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Title Theory/Introduction Methodology/Procedure Process Flow Diagram (PFD) Workbook/Reports Results TOTAL MARKS

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------------------------------(DR. JAGANNATHAN KRISHNAN) Date:

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Fermentation is an energy yielding process that converts carbohydrate into acids or alcohol by using bacteria and yeast. The substrates that are commonly used are glucose (sugar) and amino acids. Typical products of fermentation are organic acids (lactic acid, acetic acid), alcohol (ethanol, methanol, buthanol), ketones (acetone) and gases (hydrogen and carbon dioxide). The industrial production by fermentation can be beverages (beer and wine), dairy product (milk and cheese), breads and bioproducts (biofuel and bioethanol). There is more research done on the production of bioproducts nowadays as a renewable energy since crude oil is depleted. The amount of the bioproducts depends on the sugar used. Researchers are also improving the fermentation technologies and Gene-modified yeast is even used for a higher efficiency. Bacteria such as Streptococcus, Lactobacillus, and Bacillusis used to produce lactic acid in certain food. Yeast contains enzyme (glycolytic enzyme) that act as catalyse to breakdown the glucose into ethanol and carbon dioxide. In the absence of oxygen (anaerobic fermentation), the cell primary mechanism to produce adenosine triphosphate (ATP) is glycolysis. Glycolysis transfer electrons to NAD+, forming NADH. NADH must then be oxidized to regenerate the NAD+ since NAD+ supply is limited in any cells. Glucose molecule is split into 2 pyruvate molecules. The net gain of 2 ATP and 2 NADH are produced, as well as water molecules via substrate-level-phosphorylation. C6H12O6 + 2 ADP + 2 Pi + 2 NAD+ 2 CH3COCOO + 2 ATP + 2 NADH + 2 H2O + 2H+ (glucose) (inorganic phosphate) (pyruvate)

Figure 1: fermentation process The five main purposes of food fermentation are: a. To enrich flavors, aromas and textures in food substrate. b. To preserve substantial amount of food through lactic acid, alcohol, acetic acid and alkaline fermentation. c. For biological enrichment with protein, essential amino acids, essential fatty acids and vitamins while eliminating the antinutrients. d. To decrease cooking time and fuel requirement.



1. Select Task, pure components and register,edit/view properties to register all the components into the process which are media, oxygen, biomass, carbon dioxide, impurity, MAB and water. Media, impurity and MAB are created as new components. The air is registered as a stock mixture by selecting task, stock mixture and register,edit/view properties. 2. The unit procedure is added. Select the unit procedures, batch vessel and fermentor to add the batch vessel procedure in a fermentor. 3. The streams are added by selecting the connect mode. There are 3 inlet streams; air, water and feed. The outlet streams are batch and carbon dioxide. 4. Right click on the fermentor to select operation data to add unit operations. The unit operations are charge 1, charge 2, heat, ferment, cool and transfer out. 5. The inlet water (charge 1) is set as 10 000 kg/batch while biomass and media feed are 50kg/batch and 450kg/batch respectively. 6. The properties of each unit operation are added on the operation condition. For charge 1, the setup duration is 5 minutes and the volume flow rate is set as 100L/min. For charge 2, the setup duration is 20 minutes and mass flow rate is 10kg/min. For heat, select the temperature, heating rate and setup time to set as 37C, 0.5C/min and 5 minutes. For the ferment, the operation time is 24 hours, heat transfer agent is chilled water and final temperature is 37C. Then select the vent/emission, thick on the emission, emission component is carbon dioxide and the vent stream is also carbon dioxide. Also, click the R to set substrate as media and rate reference components as biomass. For cool, the final temperature is 5C, cooling agent is Glycol and setup time is 5 hours. Lastly, for transfer out, the process time is 5 hours. 7. The process is saved and the analysis is complete by: click on the solve M&E balances on the main toolbar. The reports and charts can be selected on the reports and Gantt chart menu.


Process Flow Diagram (PFD)



1. Calculate the amount of air to be supplied for the aerobic fermentation process. 2. Determine the size of the fermenter necessary for this batch process. 3. What is the composition of vent & how much CO2 is produced in the batch process? 4. What is the composition of the broth and how much MAB is produced per batch? 5. How long is the heating and cooling operation required? 6. What is the total time required to complete one batch? If the total time per batch requires 36 hours, what is the maximum duration that the transfer out operation can prolong?
day h Complete Recipe P-1 in FR-101 CHARGE-1 CHARGE-2 HEAT-1 FERMENT-1 COOL-1 TRANSFER-OUT-1 CHARGE-1 (1.76 h) CHARGE-2 (1.17 h) HEAT-1 (0.48 h) FERMENT-1 (24.00 h) COOL-1 (1.15 h) TRANSFER-OUT-1 (5.25 h) 8 1 16 24 32 2 40 48 56 day h

Figure: Gantt chart showing the total time required. From the above chart, the total time required to complete one batch fermentation is 33.81 hours. Here, the transfer out duration is 5.25 hours.
day h Complete Recipe P-1 in FR-101 CHARGE-1 CHARGE-2 HEAT-1 FERMENT-1 COOL-1 TRANSFER-OUT-1 CHARGE-1 (1.76 h) CHARGE-2 (1.17 h) HEAT-1 (0.48 h) FERMENT-1 (24.00 h) COOL-1 (1.15 h) TRANSFER-OUT-1 (7.44 h) 8 1 16 24 32 2 40 48 56 day h

Figure: Gantt chart is the total required time is 36 hours

From the above chart, in order to get total required time of 36 hours, the operation data at the transfer out is changed. A total of 36 hours is get by selecting on the process time and set as 7.19 hours. Hence, the transfer out time is 7.44 hours.

7. Show graphically, when the biomass attain stationary phase by providing the concentration profiles for media, biomass and MAB. [Hint: Select the required variables under profiles tab of ferment operation to store data; To generate the data after simulation, right click on the fermentation procedure and select Dynamic Data Records \ FERMENT-1 \ Save in Excel Formatted File] Time (h) Biomass (g/L) 0 1.2 2.4 3.6 4.8 6 7.2 8.4 9.6 10.8 12 13.2 14.4 15.6 16.8 18 19.2 20.4 21.6 22.8 24 4.7171 5.9959 7.6213 9.6868 12.311 15.6369 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 16.6156 Concentration MAB (g/L) 0 0.0913 0.2074 0.3549 0.5423 0.7798 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 0.8497 media (g/L) 42.4542 37.8916 32.0921 24.7222 15.3591 3.4921 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Time vs concentration (biomass, MAB and media)

45 40 35 30 25 20 15 10 5 0 -5 0 5 10 15 20 25 30 concentration (g/L) concentration (g/L) concentration (g/L) Biomass MAB media

Graph: time versus concentration of biomass, MAB and media The graph above shows the concentration of biomass, MAB and media from initial time of 0 until 24 hours. The concentration of media is decreasing rapidly from 0 until 7.2 hours and reaches 0. Media is the reactant in the fermentation process, hence is being used for the production. The initial concentration is media is 42.4542 g/L. The concentration of biomass is increasing from the initial of 4.7171 g/L until 16.6156 g/L at t = 7.2 hours. Its concentration becomes constant at 16.6156 g/L until 24hours. The concentration of MAB is also increases but slightly from the initial time until 7.2 hours and constant at the value of 0.8497 g/L until 24 hours. The products (biomass and MAB) are constant because there is no more reactant (media) to used for the production.