Team:ECUST China/Hardware
Our hardware carries on the innovation in term of recycling yeast slurry after beer fermentation to gain pure phycocyanin and at the same time produce phycocyanin beer to maximize the profits and reduce the environmental pollution that may cause from the extraction step to produce phycocyanin from algae.
Before the building of the factory we should determine the process of our fermentation and draw the fermentation process down. After referring to the literature reviews or books as‘Principles of Chemical Engineering’, combining the characteristic of our yeast and consulting to the professor specialized in fermentation, we finally determined our Phycocyanin Beer Production Factory as follows:
The whole process is that we introduce our modified yeast, filtered air and fermented related compounding to the pure culture tank, fermented for 3 hours until the OD index of our yeast turn to approximately 1.4. (This means our yeast has been growth to the primary state of logarithmic phase, fully prepared for the inducing process followed.）Then the primary fermented broth will be introduced into the secondary seed fermenter, further enlarging the fermentation. And in the primary fermentation step in the seed fermenter we will add galactose artificially whose ratio is 2%. (Considering that our expression gene are big molecules, adding the inducer at the logarithmic phase can largely reduce the impact on yeast growth. ) After fermentation, the yeast fermentation broth will be introduced into centrifugal machine and then plate-and-frame filter press to be separated to gain fresh yeast, preparing for the following beer fermentation.
Then in beer fermentation process, malt, water and hops will be introduced separately into pulverizer, gelantinization tank, saccharifying tank, filter tank, boiling pot, cyclotron precipitation, cyclotron precipitation, cooling bath and then anaerobic fermentation tank. The detailed process is follows. Large malt 25kg, small malt 15kg are added to the pulverizer to crush, then they will be introduced to the make-up tank for primary infiltration and heating. Then in the gelantinization tank, heating it to 52 degrees (the surface temperature is 2 degrees lower than the actual temperature) ,adding 40g CaCl2 and 30ml lactic acid to adjust the pH to 5.3-5.5. The process of saccharifying contains 52℃ stirring for 30 min, 65℃ heating for 70 min, 72℃ heating for 20 min and then filter and stand for 20 min to measure the saccharifying degree. Then the ingredients will further introduced to the boiling pot to boil for 70 min, then adding 50g hops for boiling 10mins, adding 80g hops for boiling 30mins, adding 45g hops for boiling 60mins. They were driven into the rotary sedimentation tank standing for 20 min and then fed into the anaerobic fermentation tank.The initial fermentation temperature is controlled at 18 ℃. After 24 hours, put the resin to precipitate and ensure the sugar content below 5.0. Then raise the temperature higher for 20 ℃ and boost the pressure, using reduce diacetyl for 2 days and then lower the temperature to 5 ℃( 0.3 ℃ per hour).Add Our modified fresh yeast, continue to cool down to 1 ℃ (0.1 ℃ per hour) and then enter the post fermentation. Ferment for a week and then finally produce phycocyanin beer through the filter tank, depending on the pH regulation among 4.3-4.5. The filter is important to ensure the taste of the beer for too much yeast existing in the beer will give an awful smell, while there still existing an problem that our phycocyanin was expressed in the cytosol of the yeast. How can we get the phycocyanin even after the filter? Learning that there is a process called self autolysis which will release intracelluar molecules in the last step of fermentation, we let the beer turn blue through balance the degree of self autolysis and the taste of the beer. What’s more, after fermentation, we not only get our nutrient phycocyanin beer but also get phycocyanin yeast slurry which can be further processed to produce large quantity of pure phycocyanin, making our factory economically valuable.
The constructing of the 3D hardware can guide us to construct our idea of how to implement our project really and making the project possessing integrity. You can set about building your fermentation factory using CAD, 3D MAX or other 3D-modeling software. We learn it by ourselves and through the guidance of professional person we finally construct our factory as follows:
To build the 3D model of a factory, you not only have to know the process flow diagram but also have to know the details and features of the specific tanks. We set the volume of our anaerobic beer fermentation tank of 400L and the saccharifying tank, boiling pot of 200L, meaning that two period of saccharification can fill one tank of fermenter. In the yeast producing part, we set our secondary seed fermenter’s volume of 400L and make-up tank of 200L, also two period one fermenter. Through controlling the quantity of supplementary fresh yeast, we can well control the operation of our whole fermentation factory. You can see the detail of our factory as follows.
To test whether our hardware is theoretical feasible, we have to know the approximate economic cost
of it. What we calculated is the total ingredient’s cost of our factory such as the hops, malt, air
and all the other materials the factory called for. The costing of the factory itself various a lot
and it’s economic effect can be decrease along with the using time. And the data was reasonable to
be used to compare with other factories.
The calculation process is follows:
The required yeast content is calculated at 0.5% of the total volume of beer tank.
Assume that the final product unit is，the fermentation unit is
Daily tank volume is determined by annual output:
(one
process one day)
Therefore, a total of 6 primary seed tanks are required.
Secondary seed fermentation:(assume that
the loss ratio is 15%)
Primary seed tank:
The auxiliary time is about 10 hours:
Having the volume of all the materials we need in the first and second step fermentation , we now can calculate the cost of the total process. One process of the fermentation period calls for 10 days and it calls for raw materials: large malt 25kg, small malt 15kg,hops 0.175kg, yeast culture media 9.1*10^(-7)kg, thinner feed 6*10^(-7)kg, complete feed 1.11*10^(-6)kg, creosote oil 1.2*10^(-8)kg, ammonia3*10^(-8)kg. (Note that the fermentation of the bear calls for only 0.5% of the total volume of beer while one tank of yeast’s fermenter can produce large quantity of yeast, so the order of magnitudes for yeast fermentation is small.While you can also produce more fresh yeast to sell it separately which is also a good idea.) Finally the annual cost of the materials of our factory is about 3750 dollar per year, giving birth to 14600L phycocyanin-beer per year.
(Our calculation just point out a way to count it and give some useful data, Their are more details should be added to the cost, such as the water, electric charge and other administration fees all should be taken into consideration.)
Based on this hardware, we can later magnify the production scale with increased size and parallel number of bioreactors. The factory is characterized with our own yeast's feature and can really make the 'Magic Blue' come true!
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+86 15821261548
IGEM_ECUST@outlook.com
ID:IGEM_ECUST
ECUST_China
East China University of Science and Technology
Shanghai,China
