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Classical Biotechnology: FERMENTATION


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Classical Biotechnology: FERMENTATION

  2. 2. recall: biotechnology •ancient •early history as related to food and shelter, including domestication ! •traditional (classical) •built on ancient biotechnology •fermentation promoted food production and medicine ! •modern •manipulates genetic information in organism •genetic engineering
  3. 3. Grabbed from the PPT lectures of Professor/Dr. Arnold V. Hallare, (2013)
  4. 4. what to learn today… •overview of metabolism: aerobic and anaerobic respiration •Fermentation in Plants •Fermentation in Animals •Fermentation in Humans •traditional biotechnology: fermentation •virtual lab
  5. 5. recall: metabolism
  6. 6. cellular respiration: a catabolic reaction •process of making ATP by breaking down organic compounds •exergonic •oxygen (O2) requiring •uses energy extracted from macromolecules (glucose) to produce energy (ATP) and water (H2O) enzymes 6O2 + C6H12O6 6CO2 + 6H2O oxygen glucose carbon dioxide water ADP + Pi ENERGY transfer between enzymes, other molecules ATP
  7. 7. stages of aerobic respiration • glycolysis: cytosol • krebs cycle: mitochondrial matrix • ETC: inner mitochondrial membrane You may watch a video here about GLYCOLYSIS: http://
  8. 8. anaerobic respiration (fermentation): if oxygen is absent Glucose Pyruvic acid
  9. 9. organic acids instead of atp
  10. 10. prokaryotes vs eukaryotes
  11. 11. anaerobic respiration in plants In response to flooding stress
  12. 12. anaerobic respiration in animals slow twitch versus fast twitch muscles
  13. 13. slow twitch and aerobic respiration • example: dark leg meat of chicken • Specialised for slow, sustained contractions over a long period for endurance • contain lots of myoglobin which acts as a store of oxygen • Respire aerobically
  14. 14. slow twitch works best in: or if you wanna try duathlon when you try running the bdm ultramarathon (160km)
  15. 15. lactic acid in meat? • fast twitch • example: pectoral flight muscles (chicken breast) • for producing rapid, intense contractions of short duration for rapid movement • do not have myoglobin so Respire anaerobically • can accumulate lactic acid and leads to fatigue
  16. 16. fast twitch works best in: Provide the muscle power for rapid, fast movement e.g. a cheetah's burst of speed to catch a gazelle, or the gazelles burst of speed to escape the cheetah or to power up usain bolt’s legs in sprints
  17. 17. fermentation in ruminants
  18. 18. fermentation in humans •farting •When carbon dioxide is used as an electron acceptor, the product is either methane or acetic acid •Methane produced in our gut is produced by this process
  19. 19. fermentation in microbes
  20. 20. fermentation : classical biotechnology •the use of microbes to enhance food flavor •the use of microbes to manufacture of beverages •the use of microbes to make the dough rise
  21. 21. products of fermentation: beer •An alcoholic beverage produced by the fermentation of sugar-rich extracts derived from cereal grains or other starchy materials •ancient biotechnology: beer brewing •Sumaria (4000 BC) •Sikaru •Egypt (3000 BC) •Zythum •India (2000 BC) •Sura •China (2000 BC) •Kiu
  22. 22. yeast in beer brewing •1680 Antonie van Leeuwenhoek Observed yeast in beer •1837 - Cagniard Latour decsribed that Microbe is responsible for alcoholic fermentation •1866 - Louis Pasteur stated that Yeast was responsible for alcoholic fermentation •1883 - Emil Christian Hansen Developed pure culture technique and Isolated pure cultures of brewing yeasts Weiss Ale Lager Lab Kindly provided by Tom Pugh and David Ryder of Miller Brewing Company Kindly provided by Tom Pugh and David Ryder of Miller Brewing Company
  23. 23. microbes and beer brewing •malted barley Provides fermentable sugars, flavor, and color •hops Provides aroma and bitterness The Brewing Process Step Purpose Brewhouse Fermentation Lagering Starch Sugars Wort production Sugars Ethanol Flavor production Carbonation Flavor maturation Kindly provided by Tom Pugh and David Ryder of Miller Brewing Company The Brewing Process Malt Mill Mash Tun Cereal Cooker Lauter Tun Brew Kettle Hot Wort Receiver Wort Cooler Brink Fermentation Aeration Lagering Hops Kindly provided by Tom Pugh and David Ryder of Miller Brewing Company
