Prática de Shadowing: Bioprocessing Part 1: Fermentation - Aprenda a falar inglês com vídeo
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we all know something about fermentation it's a process used countless times each day to make a variety of dairy products,
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baked goods, and beverages.
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We sometimes think of it as letting foods go bad, but in a controlled way.
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With a little help, milk becomes yogurt, bread rises, and grains decompose,
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creating alcoholic beverages and alternative fuels.
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But looking at these examples only gives us a clue as to what's really happening,
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and how we can use the power of fermentation to cost -effectively create a broad array of biological products. So, what is fermentation?
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A cell can be thought of as a microfactory.
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These cells can be bacteria, fungi, or specific cells from mammals, plants, or insects.
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In biotechnology, these cells are used to manufacture a product in a process called fermentation.
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For yogurt, buttermilk, and cheese, we use bacteria.
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To make breads and alcoholic beverages, we use yeast, a fungus.
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And the production of some vaccines require the growth of mammalian cells that are infected with a specific virus.
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The product the cells manufacture is usually a chemical the cells contain naturally,
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or a substance that the cells have been genetically altered to create, or even a metabolic waste product of the organism's growth,
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like one of our examples, alcohol.
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There are too many everyday products created by commercial scale fermentation to even list, but some common ones include amino acids,
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biopharmaceuticals, dyes, enzymes, food products, lipids, steroids, and vitamins.
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Fermentation is a reasonably simple process.
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A cell is selected based on its ability to produce the desired product.
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A seed stock of cells is put into a small amount of media.
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Media provides the nutritional products the cell needs to grow.
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When the population of cells has grown and consumed most of the nutrients, it's moved into a larger vessel with more growth media,
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and the process repeats.
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This scaling up is complete when the quantity of cells is large
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and healthy enough to transfer into a production vessel often referred to as a bioreactor
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or fermenter with plenty of fresh media now available
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and under tightly controlled conditions the cells grow and manufacture product
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when the fermentation is complete the product is harvested fermentation is known as an upstream biotechnology process
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it occurs early in the production flow before recovery, purification, formulation, filling, and packaging.
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To better understand the fermentation process, we should first find out a little bit about the cells we use
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and what they may require to reproduce and stay healthy.
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Different cells have different needs.
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Some are aerobic, they need oxygen, while others are anaerobic and do not require oxygen.
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All cells require nutrition.
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A properly formulated media contains the necessary nutrients to allow cells to grow and produce.
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The fermenter mixes the cells evenly throughout the media to suspend the cells and supply the oxygen necessary for growth.
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Effective and efficient fermentation requires rigorous monitoring and control of the environment within the bioreactor.
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Key factors include temperature, pressure, pH, which is a measure of how acidic or alkaline the media is,
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oxygen, usually measured as dissolved oxygen within the media, and nutrient levels.
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Although the environment and the media are tailored to the needs of specific cells, the life cycle of almost all batches follows a predictable pattern.
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The growth pattern has four phases, lag, exponential or log.
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stationary, and death.
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When a cell is first introduced to fresh media, it has to adapt to its new environment.
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This creates a lull or lag in the growth timeline.
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After the organism adapts, the batch takes off.
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The cells begin dividing at a constant rate, an exponential or logarithmic or log increase,
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doubling, then doubling again, and on and on.
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As the nutrients in the media are consumed, toxic metabolic waste products build up, cells begin to die, and growth slows.
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When it reaches the point that just as many cells are dying as are dividing, the batch enters the stationary phase.
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This is the point at which the key nutrients are completely consumed, the fermentation is stopped, and the fermented broth is harvested.
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If the fermentation were allowed to continue, the cells would enter the death phase.
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More cells die than divide.
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And, similar to the exponential phase, the death rate increases logarithmically.
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Now that we have a basic understanding of how fermentation works, let's look at an actual process and see how it all comes together.
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For our sample process, we will look at the production of Green Fluorescent Protein, or GFP.
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GFP is broadly used as a biological marker.
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It's a fluorescent dye that's very well tolerated by most cells and doesn't interfere with normal cellular function.
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In the GFP fermentation process, we'll need to add an antibiotic to protect the purity of the batch, and then, late in the process,
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a biochemical inducer to turn on the GFP gene.
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Our materials for this process will include a bacteria seed stock, in this case E coli,
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that has been genetically enhanced to produce GFP.
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The basic ingredients for a compatible media which include nutrients, stabilizers, an antibiotic, and an anti -foaming agent,
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and IPTG, which is the biochemical inducer that switches on the GFP gene.
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The equipment that we'll be using includes a 300 -liter bioreactor, a UV -Vis spectrophotometer to monitor the optical density,
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which is a measure of the concentration of cells in the bioreactor, a glucose analyzer to measure glucose,
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a key nutrient, an offline pH meter to help track the acid -base balance and adjust online measurements if needed,
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and a broth tank for our final product.
