Team:Tec-Monterrey EKAM/Description

Abstract
The idea that a basic biological system could be used in the production of a related group of molecules suggests that this kind of manufacturing could be designed to optimally synthesize specific products from a wide range at a given time. The proper strategies for the modular design and execution of such projects are hereby addressed, making it possible to manipulate such modules in a project’s genetic construct with ease and resource frugality. A software tool is implemented for the appropriate design of a master DNA sequence with modular restriction sites placed in such a way that further cloning is facilitated. As proof of concept, a biological platform in the yeast Pichia pastoris is proposed for the production of terpenoids, useful hydrocarbons linked by the mevalonate pathway as their common source, focusing on the synthesis of lycopene. The restoration of an RNAi mechanism in the yeast by insertion of Saccharomyces castellii genes is also proposed for its use in silencing the ERG9 gene, responsible for the synthesis of the terpenoid squalene, further optimizing the platform. Additionally, the insertion of an N-terminus truncated enzyme from the mevalonate pathway to limit reported metabolic regulation is set forth, together with the use of the peroxisome targeting signal type 1 in the enzymes used. The need for reliable promoters for P. pastoris is addressed by characterizing three inducible promoters with GFP as a reporter.