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
Data from Table 1. confirms the theory that as the concentration of glucose increases so will the absorbance of the solution when examined with the glucose oxidase/horseradish peroxidase assay. Glucose within the context of this assay is determined by the amount of ferricyanide, determined by absornace, which is produced in a one to one ratio.1 Furthermore when examining the glucose standards, a linear calibration curve was able to be produced (shown as Figure 1). Noted the R2 value of the y = 1.808x - 0.0125 trend line is 0.9958, which is statistically considered linear. From this calibration curve the absorbance values of unknowns samples can be compared, and the correlated glucose concentration can then be approximated.
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
High-fructose corn syrup (HFCS) is an artificial sweetener commonly used in the United States. As its name implies, this sweetener is derived from agricultural corn. All high fructose corn syrups are corn syrups whose fructose content has been increased via enzymatic processes and then mixed with pure corn syrup. There are several different formulations of high-fructose corn syrup. The product sold in the United States (HFCS #2) has the following composition: moisture, 29%; dry substance, 71% D.S.; dextrose, 50% D.S.; ash, 0.03 D.S.; and nitrogen, 0.002% D.S. The amounts of dextrose, fructose, and other saccharides may vary slightly in HFCS #3, but the analysis is fairly consistent. HFCS #1 hasn’t been commercially sold specifically for consumer consumption in the U.S. for many years. Instead, it is used by food producers in their products.
These labels indicated the lactose solution that was be placed into the mini-microfuge tubes. The varying lactose ph solutions were obtained. The four miniature pipets were then used, (one per solution,) to add 1mL of the solution to the corresponding mini-microfuge tubes. When this step is completed there were two mini-microfuge tubes that matched the paper towel. Then, once all of the solutions contained their respective lactose solutions, 0.5mL of the lactase enzyme suspension was added to the first mini-microfuge tube labeled LPH4 on the paper towel, and 4 on the microfuge tube. As soon as the lactase enzyme suspension was added to the mini-microfuge tube, the timer was started in stopwatch mode (increasing.) When the timer reached 7 minutes and 30 seconds, the glucose test strip was dipped into the created solution in the mini-microfuge tube for 2 seconds (keep timer going, as the timer is also needed for the glucose strip. Once the two seconds had elapsed, the test strip was immediately removed, and the excess solution was wiped gently on the side of the mini-microfuge tube. The timer was continued for 30 addition seconds. Once the timer reached 7:32 (the extra two seconds accounting for the glucose dip), the test strip was then compared the glucose test strip color chart that is found on the side of the glucose test strip
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
Finally, the last part of the experiment examined the enzyme activity at different pH levels. Four sets of 11 tubes were set up in this part. The procedure for this part is the same as before, but 4 other buffers were substituted for the standard pH 7.3 phosphate buffer. Set A used the 5.5 pH buffer while set B used the 6.5 pH buffer. The buffer of pH 8.5 was used for set B and for set D the pH was 9. The absorbance readings for 4 sets were taken and recorded in table 13. Using the linear equation that the best-fit line gave for each set, the Km and the Vmax of each set were determined. Then, table 15 was made by dividing the Vmax by the Km. of the four pHs. The Vmax and Km of the control set were also used to make
PH can affect the way fermentation occurs due to the chemical differences between acid and alkaline elements, particularly within a solution. In this experiment an enzyme-based reaction was examined that in order to observe this pH trend. The aim of the experiment was to determine how pH affects the yeast fermentation rate by performing the experiment numerous times with a different pH (pH's 3, 5, 7, 9, 11) in different glucose solutions. The hypothesis was ‘If the pH is lower than the neutral point, then the fermentation reaction will occur faster?.’ The experiment conducted was to measure the amount of carbon dioxide (C02) produced by the yeast during fermentation whilst modifying the pH of the glucose solution. To test this every 5 minutes
This experiment involves performing various techniques, including heating under reflux, separation, drying, distillation, gas chromatography (GC), infrared spectroscopy (IR spectroscopy), and nuclear magnetic resonance (1H NMR). Heating under reflux is important to overcome any activation barrier of energy that may be present in order to complete the reaction.
Tappy, L., Lê, K. A., Tran, C., & Paquot, N. (2010). Fructose and metabolic diseases: New findings,
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
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.
Further, fermented food itself is interpreted by Campbell-Platt (1987) “as those foods which have been subjected to the action of micro-organisms or enzymes so that desirable biochemical changes cause significant modification to the food.” (As cited in Sahlin, 1999, p.5)
The sample was subjected to steam distillation as illustrated in Figure 1. A total of 50ml of distillate was collected while recording the temperature for every 5.0 ml of distillate. The distillate was transferred into a 250ml Erlenmeyer flask and 3.0 g of NaCl was added. The flask was cooled and the content was transferred into a 250-ml separatory funnel. Then 25.0ml of hexane was added and the mixture was shaken for 5 minutes with occasional venting. The aqueous layer was discarded and the organic layer was left inside. About 25.0ml of 10% NaOH was then added and the mixture was shaken as before. The aqueous layer was collected and then cooled in an ice bath. It was then acidified with enough 6.00 M HCl while the pH is being monitored with red litmus paper. Another 25.0 ml of hexane was added and the mixture was shaken as before. The hexane extract was saved and a small amount of anhydrous sodium sulfate was added. The mixture was then swirled for a couple of minutes then filtered. A small amount of the final extracted was tested separately with 1% FeCl3 and Bayer’s reagent.
Add 15mL of 6N sulfuric acid to a 125mL Erlenmeyer flask containing 105mL of deionized water (preparing approximately 0.75N sulfuric acid). Obtain a sample of the unknown. Weight the vial and contents accurately on an analytical balance. Handle the vial with a small strip of paper to reduce the risk of error (due to added weight). Pour about half of the sample into a clean dry 200mL Erlenmeyer flask and weight again. Use the remaining half of the sample to get a second weight of around 0.6g-0.7g. Make sure the vial is capped on every weight taken.
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