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 Opportunities: Program for Research on Bioenergy (BIOEN) - Regular Program Grants
• Start date: August 01, 2016
• End date: July 31, 2019
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Luiziana Ferreira da Silva
• Grantee: Luiziana Ferreira da Silva
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: José Gregório Cabrera Gomez ; Maria Manuela Regalo da Fonseca ; Maria Teresa Ferreira Cesário Smolders ; Marilda Keico Taciro ; Rubén Jesus Sánchez Rodríguez

Abstract

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)

Scientific publications

(5)

(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, MAR 2018. (10/51989-4, 16/00842-0)

OLIVEIRA-FILHO, EDMAR R.; DE MACEDO, MATHEUS A.; LEMOS, ALINE C. C.; ADAMS, FRIEDERIKE; MERKEL, OLIVIA M.; TACIRO, MARILDA K.; GOMEZ, JOSE GREGORIO C.; SILVA, LUIZIANA F.. Engineering Burkholderia sacchari to enhance poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] production from xylose and hexanoate. International Journal of Biological Macromolecules, v. 213, p. 13-pg., 2022-06-13. (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, OCT 2021. (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, MAY 15 2018. (10/51989-4, 16/00842-0)

OLIVEIRA-FILHO, EDMAR R.; SILVA, JEFFERSON G. P.; DE MACEDO, MATHEUS ARJONA; TACIRO, MARILDA K.; GOMEZ, JOSE GREGORIO C.; SILVA, LUIZIANA F.. Investigating Nutrient Limitation Role on Improvement of Growth and Poly(3-Hydroxybutyrate) Accumulation by Burkholderia sacchari LMG 19450 From Xylose as the Sole Carbon Source. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, v. 7, p. 11-pg., 2020-01-08. (16/00842-0)

Filed patent(s) as a result of this research project