Team:TP CC-SanDiego/Project

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iGEM San Diego - Project

Background
The Project Background and Explanation

AflatoxinB1-Detoxifizyme is produced by Armillariella Tabescens intracellularly. Its gene is analyzed to have no introns and has been recombined into a plasmid before. Extraction is rather difficult and commercialization has not been able to be done yet.

Zearalenone Hydorlase is produced by Clonostachys Rosea. Its gene is also analyzed to have no introns and has been recombined into a plasmid before. Similar to ADTZ, commercialization has not been done yet.


Natural E. coli secretes various proteins by signal peptides that recognize translocons. We focus on the SEC pathway that lets the protein fold in the periplasm first, as it usually results in more correct folding, correct formation of disulfide bridges, minimizes degradation, and is less likely to result in accumulation.
Idea
E. Coli Capable of Extracelluar Secretion of Mycotoxin Detoxifying Enzymes
Our idea is to attach signal peptides to the N-terminus of the detoxifying enzymes. We use alpha amylase signal peptides and beta-lactamase signal peptides. The original proteins for these signal peptides are secreted out naturally by E. Coli, especially in strain K that we use for our project. They both follow sec-dependent pathways. In the case of beta-lactamase signal protein, cases have been shown of successful fusion proteins secreted by the beta-lactamase signal protein (link to: http://www.ncbi.nlm.nih.gov/pubmed/6607473). Successful continual excretion of the mycotoxin degradation enzymes can be purified in mass amounts periodically, or the chimeric E. Coli can be placed directly on crops to prevent
How
The Process to Complete our Project
Project 1 - 9