can react with Gaelic acid to form diphtheria acids, figure 6, but, more importantly, one molecule of catechist is capable of reacting with one molecule of collocating, again in a purloining type reaction 2 . The product is known as deflating and the structure is shown in figure

Deflating and its gallant esters are very important orange-red coloring matters in black tea constituting some 2% by weight on a dry basis. However, by weight, the most important group of coloring matters in black tea is that known as therapeutics constituting more than 10% . Their structure is still unknown but they may also Evaluation of tea Tea is evaluated under five headings: strength, color, briskness, aroma and quality.

Strength is a measure of the total concentration of deflations and therapeutics and, since they are responsible mainly for the color of tea, with small contributions from paleontologists and products of November 1979 3 TEA continued non-enzymes browning, color and strength are related. However, the assessment of color is more a measure of the brightness of the color rather than total color and so is a measure of the balance between the deflations and therapeutics, the former contributing sensory brightness and the latter the depth

The extent of popularization of tea polyphony’s depends on such factors as time and temperature, more extensive popularization giving rise to reduction of solubility. The polymers combine readily with caffeine and the result on cooling is known as creaming, the compounds so formed tending to separate out. This is particularly undesirable in teas intended for making iced tea. Creaming can be assessed through the cream index which is determined by deliberate coagulation with acid.

The astringency of tea is largely dependent on the amount of polyphonic compounds present, the degree of oxidation of the tea flavors and particularly by the amount of Gaelic acid groups present on the flavors and their oxidation products. Caffeine is reported to improve the briskness of tea and milk or lemon Juice may modify the taste of the polyphony’s 2 . The overall quality of a tea infusion may also be related to he proportions of deflating and therapeutics present and also to the sum of their concentrations.

The aroma of tea is not related to tea polyphony’s but is determined by the volatile components. Some three hundred compounds have been identified in black tea and recent discoveries are listed in the latest review 2 . They comprise leaderless, stones, esters, pyridine’s, paralyzes, thistles, squishiness, aromatic amines, amides and other compounds. The formation of carbonyl compounds is a result of Stretcher degradation reactions between amino acids and oxidized flavors according to: usability stresses the importance of the formation of amino acids during the withering stages of tea manufacture.

Tea leaves, being photosynthetic organs, also contain a significant amount of cartooned and important black tea aroma components are probably produced as a result of the oxidative degradation of carotids. The oxidation of unsaturated fats may also contribute to flavor. Conclusion The most important stage of black tea manufacture involves enzymes oxidation of flavor substrates. Demerit flavors and particularly deflating are important contributors to tea quality together with the higher polymers known as heartburning. It is worth noting, however, that condensation does not stop when the enzyme is inactivated during firing.