1. F215 5.2.2 Biotechnology By Ms Cullen

2. What is biotechnology? Can also be known as biotech. Refers to any technological or industrial use of organisms (or components of organisms), to produce products or processes for a specific use. Examples: medicine agriculture food processing aquaculture non-food industrial processes environment

3. Medicine Pharmaceutical production Human proteins Antibiotics Synthetic drugs Diagnostic testing Antibody testing Imaging Gene profiling Vaccine development Traditional vaccines Recombinant vaccines Gene Therapy Vectors for gene therapy Gene delivery systems Organ Transplants Organ transplant Xenotransplant Stem cell technology

4. Food Production and Processing Agriculture & Aquaculture Inherited disease resistance Herbicide / insecticide resistance Increased nutritional value Improved stress tolerance Increased production rate Food Processing Fermentation technology Enzymatic modification Improved process control and efficiency Enhanced characteristics (eg flavour, shelf life, nutritional value) Diagnostic food testing

5. Environment Bioremediation Organic spills (petroleum) Metal contamination Radioactive material Biosensors Chemical monitoring Pathogen monitoring Water treatment Waste water sewage Pest control Weed control Control of feral animals

6. Industry Biofuels Production of fuels using alternatives to fossil fuels (eg sugar cane, maize) Biocatalysts Synthesis of chemicals Brewing industry Pharmaceutical production Green plastics Production of plastics using petroleum alternatives (eg sugar cane) Mining Ore leaching Metal extraction Coal cleaning

7. Why are microorganisms used in biotechnology?

9. Activity 20 Investigating population growth

10. The standard growth curve for a culture of microorganisms

11. Questions: 1.Explain why there is an initial lag in the growth of a microorganism placed into a new culture. 2.Suggest how this lag can be reduced when starting a new culture. 3.Describe the effect on microbial growth of adding fresh nutrients and removing toxic by- products.

12. Primary metabolites During the log phase many intermediate metabolic products are produced, which are needed by the microbes for growth or to provide energy. These are known as primary metabolites and are produced in excess and will accumulate in the culture. They can then be extracted and purified. Primary metabolites form at same rate as the cells grow, therefore their growth curves are very similar. Example: production of ethanol from yeast

13. Secondary metabolites These are organic compounds, but they are not necessarily essential for survival of the microorganisms. They are often produced as defence mechanisms or to allow the organism to compete with another. Many secondary metabolites have beneficial uses to humans eg morphine, atropine, penicillin Secondary metabolites are produced after the active phase of growth has ended and the culture is in the stationary phase. A limited number of microorganisms produce secondary metabolites, when 1 or more nutrient is depleted from the growth medium. Many important antibiotics are produced as secondary metabolites from bacteria and fungi.

14. Growth and product curves showing the production of (a) a primary metabolite and (b) a secondary metabolite

15. Question: 1.Explain why the curve for a primary metabolite production closely resembles the microorganisms growth curve. 2.Explain why this is not the case for production of a secondary metabolite. 3.Explain the differences between a primary and a secondary metabolite in microbial culture.

16. Generalised diagram showing the features of a large-scale industrial fermenter

17. Fermenter Read through biofact sheet 47 on fermentation http://videos.howstuffworks.com/science-channel/29783-100-greatest-discoveries-penicillin-video.htm

18. Asepsis This is the practice of preventing contamination of cultures by unwanted microorganisms. Q Why is this important?

19. Q What aseptic techniques can be used to prevent contamination?

20. Industrial Enzymes

21. Enzymes Enzymes are important in commercial and industrial processes as they accelerate chemical reactions to produce a useful product or effect. Some enzymes used remain in their cells, others are extracted and purified. In each case if the enzymes are trapped within an insoluble agent their efficiency is increased. This is called enzyme immobilisation.

22. Enzymes and their uses EnzymeSubstrateUses ProteasesProteinWashing powder, leather production, treatment of blood clots AmylasesCarbohydratesSweetners LipasesLipidsWashing powder CellulasesCelluloseFabric conditioners, fruit juice production – prevents cloudiness LactasesLactoseProducing lactose free milk PectinasespectinFruit juice production

23. Enzymes Enzymes are sourced from a variety of microorganisms, bacteria, fungi and yeast. The bacteria Bacillus and fungi Aspergillus are particularly useful. Microorganisms can be genetically modified to increase the yield of enzymes they produce. The enzymes are purified by grinding cells and/or adding alkalis. The cell debris is then removed by filtration and centrifugation. The enzymes are then precipitated using ammonium sulphate. Further techniques may involve techniques such as filtration, chromatography and electrophoresis.

24. Immobilising enzymes Enzymes are expensive and therefore being able to recycle them is a cheaper option. The best way to do this is by immobilising them. 1.An enzyme solution is mixed with a solution of sodium alginate. 2.Little droplets of this mixture are added to a solution of calcium chloride. 3.The sodium alginate and calcium chloride react to form a jelly, which turns the droplets into little beads. 4.The enzyme is held in the bead and is therefore immobilised.

25. Now have a go yourself! Complete steps 1-6 of Activity 22 - Investigating Immobilised Pectinase

26. Whilst you are waiting: Using P.164 -5 in OCR A2 Biology make notes on: - the advantages and disadvantages of using immobilised enzymes - methods for immobilising enzymes

27. Now complete Activity 22 and graph your results Complete Qs for Activity 22

28. Activity 23 – Investigating Immobilised Lipase Draw a graph of your results and answer Qs