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Team:Wageningen UR/Why Aspergillus nigem
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| - | <p> The production of these secondary metabolites in most cases involves a large backbone enzyme that contains multiple catalytic domains. One of our goals is to establish a modular system of domain shuffling to generate a plethora of novel enzymes with new and improved functionalities. The possibilities are endless as there are various different domains from fungi that can be added, removed, reordered or | + | <p> The production of these secondary metabolites in most cases involves a large backbone enzyme that contains multiple catalytic domains. One of our goals is to establish a modular system of domain shuffling to generate a plethora of novel enzymes with new and improved functionalities. The possibilities are endless as there are various different domains from fungi that can be added, removed, reordered, or even customized by this modularity approach.</p> |
| - | <p>For instance, one of the main enzymes involved in the production of lovastatin is the 277kDa LovB enzyme, which contains 7 different catalytic domains. Prior studies on this pathway provide a good insight into the catalytic mechanisms of these individual domains. The individual domains have been mapped by homology modeling and are found to be well defined as well, which makes the | + | <p>For instance, one of the main enzymes involved in the production of lovastatin is the 277kDa LovB enzyme, which contains 7 different catalytic domains. Prior studies on this pathway provide a good insight into the catalytic mechanisms of these individual domains. The individual domains have been mapped by homology modeling and are found to be well defined as well, which makes the modularity approach more feasible.</p> |
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Revision as of 17:50, 1 October 2013
- Why Aspergillus nigem?
- Secondary metabolites
- Toolbox
- Host engineering
- Summary
- Safety introduction
- General safety
- Fungi-related safety
- Biosafety Regulation
- Safety Improvement Suggestions
- Safety of the Application
- Why Aspergillus nigem?
- Secondary metabolites
- Lovastatin
- ATP Biosensor
- pH Biosensor
- Cytoskeleton and Septa
- Chromoproteins
- Host engineering
- Summary
Why Aspergillus nigem?
Cause there ain't no party like an Aspergillus party!
A Versatile and Powerful Synbio Host
Biotechnology has brought about a revolution in drug manufacturing since its inception. Many life-saving drugs have spun out of biotech companies over the past few decades. However, there is still a vast body of unexplored compounds such as the fungal secondary metabolites that have the potential to prolong human lifespan.
The production of lovastatin, a drug used in lowering LDL cholesterol for patients suffering from cardiovascular disease, has been chosen as the Herculean task for our team. Currently, this secondary metabolies-lovastatin is produced in fungi Aspergillus terreus, which also produces less desirable toxins. We aim to transfer the entire lovastatin metabolic pathway from A. terreus into a GRAS organism like Aspergillus niger. A. niger is amenable to genetic engineering which has already represented the most efficient, highest yielding bioprocess in practice for organic acids production.
Modular Structure of DNA Sequence
The production of these secondary metabolites in most cases involves a large backbone enzyme that contains multiple catalytic domains. One of our goals is to establish a modular system of domain shuffling to generate a plethora of novel enzymes with new and improved functionalities. The possibilities are endless as there are various different domains from fungi that can be added, removed, reordered, or even customized by this modularity approach.
For instance, one of the main enzymes involved in the production of lovastatin is the 277kDa LovB enzyme, which contains 7 different catalytic domains. Prior studies on this pathway provide a good insight into the catalytic mechanisms of these individual domains. The individual domains have been mapped by homology modeling and are found to be well defined as well, which makes the modularity approach more feasible.
A Toolkit for A. niger
In addition to the modularity approach, a toolkit for A. niger is also being created, consisting of an ATP and a pH biosensor that can be targeted to specific compartments with the use of signal peptides, actin and septa GFP fusions to visualize the cytoskeleton and the junctions between adjacent cells, and chromoproteins that can serve as secondary selection markers. These tools allow one to obtain insight in cellular economy, physiology and architecture, which can be used to optimize the production processes in A. niger.
