Certificate One Marketing Introduction to Metabolic Engineering Summary
The key points from this module are:
Metabolic Engineering is a process for modification of specific biochemical reactions of microorganisms to produce the required amounts of the desired metabolites via recombinant DNA technology.
Steps in metabolic engineering:
1. Host selection
2. Selection of the pathway
3. Parts assembly
4. Testing via spectroscopic analysis
The interrelationship between genetic engineering and metabolic engineering:
Genetic Engineering allowed precise modification of specific enzymatic reactions through specific genetic perturbations which helps to modify the promoter strength of a given gene. It is used for gene deletions and introduction of new genes or pathways into the cells.
Metabolic Engineering employ directed improvement of products formation or cellular properties via modification of specific biochemical reactions, or introduction of new heterozygous gene expression via the use of recombinant DNA technology.
Metabolic Engineering involves mechanisms for effecting over expression of regulator, eliminating bottlenecks, improving enzyme specificity and blocking competing pathway.
Genetic Engineering helps in heterologous products synthesis, mutagenesis and selection, and genome sequencing. It also helps in protein engineering and laboratory evolution to deliver enzymes which aid production of new products.
The Metabolic Engineering Cycle - The pictorial representation of the metabolic cycle process is as shown:
The major advantages of E. coli as a microbial cell factory:
1. High growth rate.
2. Availability of gene and genome engineering tools.
3. Established high cell culture techniques.
4. Various metabolic engineering tools e.g. the genome-scale metabolic models (GEMs).
5. Established high cell density culture for E. coli.
The basic elements of metabolic engineering:
Integrated metabolic pathways deals with:
Pathway flux and control
Understanding complex enzyme networks requires:
Enzymology of participating enzymes.
Knowledge of the enzymatic pathways.
Pathway chemistry and stoichiometry.
Computer-aided biochemical pathways requires:
The expected maximal yields.
Methods of bypassing critical bottlenecks.
Identified key intermediates.
Metabolic flux analysis - involves the determination of intracellular fluxes, and analysis of factors affecting flux distributions.
Metabolic flux analysis provides data about the following:
Uptake and secretion rates.
4. Quasi-steady-state mass balance on metabolic intermediates to determine intercellular metabolic fluxes.
The information obtained from flux analysis can be used for:
Identifying critical branch points in the pathway.
Discover unusual pathways n less characterized species.
3. Define the maximum theoretical yield for the synthesis of products from complex integrated pathways.
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