Part 3: Genetic and Metabolic Engineering Flashcards
Def: Genetic Engineering
A set of technologies that directly manipulate and organisms genes, including transferring genes within and across species boundaries.
Def: Metabolic Engineering
Directed improvement of cellular properties through the modification of specific biochemical reactions, or the introduction of new ones, using recombinant DNA technology
Why are genetic and metabolic engineering approaches used
1) Make new/ more product
2) Improve product properties
3) Reduce waste
4) Reduce costs
Describe the key steps involved in creating recombinant insulin
1) Identify the insulin gene
2) PCR (genetic photocopying) isolates gene.
3) Restriction enzyme cuts open plasmid at specific site and the insulin gene is inserted. Plasmid is ‘zipped’ back up with ligase
4) Introduce the recombinant plasmid to E.coli through a heat shock
5) Culture E.coli cells in a bioreactor
6) Extraction and purification of insulin
How to check the success of genetic modification?
Compare molecule size
PCR
Antibiotics
Colour selection: No colour change, interrupted gene sequence or no plasmid
Def: Metabolism
The summation of all chemical reactions in an organism. It includes anabolic (energy using) and catabolism (energy producing)
Def: Intermediary metabolite
The product of one biochemical reaction that is the substrate for another biochemical reaction
Def: Fermentation
The growth of a large number of cells in a bioreactor
Describe lactic acid fermentation
Glucose - Pyruvate- Lactate
Describe Ethanol fermentation
Glucose - Pyruvate - Acetaldehyde - Ethanol
Describe the main steps of aerobic respiration
1) Glycolysis: Glucose to pyruvate
2) Link reaction: Links glycolysis with aerobic processes in the mitochondria, creation of acetyl CoA
3) Krebs Cycle: Acetyl CoA donates its acetyl group to oxaloacetate, forming citrate. Subsequent reduction reactions cause the production of hydrogen carriers.
4) Oxidative Phosphorylation: The movement of released electrons causes an electrochemical gradient, stimulating chemiosmosis and the production of ATP.
Describe the key concepts involved in the metabolic engineering of cells.
Overexpression of enzymes
Using enhanced enzymes
Creating a new metabolic pathway
Deleting branched metabolic pathways
Def: Metabolic pathway
Any sequence of feasible and observable biochemical reactions connecting specified input and output metabolites
Def: Pathway flux
The rate at which input metabolites are processed to output metabolites
Def: Rate determining step
The biochemical reaction with the slowest metabolic flux, and the reaction that is improved first.
