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| Author(s): |
Rafael Augusto Theodoro Pereira de Souza Nahat
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) |
| Defense date: | 2019-07-05 |
| Examining board members: |
Marilda Keico Taciro;
Elisabeth de Fatima Pires Augusto;
Joao Meidanis;
Paulo Vieira Milreu;
Pedro de Alcântara Pessoa Filho;
Aldo Tonso
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| Advisor: | Marilda Keico Taciro; José Gregorio Cabrera Gomez |
| Abstract | |
A large database of chemostat cultures of the strain Pseudomonas sp. LFM046, an efficient polyhydroxyalkanoate (PHA) producer, was available and used to validate predictive genomescale metabolic models. Aiming to maximize the PHA content while controlling its monomeric composition (and hence its mechanical properties), three steps were taken. First, an N-phenotypic model was developed based on three biological principles and then applied to select the cultivations which best represent all possible phenotypes of the strain LFM046. Second, a draft genome-scale reconstruction of LFM046 generated automatically by the webservice KBase from the genome, assembled by the software MeDuSa, was refined using two methods: the usual manual method, systematized and logged for future reference, and NGlobalFit, a novel automated method which was validated as a proof-of-concept. The predictive qualities of the refined metabolic networks were assessed using the N-phenotypic model. Third, specific FBA (Flux Balance Analysis) simulations in these networks confirmed that the relative speed of the fatty-acids de novo synthesis is affected by culture conditions and determines the PHA content and composition. The methods and software developed in this work, in collaboration with the French research group European Research Team in Algorithms and Biology, formaL and Experimental (ERABLE), were generalized from the case-study of PHA production by Pseudomonas sp. LFM046 in order to stablish an R&D framework transferable to other strains and bioprocesses. (AU) | |
| FAPESP's process: | 16/01756-0 - Metabolic modelling of polihydroxyalkanoates production production with different monomeric compositions by Pseudomonas sp. |
| Grantee: | Rafael Augusto Theodoro Pereira de Souza Nahat |
| Support Opportunities: | Scholarships in Brazil - Doctorate (Direct) |