Experimental Setup: Fermentation Broth Analysis Fermentation broth was diluted by a 1:5 dilution to 40 grams. After dilution the sample was centrifuged at 4000 rpms for 5 min. The supernatant was filter sterilized using .22micron filters. After filtering, the sample was analyzed using HPLC with a BioRad Aminex HPX-87H Column. Using this method, it was able to be determined the amount of Maltose, Glucose, Fructose, and Maltotriose in each sample. Enzyme Testing in Broth Enzyme testing was performed in 20mL scintillation vials using diluted broth made for analysis and a range of enzyme concentrations of 1ml of enzyme/500 ml of broth, to 1ml of enzyme/500L of broth. Using 1.5 ml of diluted fermentation broth and 1.5 mL of enzyme solution, the …show more content…
This was done by a Trichloroacetic acid (TCA) precipitation. Using 250 grams of solid TCA and adding 175 ml of water gave us a solution for precipitation. However, this solution had to be diluted 1:40 due to detector flooding on the RI detector of the HPLC. 1mL of Diluted was added to each reaction sample, and incubated at 3°C for 10 min. After incubation, the sample was centrifuged at 9.6K rpm for 10 min. Sample was then filtered with a .22μ filter before being loaded into HPLC for analysis. HPLC Method The HPLC method used was 20 μL of sample at 25°C through an RI detector for 15 minutes. The mobile phase was .0001M Sulfuric acid at .6ml/min with a column temperature of 60°C. Findings and Discussion: Preliminary Findings When samples first arrived, dilutions were made and tested on the HPLC. Table two has the resulting concentrations of glucose, maltose, maltotriose and fructose. Using this data, the $0.45 per kg price of Clearsweet, and the moisture percent of 29%, an average potential savings was created for each batch. This potential cost savings is the greatest for DHA-T and DHA-T HP at an average of $4,500 per run due to un-fermentable sugars. Natural beta carotene has a drastically greater amount of fructose from the sample …show more content…
Table 4. Enzyme Experiment at 20°C for 24 hrs on P5277 DHA-T HP Table 5. Enzyme Expirement at 20°C for 24 hrs on M146 DHS Fructose Conversation in UHTS Experimental Setup: DE 95 Clearsweet Corn syrup was ran through the Winchester Pilot Plant’s ultra high temperature sterilizer (UHTS) At tempertures ranging from 130-140°C and volumetric flows ranging from 7L/min to 5L/min Results of Exeriment: Table 6. Fructose Generation experimental results. Since the results from Table 6 show a small difference between fructose amounts before dilution, the reaction results were inconclusive due to our HPLC method and accuracy. If an internal Std. was added, the results may be better. However, we can conclude that the fructose was consumed in the
If this experiment were designed to determine the amount of Fructose in a solution, describe what, if anything, would need to change in the reaction? Explain why there would or would not need to be changes. (5
The isolate possesses some enzymes required for hydrolytic reactions. Hydrolytic enzymes found to be secreted from the bacterium, are -amylase, casein, and PYRase. In the starch hydrolysis and casein tests, there was a zone of clearing around the bacterium, which was indicative of the secreted enzymes necessary to break down starch and casein. In the PYR test, the presence of PYRase was detected by a color change to red on the PYR disc after the addition of the PYR reagent (p-dimethylaminocinnamaldehyde). Hydrolytic enzymes for which the EI tested negative were urease, gelatinase, and DNAse. In the Urea Hydrolysis test, it was observed that the urea broth did not have a color change, indicating that there was no urease secreted to break down urea in the broth. Similarly, there was no gelatinase present to break down gelatin in the Gelatin Hydrolysis test, so the nutrient gelatin remained solid. It was concluded that the EI does not possess DNase because there was no clearing zone around the bacteria, indicating that DNA had not been
Forristal, Linda. “The Murky World of High-Fructose Corn Syrup.” Westonaprice.org. Ed. Linda Forristal. 2003. 24 July 2008. .
While the tube for specimen Cb turned a tannish white in the lower half of the tube while the top stayed the lavender inoculated tube color. Do to this evidence I determined that both specimens Ca and Cb cannot use the process Casein hydrolysis or Casein coagulation due to lack of soft or firm curds in both tubes. Since there was no casein curds formed, I concluded that specimens Ca and Cb also cannot perform the process of proteolysis. My conclusion is supported by the fact that there was no clearing of the medium. I have also determine that neither of my organisms can make the enzymes rennin, proteolytic or even proteases. I know my specimens cannot produce proteases due to the fact that there was no blue coloring in the tubes which means that the byproduct Ammonia was not produced to increase the pH. Since neither of my specimens can make these enzymes, I concluded that my specimens cannot break down lactose or casein. Although I did learn that specimen Cb can reduce litmus due to the evidence that the lower part of the tube turned a tannish white color with a purple ring at the top. This color change from a purple to a white means that the litmus was reduced turning it clear and leaving the white of the milk to show. Finally I know that specimen Ca cannot reduce litmus due to the fact that the tube had no change in
High fructose corn syrup was first created in the 1970s by the Japanese as a form of sweetener. Combining 45% glucose and 55% fructose it was the sweetest substance yet and its cheap production, longer shelf-life, and versatility helped it over the next three decades emerge as the dominant sweetener on the market. However, despite its success, it has most recently been noted that effects of the substance are extremely detrimental to consumers, and its increased use directly correlates to the rise in obesity and diabetes among Americans.