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The bioreactor is equipped with a water jacket around the vessel to regulate temperature and integrated sensors to monitor key environmental factors,
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including dissolved oxygen, pH, internal temperature, water jacket temperature, and vessel pressure.
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The reactor also has an agitator, dedicated ports for adding seed stock and media ingredients,
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separate ports for acid and base supplement, air filters for supply and exhaust,
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and valves for drawing samples and for harvesting.
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Most fermentation and monitoring functions can be managed from the bioreactor's dedicated process controller.
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Before the fermentation process can begin, the area must be prepared.
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Preparation includes removing equipment and material that won't be used in the process, cleaning and sanitizing the area and equipment,
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and sterilizing equipment as required by the SOPs, Standard Operating Procedures.
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Sterilization is used to eliminate unwanted microorganisms, which can grow naturally in the fermentation media and process equipment.
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Also, all required materials and documentation should be gathered and prepared.
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And all process control software should be loaded and verified.
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The fermentation batch process will be guided and documented with the BPR, Batch Process Record.
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The batch record leads the operator through the process, step by step, with each step requiring a sign -off and separate verification.
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This record also includes spaces for documenting key times, activities, and instrument readings.
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The GFP fermentation process really begins with the expansion of our bacteria seed stock.
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After removing the specially modified E coli from the freezer and thawing it,
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it's used to inoculate a small amount of fresh media in a shaker flask.
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After the number of cells has reached the target amount, the thriving cells are ready for fermentation.
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Meanwhile, in the fermentation area, operators begin with a complete check of all critical equipment.
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Valves, caps, and lines are checked.
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Hoses are tightened.
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Probes are verified and calibrated.
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and 10 kilograms of HPW is added to the vessel.
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The bioreactor is brought up to normal process pressure and held there in order to check for leaks.
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The pressure is monitored over a 30 -minute period.
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If a leak is detected, the problem is corrected and the test is run again.
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Once the reactor passes the test, we are ready to mix the media in the vessel.
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The agitator is turned on, and the ingredients are added.
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Yeast extract, triptych soy broth,
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ammonium chloride, sodium biphosphate, monopotassium phosphate,
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and an anti -foam compound.
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Once all the initial ingredients are in, another 10 kilograms of high -purity water is added.
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All ports and valves are closed.
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All condensate valves are opened.
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And the bioreactor begins an SIP, sterilize -in -place cycle.
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The target for sterilization is 121 degrees Celsius for 30 minutes.
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As soon as the temperature climbs to the targeted temperature, the condensate valves are closed, and the SIP cycle completes automatically.
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Both the vessel and the media are now sterile, and we're ready to add the final ingredients to our media.
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The glucose hose is attached to the vessel, the connection is steamed to sterilize it,
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and the separately sterilized glucose antibiotic solution is pumped into the vessel.
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Then a manual pH reading of the media is taken and the bioreactor is set up for its fermentation cycle.
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After the inoculation hose is connected to the reactor and steamed for 20 minutes,
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the expanded seed stock is pumped into the reactor containing the media.
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Fermentation now begins.
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The operator takes zero -hour readings and begins to regularly monitor batch temperature,
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agitator RPMs, dissolved oxygen levels, pH, vessel pressure, optical density,
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airflow rate, and glucose concentrations.
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Optical densities and glucose concentrations are of particular interest, so they're graphed as well as documented.
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When the targeted levels of glucose and optical density are achieved, it's time to add IPTG to the vessel to activate
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or turn on the expression of the green fluorescent protein in the cells.
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After allowing enough time for the cells to produce green fluorescent protein usually five hours more final readings are taken
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and a sample is drawn to check the percentage of cell
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solids the product is now referred to as broth the broth
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which contains spent media and cells is complete
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when the key nutrient glucose is mostly consumed and the batch has reached the desired concentration.
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The batch is then cooled down, pumped into a broth tank, and labeled with the batch number, volume, time, and date.
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The fermentation process is now complete.
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The harvested broth will now move downstream to the recovery process, where the cells will be ruptured to free the green fluorescent protein,
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and the protein will be separated from the other broth components.
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Vocabulário e dicas de fala para esta lição
Esta aula de conversação usa o vídeo “Bioprocessing Part 1: Fermentation”. As palavras que mais se repetem: cell, fermentation, process, product, vessel. Este vídeo tem 128 frases e 1692 palavras para praticar shadowing. A fala dura 14:32. O falante mantém um ritmo estável de cerca de 116 palavras por minuto, confortável para o shadowing. Apenas 72% das palavras estão entre as 3.000 mais comuns do inglês, por isso o vocabulário é exigente.
