Team:PrepaTec GarzaSadaMx/Summary

Summary

The purpose of this experiment is to obtain Tyrian purple in a more environmentally friendly and efficient way than the one commonly used. The mollusk Bolinus Brandaris is killed in order to obtain this dye, thus we used the recombinant DNA techniques in order for the E. coli to produce Tyrian purple. We inserted two genes in E. coli for it to generate two enzymes which induce the reactions that produce indigo. After a chemical process, known as halogenation of benzene, indigo is turned into Tyrian purple. This process needs the usage of a catalyst called FeB3 in order for it to occur. Because we believe it´s essential that these species are kept from being endangered. The importance of this is related to the fact that it´s significant to maintain the biodiversity, as a way of preventing the disruption of the balance of the ecosystems.

This is a summary of the reactions occurring during the production of Tyrian purple:

Trypthophan (tryptophanase)-> indol (naphthalene dioxygenase) -> 2,3 dihidroxindolina -> indigo -> tyrian purple

The trypthophan and tryptophanase are already in E. coli, so the next step is to insert a plasmid in the E. coli, which contains the gen to produce the enzyme naftaleno diogenasa. To achieve this first we have to make competent cells, then transform the E. coli, and after that, we can recover the plasmid trough electrophoresis or miniprep in order to obtain the DNA we want. When we already have the enzyme naftaleno diogenasa, it turns the indole into 2,3 dihidroxindolina and in an spontaneous reaction with oxygen, we obtain indigo. After obtaining indigo we use a process called halogenation of benzene in order to get to the Tyrian purple.

Unfortunately, we tried but couldn't obtain the genes necessary for the production of the dye. Synthesis was out of our budget, so we decided to carry out the same procedural steps only with parts we already had from the iGEM Kit. We prepared chemocompetent cells and transformed three different E. coli with genes for Red Fluorescent Protein, Wintergreen odor generator and Banana odor generator. We performed a plasmid extraction (mini-prep) and visualized the ADN with gel electrophoresis.