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F215 control, genomes and environment

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Presentation on theme: "F215 control, genomes and environment"— Presentation transcript:

1 F215 control, genomes and environment
Biotechnology F215 control, genomes and environment Module 2 – Biotechnology and gene technologies

2 Learning Outcomes State that biotechnology is the industrial use of living organisms (or parts of living organisms) to produce food, drugs or other products. Explain why micro organisms are often used in biotechnological processes.

3 Biotechnology Biotechnology is the industrial use of living organisms to produce food, drugs and other products. Biotechnology has four major applications that affect our lives Healthcare and medical processes Agriculture Industry Food science

4 Using Micro organisms Features of micro organisms that make them suitable for large-scale industrial processes Rapid life cycles Reproduce asexually Very specific and simple requirements for growth Can be grown on waste materials from industry Does not raise ethical questions Bacteria have a single copy of each gene Simple control of gene expressions Wide range of metabolic pathways Some evolved to survive at high temperatures

5 Learning Outcomes Describe, with the aid of diagrams, and explain the standard growth curve of a microorganism in a closed culture.

6 Standard Growth curve

7 Growth curve in a closed culture
Lag phase Bacteria adjusting to new conditions Takes a while for enzyme production Log phase Number of bacteria increase rapidly Stationary Phase Rate of growth is equal to rate of death Decline Phase Death rate is greater than “birth rate” The first three stages represent a sigmoid growth curve

8 Learning Outcomes Explain the importance of manipulating the growing conditions in a fermentation vessel in order to maximise the yield of product required.

9 Large-Scale production
Microorganisms are cultured in large containers called fermenters The growing conditions within the fermenter are manipulated and controlled Precise growing conditions Temperature Type and time of the addition of the nutrient Oxygen concentration pH

10 A batch fermenter

11 Large scale production
Three examples are The production of penicillin The production of protease enzymes The production of mycoprotein

12 Learning Outcomes Compare and contrast the processes of continuous culture and batch culture. Describe the differences between primary and secondary metabolites.

13 Metabolism and metabolites
Metabolism (process) Sum total of all the chemical reactions Processes produce New cell and cell components Chemicals Waste products Metabolites (products) A substance produced during cell processes

14 Primary and secondary metabolites
Primary metabolite Substance produced by organism as part of it’s normal growth E.g. amino acids, proteins, enzymes Production of primary metabolites matches the growth in population Secondary metabolite A substance only produced at a particular growth phase No direct involvement in fundamental metabolite processes Production usually begins after the main growth phase of the micro organisms

15 Batch culture Starter population is mixed with a specific quantity of nutrient solution Allowed to grow for a fixed period Products removed Fermentation tank emptied Examples Penicillin production Enzyme production

16 Continuous Culture Nutrients are added and products are removed from the fermentation tank at regular intervals Examples Insulin production from genetically modified E.Coli Production of mycoprotein

17 Learning outcomes Explain the importance of asepsis in the manipulation of microorganisms

18 Asepsis Asepsis Aseptic techniques absence of unwanted microorganisms
Any measure taken during a biotechnological process to prevent contamination by unwanted microorganisms

19 The importance of asepsis
Unwanted microorganisms Compete with the culture microorganisms Reduce the yield of useful products Cause spoilage of the product Produce toxic chemicals Destroy the culture microorganism or its products.

20 Methods to maintain asepsis
Ensure all fermenters and attachments are sterile Cleaning with pasteurised steam Chemical sterilisation Sterilise all liquids, solids and gases that enter the reaction vessel Maintain a pressure difference between the air in the room where fermentation is taking place and outside Maintains a steady airflow out of the room Ensure culture of microorganisms is pure Ensure the workers do not introduce unwanted microorganisms from their skin.

21 Learning Outcomes Describe how enzymes can be immobilised.
Explain why immobilised enzymes are used in large-scale production.

22 Immobilising enzymes Enzymes act as catalysts in metabolic reactions
Enzymes are useful in industrial processes Specificity Temperature of enzyme action Enzymes in solution need to be separated from the products. Immobilised enzymes can be re-used many times and leaves the product enzyme free.

23 Methods for immobilising enzymes
Gel entrapment Example – immobilising lactase in alginate Stages Enzyme solution is mixed with sodium alginate solution Droplets of this solution are added to a solution of calcium chloride The droplet turns into a bead which contains the enzyme

24 Immobilising lactase in alginate

25 Immobilising lactase in alginate
The beads can be tightly packed into a column The liquid substrate can be trickled over the beads The product trickles out of the bottom of the column The product is collected and purified.

26 Methods of immobilising enzymes
Adsorption / carrier bound Enzyme molecules are mixed with immobilising support e.g. glass beads or clay Covalent Bonding / cross-linked Enzyme molecules covalently bonded to a support

27 Methods of immobilising enzymes
Entrapment / inclusion Enzymes trapped in their natural state in a gel bead Reaction rate can be reduced as substrate needs to get through the trapping barrier Membrane separation Substrate separated from the mixture by a partially permeable membrane.

28 Advantages of immobilised enzymes
The advantages of using immobilised enzymes over enzymes in solution are Immobilised enzymes can be reused Product is enzyme free Immobilised enzymes are more tolerant to pH and temperature changes

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