Full text
| |
| Author(s): |
Edmar Ramos de Oliveira Filho
Total Authors: 1
|
| Document type: | Master's Dissertation |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) |
| Defense date: | 2017-02-02 |
| Examining board members: |
Luiziana Ferreira da Silva;
Maria Beatriz Riveron Acosta;
Regina Lúcia Baldini;
Patrícia Locosque Ramos
|
| Advisor: | Luiziana Ferreira da Silva |
| Abstract | |
Polyhydroxyalkanoates (PHA) are biopolymers naturally produced and accumulated by many organisms such as bacteria, archaeas and some eukaryotes, such as fungi and yeasts. As thermoplastics, biodegradable, biocompatible and possibly made from renewable resources, they exhibit great potential to replace oil-derived plastics. Hybrid PHA copolymers can be formed by short and medium-chain monomers, P(3HASCL-co-3HAMCL), and present industry desired physicochemical properties, becoming similar to conventional oil-based plastics. PHA synthase is the key enzyme in PHA biosynthesis, responsible for catalyzing the polymerization of (R)-hydroxyacyl-CoA molecules, influencing polymer monomeric composition. Tn7-based recombination strategies represent powerful molecular tolls designed for gene delivery in Gram-negative bacteria, as mini-Tn7 elements can be readily transferred to recombinants production. In this work, its presented the constructions of recombinant Pseudomonas strains harboring PHA synthase genes from Ralstonia eutropha and Aeromonas strains in specific sites of its chromosome, and the production of P(3HB-co-3HO-co-3HD). Obtained clones were screened to confirm chromosomic insertion of the phaC sequences. Positive clones PHA production and composition were evaluated in shaken-flasks assays using glucose as the only carbon source. One of the constructed recombinants accumulated P(3HB-co-3HO-co-3HD), corresponding about to 2 % of its Cell Dry Weight as PHA. (AU) | |
| FAPESP's process: | 14/15448-0 - Construction of recombinant pseudomonas strains harboring different PHA synthases in its genome to produce 3HB-co-3HAMCL |
| Grantee: | Edmar Ramos de Oliveira Filho |
| Support Opportunities: | Scholarships in Brazil - Master |