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首页> 外文期刊>Applied Microbiology and Biotechnology >Cloning, characterization and comparison of the Pseudomonas mendocina polyhydroxyalkanoate synthases PhaC1 and PhaC2
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Cloning, characterization and comparison of the Pseudomonas mendocina polyhydroxyalkanoate synthases PhaC1 and PhaC2

机译：Mendocina假单胞菌多羟基链烷酸合酶PhaC1和PhaC2的克隆，鉴定和比较

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摘要

This study describes a comparison of the polyhydroxyalkanoate (PHA) synthases PhaC1 and PhaC2 of Pseudomonas mendocina. The P. mendocina pha gene locus, encoding two PHA synthase genes [phaC1Pm and phaC2Pm flanking a PHA depolymerase gene (phaZ)], was cloned, and the nucleotide sequences of phaC1Pm (1,677 bp), phaZ (1,034 bp), and phaC2Pm (1,680 bp) were determined. The amino acid sequences deduced from phaC1Pm and phaC2Pm showed highest similarities to the corresponding PHA synthases from other pseudomonads sensu stricto. The two PHA synthase genes conferred PHA synthesis to the PHA-negative mutants P. putida GPp104 and Ralstonia eutropha PHB–4. In P. putida GPp104, phaC1Pm and phaC2Pm mediated PHA synthesis of medium-chain-length hydroxyalkanoates (C6–C12) as often reported for other pseudomonads. In contrast, in R. eutropha PHB–4, either PHA synthase gene also led to the incorporation of 3-hydroxybutyrate (3HB) into PHA. Recombinant strains of R. eutropha PHB–4 harboring either P. mendocina phaC gene even accumulated a homopolyester of 3HB during cultivation with gluconate, with poly(3HB) amounting to more than 80% of the cell dry matter if phaC2 was expressed. Interestingly, recombinant cells harboring the phaC1 synthase gene accumulated higher amounts of PHA when cultivated with fatty acids as sole carbon source, whereas recombinant cells harboring PhaC2 synthase accumulated higher amounts when gluconate was used as carbon source in storage experiments in either host. Furthermore, isogenic phaC1 and phaC2 knock-out mutants of P. mendocina provided evidence that PhaC1 is the major enzyme for PHA synthesis in P. mendocina, whereas PhaC2 contributes to the accumulation of PHA in this bacterium to only a minor extent, and then only when cultivated on gluconate.
机译：这项研究描述了假单胞菌假单胞菌的多羟基链烷酸酯（PHA）合成酶PhaC1和PhaC2的比较。克隆了编码两个PHA合酶基因[PHA解聚酶基因（phaZ）两侧的phaC1Pm 和phaC2Pm ]的P. mendocina pha基因位点，并确定了phaC1Pm 的核苷酸序列（1,677确定了bpZ，1,034 bp和phaC2Pm （1,680 bp）。 phaC1Pm 和phaC2Pm 推导的氨基酸序列与其他假单胞菌的相应PHA合酶具有最高的相似性。这两个PHA合酶基因使PHA阴性突变体恶臭假单胞菌GPp104和富营养小球藻PHB-4合成PHA。 4。在恶臭假单胞菌GPp104中，由phaC1Pm 和phaC2Pm 介导的中等链长羟基链烷酸酯（C6-C12）的PHA合成，这在其他假单胞菌中经常报道。相比之下，在富营养R. PHB–4中，任一PHA合酶基因也导致3-羟基丁酸酯（3HB）掺入PHA。带有门氏假单胞菌phaC基因的富营养R. eutropha PHB–4重组菌株在葡萄糖酸盐培养过程中甚至积累了3HB的均聚酯，如果phaC2为p3，则poly（3HB）占细胞干物质的80％以上。表达。有趣的是，当用脂肪酸作为唯一碳源培养时，携带phaC1合酶基因的重组细胞积累较高的PHA，而当在任一宿主中将葡萄糖酸盐用作碳源时，携带PhaC2合酶的重组细胞积累较高的PHA。此外，P.mendocina的等基因phaC1和phaC2敲除突变体提供了证据，证明PhaC1是P.mendocina中PHA合成的主要酶，而PhaC2仅在很小的程度上促进了PHA在该细菌中的积累。当在葡萄糖酸盐上培养时。

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  • 来源
    《Applied Microbiology and Biotechnology》 |2002年第2期|229-236|共8页
  • 作者

    S. Hein; J. Paletta; A. Steinbüchel;

  • 作者单位

    Institut für Mikrobiologie der Westfälischen Wilhelms-Universität Münster Corrensstrasse 3 48149 Münster Germany;

    Institut für Mikrobiologie der Westfälischen Wilhelms-Universität Münster Corrensstrasse 3 48149 Münster Germany;

    Institut für Mikrobiologie der Westfälischen Wilhelms-Universität Münster Corrensstrasse 3 48149 Münster Germany;

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