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首页> 外文期刊>Applied Microbiology >Wide Distribution among Halophilic Archaea of a Novel Polyhydroxyalkanoate Synthase Subtype with Homology to Bacterial Type III Synthases
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Wide Distribution among Halophilic Archaea of a Novel Polyhydroxyalkanoate Synthase Subtype with Homology to Bacterial Type III Synthases

机译：嗜盐古细菌之间的广泛分布的一种新的多羟基链烷酸合酶亚型与细菌III型合酶的同源性。

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Polyhydroxyalkanoates (PHAs) are accumulated as intracellular carbon and energy storage polymers by various bacteria and a few haloarchaea. In this study, 28 strains belonging to 15 genera in the family Halobacteriaceae were investigated with respect to their ability to synthesize PHAs and the types of their PHA synthases. Fermentation results showed that 18 strains from 12 genera could synthesize polyhydroxybutyrate (PHB) or poly(3-hydroxybutyrate- co -3-hydroxyvalerate) (PHBV). For most of these haloarchaea, selected regions of the phaE and phaC genes encoding PHA synthases (type III) were cloned via PCR with consensus-degenerate hybrid oligonucleotide primers (CODEHOPs) and were sequenced. The PHA synthases were also examined by Western blotting using haloarchaeal Haloarcula marismortui PhaC (PhaC_(Hm)) antisera. Phylogenetic analysis showed that the type III PHA synthases from species of the Halobacteriaceae and the Bacteria domain clustered separately. Comparison of their amino acid sequences revealed that haloarchaeal PHA synthases differed greatly in both molecular weight and certain conserved motifs. The longer C terminus of haloarchaeal PhaC was found to be indispensable for its enzymatic activity, and two additional amino acid residues (C143 and C190) of PhaC_(Hm) were proved to be important for its in vivo function. Thus, we conclude that a novel subtype (IIIA) of type III PHA synthase with unique features that distinguish it from the bacterial subtype (IIIB) is widely distributed in haloarchaea and appears to be involved in PHA biosynthesis.
机译：聚羟基链烷酸酯（PHA）被各种细菌和一些盐生古细菌作为细胞内碳和能量存储聚合物积累。在这项研究中，研究了盐杆菌科15个属的28个菌株的合成PHAs的能力及其PHA合成酶的类型。发酵结果表明，来自12个属的18个菌株可以合成聚羟基丁酸酯（PHB）或聚（3-羟基丁酸酯-co-3-羟基戊酸酯）（PHBV）。对于大多数这些古细菌，编码的PHA合成酶（III型）的phaE和phaC基因的选定区域通过使用共有简并杂交杂合寡核苷酸引物（CODEHOP）进行PCR克隆并进行了测序。 PHA合成酶也用西方古细菌Halocrcula marismortui PhaC（PhaC_（Hm））抗血清通过蛋白质印迹法检查。系统发育分析表明，来自嗜盐菌科和细菌域的III型PHA合成酶分别聚集。比较它们的氨基酸序列发现，卤代古菌PHA合成酶在分子量和某些保守基序上都有很大差异。发现卤代古细菌PhaC的更长的C末端对于其酶活性是必不可少的，并且已证明PhaC_（Hm）的另外两个氨基酸残基（C143和C190）对其体内功能很重要。因此，我们得出结论，III型PHA合酶的新亚型（IIIA）具有独特的特征，可与细菌亚型（IIIB）区别开来，广泛分布于盐生古细菌中，似乎参与了PHA生物合成。

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