Abstract: High fructose corn syrup (HFCS), like many other unhealthy constituents that are used in foods, is cheap and retains the taste of the natural products it emulates, possibly even surpassing them in many areas. However, experiments have shown that fructose is not an ideal sugar for human consumption, not to mention the fact that the use of GM ingredients can be dangerous. In order to prevent the continued consumption of this noxious sugar, food producers should use healthy alternative sweeteners to prevent the further dependence on HFCS in our foods and drinks.
We hypothesized glucose would produce the most carbon dioxide, followed by Corn syrup, fructose, and lastly sucrose. Glucose would yield the highest amount carbon dioxide because it is normally the sugar catabolized during fermentation. Corn syrup was next because it is a mixture of both glucose and fructose. Fructose is a monosaccharide and therefore would have less catabolism involved than sucrose, a disaccharide. This means that the fructose may go through fermentation faster within the 20 minute period and produce more carbon dioxide. We then predicted the test tube containing glucose would have the highest amount of carbon dioxide at the end of the 20 minute period, followed by corn syrup, then fructose, then sucrose. Our initial prediction of glucose yielding to the highest level of carbon dioxide produced was supported by the experiment. However, corn syrup yielded a lower average amount of carbon dioxide than fructose and sucrose (figure 1). This deviation from our hypothesis and prediction may be due to inadequate amounts of agitation of the test tube during the 2 trials involving the corn syrup.
As a group the slope of the glucose line was recorded as 59.001 ppm/min. The class average for glucose was conducted from numbers that were varied in size. The lowest number being 3.0136 ppm/min and the highest being 1026.2 ppm/min. The class average was calculated as 471.201 ppm/min for the respiration rate of yeast when metabolizing glucose. Using class averages from multiple periods the result of 679.48 ppm /min was collected as the overall average. Moreover, the collection of data for sucrose was also compressed into averages. The slope of the line for sucrose was recorded as 280.3 ppm/min for a group collection. The data of sucrose included numbers starting at -873.51 ppm/min and ended with 1522.8 ppm/min being the highest. The class average recorded for the respiration rate of yeast when metabolizing sucrose was 649.246 ppm/min. The overall average of sucrose was 575.686 ppm/min. Also, the data for starch and lactose were also calculated into class averages and overall averages. As a group the slope of the lactose line was recorded 155.69 ppm/min, and the slope of the starch line was recorded as 367.34 ppm/min. The class average calculated for lactose was 214.183 ppm/min while the average for starch was 389.439 (units). Lastly, the overall averages were collected for lactose and starch. The overall average of lactose
HFCS is a popular sweetener used in processed foods. It is composed of approximately 50% fructose and 50% glucose. It is made from corn starch with the use of enzymes to convert glucose to fructose. It has many advantages over cheap sugar, including, but not limited to, lower price, longer shelf life, low freezing point, and enhanced taste and texture. Corn refinement was first discovered circa 1860, and was soon followed by the development of corn syrup. Important advantages took place in the 1920’s with the use of enzymes, but it was not until the mid-1900’s when the crucial glucose isomerase enzyme was discovered. Industrial production of HFCS began in the 1970’s and today the industry is huge.
Molisch test is one of the useful qualitative test for presence of carbohydrates in solution. The three glucose solutions all have a violet colored ring formed at the junction between the two layers. This showed that carbohydrates are present in these sugar solution. This test involved the addition of concentrated sulphuric acid which causes dehydration of all carbohydrates to give ‘furfural’ compound, where pentoses are dehydrated to furfural, and hexoses are dehydrated to 5-hydroxymethylfurfural (Molish's reagent, 2009). These compounds will later react with – naphtol which is Molisch reagent to give a purple colored complex. The test has to be carried out slowly as the violet colour formed at the surface of contact of concentrated sulfuric
Materials and Methods: An ion exchange chromatography column was obtained and set up for purification with the addition of 0.5 ml ion exchange matrix. 1 ml
Steinkraus, K. H. (2002). Fermentations in world food processing. Comprehensive Reviews in Food Science and Food Safety, 1(1), 23-32.
By taking a Carbon Dioxide, rich substance and mixing it with a yeast, solution fermentation will occur, and then it could be determined if it is a good energy-producer. In this study glacatose, sucrose, glycine, glucose, and water were used to indicate how fast fermentation occurred. The overall result shows that monosaccharides in particular galactose and glucose were the best energy source for a cell.
Careful note: Pour all of the liquids used in this experiment in a fume hood.
Glucose is a monosaccharide with formula C6H12O6. It occurs in Free in the ripen grapes in bones and also in many sweet fruits. It is also present in human blood to the extent of about 0.1%. Sucrose is one of the most useful disaccharides in our daily life. It is widely distributed in nature in juices, seeds and also in flowers of many plants. The main source of sucrose is sugar cane juice which contain 15-20 % sucrose and sugar beet which has about 10-17 % sucrose. The molecular formula of sucrose is C12H22O11. It is produced by a mixture of glucose and fructose. It is non-reducing in nature whereas glucose is reducing. Soft drinks are a bit acidic in nature and their acidity can be measured by finding their pH value. The pH values also depend upon the acidic contents such as citric acid and phosphoric acid.These drinks are readily consumed on daily bases especially when undergoing tedious activities. Also, with the relatively affordable prices, they are highly consumed during leisure and relaxation outings and serve the general public in