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首页> 外文期刊>Journal of bacteriology >The Ralstonia eutropha PhaR Protein Couples Synthesis of the PhaP Phasin to the Presence of Polyhydroxybutyrate in Cells and Promotes Polyhydroxybutyrate Production
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The Ralstonia eutropha PhaR Protein Couples Synthesis of the PhaP Phasin to the Presence of Polyhydroxybutyrate in Cells and Promotes Polyhydroxybutyrate Production

机译：Ralstonia eutropha PhaR蛋白将PhaP蛋白的合成与细胞中多羟基丁酸酯的存在耦合，并促进多羟基丁酸酯的生产

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Polyhydroxyalkanoates (PHAs) are polyoxoesters that are produced by many bacteria and that accumulate as intracellular granules. Phasins (PhaP) are proteins that accumulate during PHA synthesis, bind PHA granules, and promote further PHA synthesis. Interestingly, PhaP accumulation seems to be strictly dependent on PHA synthesis, which is catalyzed by the PhaC PHA synthase. Here we have tested the effect of the Ralstonia eutropha PhaR protein on the regulation of PhaP accumulation. R. eutropha strains with phaR, phaC, and/or phaP deletions were constructed, and PhaP accumulation was measured by immunoblotting. The wild-type strain accumulated PhaP in a manner dependent on PHA production, and the phaC deletion strain accumulated no PhaP, as expected. In contrast, both the phaR and the phaR phaC deletion strains accumulated PhaP to higher levels than did the wild type. This result implies that PhaR is a negative regulator of PhaP accumulation and that PhaR specifically prevents PhaP from accumulating in cells that are not producing PHA. Transfer of the R. eutropha phaR, phaP, and PHA biosynthesis (phaCAB) genes into a heterologous system, Escherichia coli, was sufficient to reconstitute the PhaR/PhaP regulatory system, implying that PhaR both regulates PhaP accumulation and responds to PHA directly. Deletion of phaR caused a decrease in PHA yields, and a phaR phaP deletion strain exhibited a more severe PHA defect than a phaP deletion strain, implying that PhaR promotes PHA production and does this at least partially through a PhaP-independent pathway. Models for regulatory roles of PhaR in regulating PhaP and promoting PHA production are presented.
机译：聚羟基链烷酸酯（PHA）是由许多细菌产生并累积为细胞内颗粒的聚氧酸酯。钙蛋白（PhaP）是在PHA合成过程中积累，结合PHA颗粒并促进PHA进一步合成的蛋白质。有趣的是，PhaP积累似乎严格依赖于PhaC PHA合酶催化的PHA合成。在这里，我们测试了 Ralstonia eutropha PhaR蛋白对PhaP积累的调节作用。 R。构建了带有 phaR ， phaC 和/或 phaP 缺失的富营养菌株，并通过免疫印迹法检测了PhaP的积累。如预期的那样，野生型菌株以依赖PHA产生的方式积累PhaP，而 phaC 缺失菌株没有积累PhaP。相反， phaR 和 phaR phaC 缺失菌株的PhaP积累水平均高于野生型。该结果暗示PhaR是PhaP积累的负调节剂，并且PhaR特异性地阻止了PhaP在不产生PHA的细胞中积累。 R的转移。 eutropha phaR ， phaP 和PHA生物合成（ phaCAB ）基因进入异源系统大肠杆菌，足以重组PhaR / PhaP调节系统，意味着PhaR既调节PhaP积累又直接响应PHA。删除 phaR 导致PHA产量下降，并且 phaR phaP 缺失菌株表现出比 phaP 缺失菌株更严重的PHA缺陷，这表明PhaR促进了PHA的产生，并至少部分地通过非PhaP依赖性途径进行。提出了PhaR在调节PhaP和促进PHA产生中的调节作用模型。

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