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Identification and Isolation of Genes Involved in Poly(β-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

机译:巴西固氮螺菌中聚(β-羟基丁酸)生物合成相关基因的鉴定与分离及phbC突变体的表征

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摘要

Like many other prokaryotes, rhizobacteria of the genus Azospirillum produce high levels of poly(β-hydroxybutyrate) (PHB) under suboptimal growth conditions. Utilization of PHB by bacteria under stress has been proposed as a mechanism that favors their compatible establishment in competitive environments, thus showing great potential for the improvement of bacterial inoculants for plants and soils. The three genes that are considered to be essential in the PHB biosynthetic pathway, phbA (β-ketothiolase), phbB (acetoacetyl coenzyme A reductase), and phbC (PHB synthase), were identified in Azospirillum brasilense strain Sp7, cloned, and sequenced. The phbA, -B, and -C genes were found to be linked together and located on the chromosome. An A. brasilense phbC mutant was obtained by insertion of a kanamycin resistance cassette within the phbC gene. No PHB production was detected in this mutant. The capability of the wild-type strain to endure starvation conditions was higher than that of the mutant strain. However, motility, cell aggregation, root adhesion, and exopolysaccharide (EPS) and capsular polysaccharide (CPS) production were higher in the phbC mutant strain than in the wild type.
机译:像许多其他原核生物一样,Azospirillum属的根瘤菌在次佳生长条件下也能产生高水平的聚(β-羟基丁酸酯)(PHB)。已经提出了在压力下细菌利用PHB作为有利于它们在竞争环境中相容建立的机制,因此显示出改善植物和土壤细菌接种剂的巨大潜力。在巴西假单胞菌属菌株Sp7中鉴定了被认为是PHB生物合成途径必不可少的三个基因phbA(β-酮硫解酶),phbB(乙酰乙酰辅酶A还原酶)和phbC(PHB合酶),并对其进行了克隆和测序。发现phbA,-B和-C基因连接在一起并位于染色体上。通过在phbC基因内插入卡那霉素抗性盒获得巴西农杆菌phbC突变体。在该突变体中未检测到PHB产生。野生型菌株忍受饥饿条件的能力高于突变菌株。但是,phbC突变菌株的运动性,细胞聚集,根系粘附以及胞外多糖(EPS)和荚膜多糖(CPS)的产量高于野生型。

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