A non-naturally-occurring P(3HB-co-3HAMCL) is produced by recombinant Pseudomonas sp. from an unrelated carbon source

Cespedes, Lucas G.; Nahat, Rafael A.T.P.S.; Mendonça, Thatiane Teixeira; Tavares, Rafaela R.; Oliveira-Filho, Edmar R.; Silva, Luiziana Ferreira da; Taciro, Marilda Keico; Sánchez, R.; Gomez, José Gregório Cabrera

doi:10.1016/j.ijbiomac.2018.03.051

Cited by 12 publications

(8 citation statements)

References 47 publications

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“…Despite the similar HA MCL content accumulated by our stable recombinant strain (15.8 mol%), we obtained not only 3HHx, but also 3HD when glucose was supplied as the sole carbon source (Table 1), without the need of co-substrates. This extends the results previously presented by Cespedes et al [2018] in which differential scanning calorimetry analysis evidenced the successful incorporation of 3HB monomers. Table 2 summarizes the production of PHA from various carbon sources by using different bacteria reported in the literature.…”

Section: Resultssupporting

confidence: 91%

“…We have overcome this impediment by demonstrating for the first time the successful expression of a phaC gene when integrated at the attTn7 site of the Pseudomonas chromosome, and then growing the recombinant strain on glucose, in a plasmid-free bioprocess without the addition of antibiotics. This research extends the results previously reported by Cespedes et al [2018].…”

Section: Discussionsupporting

confidence: 93%

“…3) chromosome. A similar result was recently obtained by Cespedes et al [2018], which used Pseudomonas sp. LFM461 as the host for a medium-copy number plasmid pBBR1MCS-2 harboring phaPCJ TSM81 genes and the same carbon substrate (glucose).…”

Section: Resultssupporting

confidence: 86%

“…Additionally, our results reinforce that Pseudomonas spp. LFM046 and LFM461 native metabolic networks supply 3HB (HA SCL ) and different 3HA MCL monomers, such as 3HHx and 3HD, recently evidenced by Cespedes et al [2018].…”

Section: Resultsmentioning

confidence: 97%

“…LFM461, a PHA-negative mutant of Pseudomonas sp. LFM046 [Gomes, 2009;Cespedes et al, 2018]. This heterologous complementation indicates the mutation preventing it from producing PHA is located at the gene encoding its native PHA synthase.…”

Section: Introductionmentioning

confidence: 89%

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

et al. 2018

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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.

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Section: Resultssupporting

confidence: 91%

Section: Discussionsupporting

confidence: 93%

Section: Resultssupporting

confidence: 86%

Section: Resultsmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 89%

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

et al. 2018

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Cross‐sectoral citrus sugarcane biorefinery: polyhydroxyalkanoates production in Brazil

Kanbe,

da Silva,

Gomez

2023

Biofuels Bioprod Bioref

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Polyhydroxyalkanoates (PHA) are naturally occurring polyesters accumulated by bacteria under conditions of inorganic nutrient limitations and excess carbon. They offer renewable and biodegradable alternatives to petroleum‐based polymers, especially those used in transient single‐use applications. However, PHA still faces high production costs, primarily linked to the carbon source, lower productivity in processes and an early stage of technical development in comparison with well‐established fossil‐based polymers, which challenge its transition to a true commodity market. Recent research has highlighted the potential of renewable, inexpensive, abundant and non‐edible carbon‐rich residues from agriculture, municipal solid waste and industrial byproducts as alternative carbon sources in developing the production process. Recognizing Brazil's position as the leading producer of both citrus and sugarcane, this review introduces a novel citrus–sugarcane biorefinery model for PHA production, capitalizing on the geographical and operational synergies of both sectors. The proposed model integrates wet citrus pulp as a second‐generation carbon source, sugarcane industry fermenters, waste management, solvents and renewable energy from both sectors. Primary challenges involve the need for efficient, low‐cost biomass pre‐treatment to remove limonene and partially extract citric pectin, coupled with the partial hydrolysis of pectin to enhance solubility and reduce viscosity. Additional considerations include determining citrus biomass composition and sugar consumption, screening bacterial diversity for optimal PHA production, and deepening our understanding of the metabolic pathways underpinning the catabolism of citrus residues. Future research should focus on enhancing PHA concentrations and diversity and finally conducting life cycle assessment studies to evaluate the sustainability and feasibility of this proposed model.

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Metabolic engineering for biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from glucose and propionic acid in recombinant Escherichia coli

Meng

Miao

Liu

et al. 2022

Bioresource Technology

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A non-naturally-occurring P(3HB-co-3HAMCL) is produced by recombinant Pseudomonas sp. from an unrelated carbon source

Cited by 12 publications

References 47 publications

Production of Polyhydroxyalkanoates Copolymers by Recombinant <b><i>Pseudomonas</i></b> in Plasmid- and Antibiotic-Free Cultures

Production of Polyhydroxyalkanoates Copolymers by Recombinant <b><i>Pseudomonas</i></b> in Plasmid- and Antibiotic-Free Cultures

Cross‐sectoral citrus sugarcane biorefinery: polyhydroxyalkanoates production in Brazil

Metabolic engineering for biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from glucose and propionic acid in recombinant Escherichia coli

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