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 (PHAMCL) 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-3HAMCL) 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 3HAMCL using an unrelated carbon source, for the first time in a plasmid- and antibiotic-free bioprocess.

中文翻译：

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重组假单胞菌在无质粒和无抗生素培养物中生产聚羟基链烷酸酯共聚物。

将三种不同的多羟基链烷酸（PHA）合酶基因（富营养小球藻（Ralstonia eutropha H16），气单胞菌属TSM81或嗜水气单胞菌ATCC7966 phaC）引入到两个假单胞菌菌株的染色体中：天然中链长度的3-聚羟基链烷酸酯（PHAMCL）生产者（假单胞菌sp。 （LFM046）和UV诱导的突变株不能产生PHA（假单胞菌属sp。LFM461）。我们首次报道使用mini-Tn7转座子系统插入phaC的染色体副本。获得了稳定的无抗生素标记物和无质粒的重组体。随后，这些重组体使用葡萄糖作为唯一的碳源，通过这些重组体生产了P（3HB-co-3HAMCL），而无需共底物且在无抗生素的条件下。携带亲水链球菌phaC的重组体产生了由84.2 mol％的3-羟基丁酸酯组成的三元聚酯，仅来自葡萄糖的6.3摩尔％的3-羟基己酸酯和9.5摩尔％的3-羟基癸酸酯。因此，我们成功地增加了假单胞菌的工业潜力。通过使用不相关的碳源生产含3HB和3HAMCL的PHA共聚物，这是LFM461菌株，这是首次在无质粒和无抗生素的生物工艺中生产。