Team:Concordia/FutureWork

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  • Lactococcus Lactis
  • Scaffold
  • Food Intolerances
  • Prototype Pathway
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Future Work

As with any new technology, it is important to understand that the scaffold has certain limitations which must be improved upon.

For example, an increase in the number of cohesin products expressed in a scaffold displayed by modified L. lactis greatly reduces the total efficiency of display (1). This in turn, reduces the total amount of scaffolds displayed on a bacterium at a determined moment. One of the goals of future work would be to increase the efficiency of expression of the scaffold, so as to enable it to synthesize and produce larger platforms for protein expression. This could have a big impact on efficiency, since more enzymes displayed by an organism could mean an augmented yield of the product of interest. At the same time, with more cohesin products on the scaffold, larger and more intricate pathways could be developed and implemented without compromising efficiency or feasibility.

There exist also many interesting questions that once solved could help increase the utility and usefulness of the scaffold. For example, it would be very interesting to explore the sizes of enzymes that are to be harbored by the scaffold, to determine limits for their display. That is to say, to explore the maximum and minimum ranges at which it is advantageous to display a protein on the scaffold, and also the size of enzymes at which steric hindrance would become important enough to reduce the efficiency of the processes performed by the scaffold. This, of course, would be followed by further investigation to be able to extend these limits and display very large protein molecules that could be of interest for their catalytic abilities and that could benefit from being expressed extracellularly in the fashion proposed by this project.

In a similar way, it would be of great interest to explore different quaternary structures that could be displayed on the scaffold, along with their respective efficiencies of display and assembly. In this work, we limited ourselves to the use of monomeric proteins, since the use of other structural types of protein could be detrimental for the demonstration of the efficiency of the metabolic pathway proposed. It was unknown to us how the dockerin domain in the primary structure of the recombinant proteins would affect binding to other monomers and, hence, how this altered quaternary structure would affect the efficiency of our proposed metabolic pathway. Future endeavors would include the exploration and further analyses of these polymeric structures and their role in the efficiency and plausibility of application onto the scaffold.

Many different areas of industry can benefit by the use of the scaffold in synthetic processes. Increased efficiency of biofuel production from renewable sources could be potentially achieved by the utilizing scaffolds. This would mean, in principle, higher yields of production and the instauration of more efficient, and greener processes of mass transformation.

Other areas of science and technology can also take advantage of our work, like the health sector, which could use engineered, scaffold-displaying bacteria to act as a system of continuous drug delivery by attaching enzymes capable of synthesizing compounds from substrates readily available in the proximity of the bacterial colonies. Modified organisms could also act as probiotics to enhance a person’s capability to degrade certain compounds that would be otherwise harmful. For example, the use of a recombinant version of lactase that could be displayed on the scaffold for the treatment and prevention of lactose intolerance.

There exists a multitude of other interesting areas of research to be explored involving the use of scaffolds. attaching scaffolds onto other scaffolds to increase the number of spaces available for the expression of enzymes on a single organism. The use of scaffold-scaffold interactions to form matrices of organisms could be used in many different ways, for instance in tissue regeneration. Further engineering of scaffolds to attach themselves to harmful agents in the bloodstream of an organism would be another interesting path to follow, with the purpose of acting as antibodies for the enhancement of the action in the immune response. These are just a few of many possible beneficial applications of scaffolds, thus, we believe that pursuing research in this area could have a positive impact on the scientific community and be very rewarding to motivated future iGEM teams.

Reference

1	Badilak, SF et al., Reprint of: Extracellular matrix as a biological scaffold material: Structure and function