We all know something about fermentation. It’s a process used countless times each day to make a variety of dairy products, baked goods and b__s. We sometimes think of it as / letting foods go bad / in a controlled way / little help / milk becomes y__ / bread rises, and grains decompose, creating alcoholic beverages and alternative fuels. But looking at these examples only gives us / clue / as to what’s really happening / how we can use the power of fermentation to cost-effectively / create a broad array of biological products.
So, what is fermentation?
As cells can be thought of as / micro / or specific cells from m__s, plants, or insects. In biotechnology, these cells are used to manufacture a product in a process called fermentation. For y__, butter and cheese, we use bacteria. To make breads and al__ b__s, we use y__, a f__. / production of some v__s require / m__ cells that / specific virus. The product / manu__ / is usually a chemical the cells contain naturally, or a substance that the cells / genetically altered to create, or / waste product / organism’s growth, like one of our examples, al__.
there are / everyday / commercial-scale fermentation / even / list / but some common ones include a__, bio__, __, __, / food products, __s, s__ and vitamins.
Fermentation is a reasonably simple process. __ __ __ selected / the desired product. A seed stock of cells is put / small amount of media / provides / nutritional products the cell needs to grow. When / population / and consumed most of the nutrients, it’s moved into a larger vessel / media / and the process repeats. This scaling-up is complete when the quantity of cells is large and healthy enough / into a production vessel, often referred to as a ‘bioreactor’ or a ‘fermenter’. With plenty of fresh media now available / under / tightly controlled / product / when the fermentation is complete, the product is harvested. Fermentation is known as an upstream / biotechnology process; It occurs / production / before / purification, formulation, filling and packaging.
To better understand the fermentation process, we should first find out a little bit about / what they may require to reproduce and stay healthy.
Different cells have different needs. Some are a__bic; They oxygen / while / do not require oxygen. All cells require nutrition / a properly formulated / media / to allow cells to grow and produce. The fermenter mix__ / cells evenly throughout / oxygen necessary for growth.
Effective and efficient / r__ monitoring / environment within the bioreactor. Key factors / pressure / which / how / the media is / oxygen / usually measured / within the media, and nutrient levels. Although the environment / needs / follows a predictable pattern. The growth pattern has four phases;/ exponential / or log / stationary and death.
When a cell / first introduced / fresh media / new environment / this / or / in the growth timeline. After the organism a__s, the batch takes off. The / dividing / at a constant rate / exponential / logarithmic / doubling, then doubling again, and on, and on. As the nutrients in the media are consumed, toxic / cells begin to die / growth slows. When it reaches __ point / just as many cells are dying / the batch enters the stationary phase. This is the point at which the key nutrients are completely consumed / the fermentation / stopped / and / broth is harvested. If the fermentation / allowed to continue / enter the death phase / than / and, similar to the exponential phase, the death rate increases logarithmically.
Now that we have a basic understanding of how fermentation works, / look / actual / all comes together. For our sample process, we will look at / or GFP / GFP / biological marker. It’s a f__ d__ / very well tolerated by most cells. It doesn’t interfere / function / in the GFP fermentation process, we __ need to / to protect / purity / in the process / to turn on the GFP gene. Our materials for this process will include a bacteria seed stock / in this case, e-__, that has been genetically / enhanced to produce / basic / compatible media, which include nutrients, stabilizers and anti / anti-__ agent / which is the biochemical __ that switches on the GFP gene. The equipment that we’ll be using includes / bioreactor / to monitor the __ density, which is the measure the concentration of cells in the bioreactor / g__ / measure / a key nutrient / An offline pH meter to help track the acid-base balance / if needed / for our final product.
The bioreactor is equipped with a water-jacket around / to regulate temperature. / to monitor / environmental / factors / temperature, water jacket temperature, and vessel pressure. The reactor also has an agitator __ __ __ adding seed stock in media ingredients / ingredients / acid / base supplement / air filters for supply and exhaust / and / samples / and for harvesting / most fermentation __ monitoring functions can be managed / dedicated process controller. Before the fermentation process can begin, the area must be prepared. Preparation includes removing / that won’t be used in the process / cleaning and s__ing the area and __ equipment / and sterilizing as required / standard operating procedures. S__ / procedures / eliminate unwanted microorganisms which can grow naturally in / fermentation / media / equipment. Also, all required / documentation / should be gathered prepared / and all process control software should be loaded and verified.
The / batch / guided / with / BPR: Batch Process Record__ The batch record leads the operator through the process, step by step / requiring / certain / verification. / This record also includes spaces for documenting key times / activities / and instrument readings.
The GFP fermentation process really begins with the expansion of __ bacteria seed stock. After removing / modified e-__ / freezer / it’s used to i__ __ small amount of fresh media in a shaker flask. After the number of cells / target / are ready for fermentation. Meanwhile, in the fermentation area, operators begin with / complete / all critical equipment / valves, __s and lines are checked. Hoses are tightened. Probes are / verified / and ten kilograms of HPW, high-purity water, \ is added to the vessel. The / normal process pressure __ __ __ in order to check for leaks. / The pressure is monitored over a 30-minute period / the test is run again.
Once the reactor passes the test, we are ready to mix the media in the vessel. / the agitator is turned on / and the ingredients are added. / y__ extract, __ __ s__ broth, __ chloride, sodium b__, m__ and __ __ __ compound. Once all the initial ingredients are in, another ten kilograms of / high-purity water is added. All ports and valves are closed. All __ valves are opened, and the bioreactor begins / SIP / cycle. The target for s__tion / one / degrees / for 30 minutes. As soon as the temperature / targeted temperature / are closed, and / SIP / cycle / completes automatically. Both the vessel / are now sterile / final ingredients to our media / the g__ hose is attached to the vessel / sterilize it, and / solution is pumped into the vessel. / then, a manual / media is taken. And the bioreactor is set up for its fermentation cycle. After / hose / connected to the reactor / steamed / for 20 minutes / expanded / into / containing the media.
Fermentation now begins. The operator / regularly / dissolved oxygen levels / pH / vessel pressure / density / air flow rate / concentrations / concentrations are / particular interest / as well as / documented / targeted / are achieved, It’s time to / add / to / activate / turn on / expression / cells. After allowing enough time / cells / usually / five hours more, / final readings are taken. / sample /
The product / now referred to as ‘broth’. The broth / contains / spent / complete / mostly consumed / desired concentration / down / and labeled / time and date / fermentation / now complete. The harvested broth / now / downstream / recovery process / free / and / be separated from the other broth components.
