Luciana Flashcards
What is the role of metabolic engineering in industrial production? What are the
production factors that need to be optimized by metabolic engineering?
Yield, efficiency/productivity, and titer
The role of metabolic engineering in industrial production is to optimize the production of a product by optimizing the different factors that affect the production (pathways) and make it as economical as possible. It includes the design of, production of, and optimization of pathways. One main difference from systems biology is that the pathways are opimized.
Titer (i.e., production concentration), the rate (productiviry) and yield (product in mass/moles) have to be optimized.
Briefly explain how to determine metabolic fluxes in the central metabolism of
an organism.
Metabolic fluxes in central metabolism can be determined by using 13C in the carbon source of the organism, and following the trail using NMR, for example.
Compare the main differences, advantages, and disadvantages between
metabolic flux analysis and flux balance analysis.
Metabolic flux analysis (MFA) is a method for analyzing networks, where fluxes are determined experimentaly under different conditions to identify limiting steps. Flux balance analysis (FBA) uses computers, mathematics and algorithms for estimating fluzes under different conditions, and what happens if one aspect is adjusted, etc. FBA uses the data measured in MFA, and is quicker, however it is only predictions. MFA measures what actually happens.
Explain how novel metabolic pathways can be elucidated, and how production
of a certain metabolite can be improved in unknown/not characterized bacteria.
Using general reaction rules, and taking into account the structure and chemical properties of the substrates, enzymes and products can be used to elucidate new pathways, especially in combination with algorithms and other methematical/computational tools. In unknown organisms, random mutations can be generated by introducing mutagens (UV-radiation, toxins) to the bacteria to see if any of the mutations offer improvement of the desired function, in which case these are cultivated and used further.
Given the advantage of the available tools and background knowledge on the
metabolism of model organisms, e.g. Corynebaterium glutamicum, explain the
reasoning for exploring alternative and rather unestablished hosts in metabolic
engineering.
Other bacteria can still be better, like with regard to thermophilia, extremophiles, substrate usage, etc. Things that withstand extreme conditions are easier to have uncontaminated, and thus undisturbed.
. Briefly describe the steps that can be taken in systems metabolic engineering
towards large-scale, high yield production of value-added products from
microbial cells.
Pathway production/design - use reaction rules and chemical structure and properties - enzyme promiscuity, computers.
Pathway construction - can genetically modify the host organism, could utilize codon bias.
Pathway optimalization - increase host tolerance, codon bias, adaptive laboratory evolution, metabolic flux optimization, scaling up to industrial levels
