Production of Polyhydroxyalkanoates Copolymers by Recombinant Pseudomonas in Plasmid- and Antibiotic-Free Cultures

Author(s)

Oliveira-Filho, Edmar Ramos ^[1] ; Guaman, Linda P. ^{[1, 2]} ; Mendonca, Thatiane Teixeira ^[1] ; Long, Paul F. ^{[3, 4]} ; Taciro, Marilda Keico ^[1] ; Cabrera Gomez, Jose Gregorio ^[1] ; Silva, Luiziana F. ^[1]

Affiliation

^[1] Univ Sao Paulo, Inst Biomed Sci, Cidade Univ, Sao Paulo - Brazil

^[2] Univ UTE, Ctr Invest Biomed, Fac Ciencias Salud Eugenio Espejo, Quito - Ecuador

^[3] Kings Coll London, Sch Canc & Pharmaceut Sci, London - England

^[4] Univ Sao Paulo, Sch Pharmaceut Sci, Cidade Univ, Sao Paulo - Brazil

• Document type: Journal article
• Source: JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY; v. 28, n. 5, p. 225-235, 2018.
• Web of Science Citations: 0

Abstract

Three different polyhydroxyalkanoate (PHA) synthase genes (Ralstonia eutropha H16, Aeromonas sp. TSM81 or Aeromonas hydrophila ATCC7966 phaC) were introduced into the chromosome of two Pseudomonas strains: a native medium-chain-length 3-polyhydroxyalkanoate (PHA(MCL)) producer (Pseudomonas sp. LFM046) and a UV-induced mutant strain unable to produce PHA (Pseudomonas sp. LFM461). We reported for the first time the insertion of a chromosomal copy of phaC using the transposon system mini-Tn7. Stable antibiotic marker-free and plasmid-free recombinants were obtained. Subsequently, P(3HB-co-3HA(MCL)) was produced by these recombinants using glucose as the sole carbon source, without the need for co-substrates and under antibiotic-free conditions. A recombinant harboring A. hydrophila phaC produced a terpolyester composed of 84.2 mol% of 3-hydroxybutyrate, 6.3 mol% of 3-hydroxyhexanoate, and 9.5 mol% of 3-hydroxydecanoate from only glucose. Hence, we were successful in increasing the industrial potential of Pseudomonas sp. LFM461 strain by producing PHA copolymers containing 3HB and 3HA(MCL) using an unrelated carbon source, for the first time in a plasmid-and antibiotic-free bioprocess. (c) 2019 S. Karger AG, Basel (AU)

• FAPESP's process: 12/51533-6 - Tailor-made biodegradable polymers produced ·by bacteria from biomass
• Grantee: Luiziana Ferreira da Silva
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Regular Program Grants
• FAPESP's process: 10/51692-1 - Metabolic engineering applied to biopolymer production
• Grantee: Marilda Keico Taciro
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Young Investigators Grants
• 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