Vocabulário principal deste vídeo
15 palavras do vídeo que vale a pena aprender, com pronúncia e significado:
| Palavra | Pronúncia | Significado |
|---|---|---|
| vessel substantivo | /ˈvɛsəl/ | barco, navio |
| batch substantivo | /bæt͡ʃ/ | fornada |
| glucose substantivo | /ˈɡluː.kəʊz/ | glicose, glucose |
| nutrient substantivo | /ˈnuː.tɹi.ənt/ | nutriente |
| oxygen substantivo | /ˈɑksɪd͡ʒən/ | oxigênio, oxigénio |
| monitor substantivo | /ˈmɑ.nɪ.tɚ/ | monitor, observador |
| valve substantivo | /ˈvælv/ | válvula |
| ingredient substantivo | /ɪnˈɡɹiːdi.ənt/ | ingrediente |
| density substantivo | /ˈdɛn.sə.ti/ | densidade |
| concentration substantivo | /ˌkɑn.sənˈtɹeɪ.ʃən/ | concentração |
| harvest substantivo | /ˈhɑɹ.vəst/ | colheita |
| consume verbo | /kənˈsum/ | consumir |
| reactor substantivo | /ɹiˈæktɚ/ | reator |
| acid substantivo | /ˈæsɪd/ | ácido |
| operator substantivo | /ˈɑpəˌɹeɪtɚ/ | telefonista |
Phrasal verbs que você vai ouvir
| Palavra | Pronúncia | Significado |
|---|---|---|
| cool down verbo | /kuːl ˈdaʊn/ | esfriar, arrefecer |
| find out verbo | descobrir | |
| sign off verbo | desconectar, autorizar | |
| take off verbo | tirar |
Gramática neste vídeo
As estruturas que o falante mais usa, com as palavras exatas do vídeo:
| Estrutura | No vídeo |
|---|---|
| Voz passiva be + particípio passado — o foco está no que acontece, não em quem faz | can be thought · are infected · have been genetically altered |
| Present perfect have/has + particípio passado — uma ação passada que ainda importa agora | have been · has grown · has been |
| Orações relativas who / which + oração — informação extra sobre uma pessoa ou coisa | IPTG, which is · microorganisms, which can |
Pronúncia para ficar de olho
O falante usa 6 contrações e formas reduzidas, como we'll, doesn't, they're. Diga-as na forma curta, do jeito que você ouve.
- Os sons de “th”: gather /ˈɡæðɚ/, thrive /θɹajv/, broth /bɹɔθ/
- Os sons de “sh” e “zh”: concentration /ˌkɑn.sənˈtɹeɪ.ʃən/, stationary /ˈsteɪʃəˌnɛɹi/, efficient /ɪˈfɪʃənt/, preparation /ˌpɹɛpəˈɹeɪʃ(ə)n/, nutrition /njuːˈtɹɪʃ.ən/
- Palavras longas — acerte a sílaba tônica: ingredient /ɪnˈɡɹiːdi.ənt/, concentration /ˌkɑn.sənˈtɹeɪ.ʃən/, operator /ˈɑpəˌɹeɪtɚ/, antibiotic /ˌæn.taɪ.baɪˈɑ.tɪk/, naturally /ˈnæt͡ʃ.(ə.)ɹə.li/
Sons difíceis para falantes de português:
- Consoante final — sem acrescentar um “i” depois: nutrient /ˈnuː.tɹi.ənt/, ingredient /ɪnˈɡɹiːdi.ənt/, harvest /ˈhɑɹ.vəst/, acid /ˈæsɪd/, pump /pʌmp/
- /l/ final — a língua toca o céu da boca, não vira “u”: vessel /ˈvɛsəl/, optical /ˈɑptɪkəl/, biological /ˌbaɪ.əˈlɑ.d͡ʒɪ.kəl/, meanwhile /ˈmiːnwaɪl/, label /ˈleɪ.bəl/
- /h/ — um sopro suave, diferente do “r”: harvest /ˈhɑɹ.vəst/, alcoholic /ˌæl.kəˈhɔ.lɪk/, hose /hoʊz/, enhance /ɪnˈhæns/
Como praticar com este vídeo
- Ouça o vídeo inteiro uma vez sem falar e anote as palavras que você não conhece.
- Faça shadowing frase por frase na velocidade normal, repetindo cada uma até o seu ritmo ficar igual ao do falante.
- Grave a sua voz e compare com o original, prestando atenção a palavras como vessel, batch, glucose.
O que é a Técnica de Shadowing?
Shadowing é uma técnica de aprendizado de idiomas com base científica, originalmente desenvolvida para o treinamento de intérpretes profissionais. O método é simples, mas poderoso: você ouve áudio em inglês nativo e repete imediatamente em voz alta — como uma sombra seguindo o falante com 1-2 segundos de atraso. Pesquisas mostram melhora significativa na precisão da pronúncia, entonação, ritmo, sons conectados, compreensão auditiva e fluência na fala.