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

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY; v. 28, n. 5, p. 225-235, 2018.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 10/51935-1 - Estudo de bacterias recombinantes e analise de fluxos metabolicos para a biossintese do copolimero biodegradavel poli(3-hidroxibutirato-co-3-hidroxihexanoato)[p(3hb-co-3hhx)]
• Beneficiário: Thatiane Teixeira Mendonça
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 12/51533-6 - Polímeros biodegradáveis feitos sob medida por bactérias a partir de biomassa
• Beneficiário: Luiziana Ferreira da Silva
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Regular
• Processo FAPESP: 10/51692-1 - Engenharia metabólica aplicada à produção de biopolímero
• Beneficiário: Marilda Keico Taciro
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Jovens Pesquisadores
• Processo FAPESP: 14/15448-0 - Construção de mutantes de pseudomonas abrigando diferentes PHA sintases em seu genoma, para produção de 3HB-co-3HAMCL
• Beneficiário: Edmar Ramos de Oliveira Filho
• Modalidade de apoio: Bolsas no Brasil - Mestrado