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Fermentation Broth Analysis: Analysis Of Chemistry In Chemistry

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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
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