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Enhancing pentose and co-substrate bacterial uptake for the production of biopolymers from biomass: strain improvement, bioprocess optimization and application tests

Grant number: 16/00842-0
Support type:Program for Research on Bioenergy (BIOEN) - Regular Program Grants
Duration: August 01, 2016 - July 31, 2019
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal researcher:Luiziana Ferreira da Silva
Grantee:Luiziana Ferreira da Silva
Home Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Assoc. researchers:José Gregório Cabrera Gomez ; Maria Manuela Regalo da Fonseca ; Maria Teresa Ferreira Cesário Smolders ; Marilda Keico Taciro ; Rubén Jesus Sánchez Rodríguez


This proposal focuses the development of efficient bacterial strains to utilize the sugars from hemicelulosic hydrolysates and to produce the biodegradable and biocompatible polymers polyhydroxyalkanoates (PHA). The target PHA is PHB-co-HHx, a copolymer with 3-hydroxybutyrate (HB) and 3-hydroxyhexanoate (HHx), exhibiting sufficient flexibility for medical applications, becoming more flexible as the HHx content increases. Owing to its properties there are big companies investing in the production of this copolymer and some patents being deposited. Previous works indicated that recombinants of the highly PHA-accumulating strain B. sacchari, harboring appropriate genes from Aeromonas sp, were able to produce PHB-co-HHx, from sugars and hexanoate, with increased amounts of HHx (up to 15 times more than wild type). Since only 50% of hexanoate was transformed into HHx, further bacterial improvement is sought in this project, with the possibility to modulate the HHx content, enhancing the range of applications. We also previously demonstrated that in B. sacchari , despite being one of the Best xyloseutlizing bacteria, conversion of xylose to bioproducts can be improved and this aspect will also be explored in the recombinants to be constructed. Once an appropriate microorganism is obtained, fed-batch cultivations in bioreactor will be developed and the best strategy for both carbon-source and cosubstrate (hexanoate) addition will be sought to attain maximum productivities of PHB-co-HHx containing high HHX incorporations. Metabolic flux analysis will be applied in all steps to evaluate the micro-organisms studied, the influence of the genetic modifications introduced and also the bioprocess conduction, resulting in information to give a feedback to further modifications, if needed. Copolymers will be extracted and properties will be evaluated for medical applications. Our group from ICB USP is associated to IBB IST* (Instituto de Biotecnologia e Bioengenharia, Instituto Superior Técnico, Universidade de Lisboa, Portugal), LAMAV UENF** (Laboratorio de Materiais Avançados, Univ. Estadual Norte Fluminense, RJ, Brazil) and the Westminster University (UK) to develop bioreactor (& extraction) experiments, polymer characterization and medical applications, respectively. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
GUAMAN, LINDA P.; OLIVEIRA-FILHO, EDMAR R.; BARBA-OSTRIA, CARLOS; GOMEZ, JOSE G. C.; TACIRO, MARILDA K.; DA SILVA, LUIZIANA FERREIRA. xylA and xylB overexpression as a successful strategy for improving xylose utilization and poly-3-hydroxybutyrate production in Burkholderia sacchari. Journal of Industrial Microbiology & Biotechnology, v. 45, n. 3, p. 165-173, . (10/51989-4, 16/00842-0)
OLIVEIRA-FILHO, EDMAR RAMOS; GOMEZ, JOSE GREGORIO CABRERA; TACIRO, MARILDA KEICO; SILVA, LUIZIANA FERREIRA. Burkholderia sacchari (synonym Paraburkholderia sacchari): An industrial and versatile bacterial chassis for sustainable biosynthesis of polyhydroxyalkanoates and other bioproducts. Bioresource Technology, v. 337, . (16/00842-0)
GUAMAN, LINDA P.; BARBA-OSTRIA, CARLOS; ZHANG, FUZHONG; OLIVEIRA-FILHO, EDMAR R.; GOMEZ, JOSE GREGBRIO C.; SILVA, LUIZIANA F.. Engineering xylose metabolism for production of polyhydroxybutyrate in the non-model bacterium Burkholderia sacchari. Microbial Cell Factories, v. 17, . (10/51989-4, 16/00842-0)

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