  24. 24. microbes and beer brewing Yeast Metabolism During Fermentation Sugars Oxygen Glucose Pyruvate TCA Cycle Amino Acids Energy CO2 Ethanol Acetaldehyde Organic Acids Unsaturated Fatty Acids Sterols Amino Acids Esters Higher Alcohols VDK Sulfur Volatiles Membranes Kindly provided by Tom Pugh and David Ryder of Miller Brewing Company
  25. 25. products of fermentation: cheese •cheese are ripened curds •milk is treated with lactic acid bacteria and rennin to coagulate proteins •curds + whey = milk •different microbes in the early and late stages of processing of cheese = different cheese characteristics
  26. 26. types oTfy pcesh oef Cehseeese Acid Coagulated Fresh Cheese (lactic acid from bacteria) • no enzyme is used to finish the curd • Cottage and Cream Cheese Heat-Acid Precipitated Cheese (acid and heat precipitate/coagulate the protein and cause milk fat to curdle) • Add low amounts of acid to 75-100oC temp milk • High moisture and protein • Ricotta (Italy) Channa and Paneer (India) science of cooking
  27. 27. types oTfy pcesh oef Cehseeese Semi-hard Washed Cheese (washing cheese removes acid and lactose) • Acid and enzyme induced curdling • But removal of milk sugar and acid results in no fermentation results in a moist and less finished cheese • Gouda, colby, muenster, mozzarella … Hard Cheese (Low and High Temp) • Low moister makes a more dense hard cheese • Elevated temps and pressing drive off water • Cheddar, Romano, Parmesan, Swiss, science of cooking
  28. 28. swiss cheese and propionibacterium science of cooking Finishing Microbes Holy Cheese (cow)? – Propionibacteria: • Convert lactic acid to propionoic and acetic acid plus acetic acid and CO2. Also other flavors • Used to make Swiss Cheese • Need higher temps and time for bacteria to grow and produce • Growth requirements reflect origins of bacteria animal skin Lactic acid Propionoic acid + Acetic acid Carbon Dioxide (g) Finishing up…
  29. 29. Blue Cheeses – Based on Origen fungi and blue cheese Roquefort - France Cambreles- Spain Stilton- England Danish Blue Cheese Gorgonzola- Italy science of cooking Penicillium roqueforti and P. camberti BLUE = MYCELIA/ or growth filaments
  30. 30. making the cheese Non Starter– ripening Starter – acid producing 0 50 100 150 200 science of cooking Bacteria Growth Time (Days)
  31. 31. product of biotechnology: breads •biotechnology’s first utilization of microbes = bread making •Around 4000 BC, Egyptians used the living organism yeast to make bread •Airborne wild yeast accidentally got their bread dough, causing it to rise
  32. 32. the sourdough bread •microbe one (AEROBIC): yeast •makes carbon dioxide and bread will rise •microbe two: anaerobic: lactic acid bacteria •make lactic acid and acetic acid that give rich complexity of flavors
  33. 33. products of fermentation: wine •after bread comes wine: 3000 bc •converts sugars in grapes into alcohol
  34. 34. making your wine 1. harvest 2. processing 3. fermentation
  35. 35. making your wine 4. maturation 6. bottling & corking 5. fining and filtration
  36. 36. products of fermentation: yoghurt •FERMENTED MILK RESULTING TO A SEMI-SOLID CURD •LACTIC ACID BACTERIA = PROBIOTICS •AIDS IN DIGESTION •ACID PRODUCED DURING FERMENTATION CAUSES THE PROTEIN TO COAGULATE •Lactococcus lactis, Streptococcus thermophilus and Lactobacillus bulgaricus
  37. 37. how to make yoghurt Making Yogurt in 4 Simple Steps 1. Start with Cow, Sheep, or Goat milk. 2. Heat milk to 80 °C. Two purposes: • destroy existing bacteria • condition the proteins = begins the denaturing process (a whey protein molecule binds to a casein molecule which disrupts the casein bundles allowing them to make short branched micelle chains) 3. Cool milk to 40 °C and innoculate with bacteria 4. Incubate at 30 °C to 45 °C Casein before heat pre-treatment: Casein after heat pre-treatment: Casein after acid:
  38. 38. bacteria in yoghurt Milk Yogurt Casein protein micelles Bacteria produce acid (bundles) 10-7 meters in diameter Fat globule Acid causes Casein bundles to fall apart into separate casein molecules. These rebind to each other in a network that traps water. = makes a gel