Chimeric Enzyme Composed of Polyhydroxyalkanoate (PHA) Synthases from Ralstonia eutropha and Aeromonas caviae Enhances Production of PHAs in Recombinant Escherichia coli
Chimeric Enzyme Composed of Polyhydroxyalkanoate (PHA) Synthases from Ralstonia eutropha and Aeromonas caviae Enhances Production of PHAs in Recombinant Escherichia coli
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DOI：
10.1021/bm801386j
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发表时间：
2009-04-01
影响因子：
6.2
通讯作者：
Taguchi, Seiichi
中科院分区：
文献类型：
--
作者：
Matsumoto, Ken'ichiro;Takase, Kazuma;Taguchi, Seiichi
构建了由真核梭菌(Cupriavidus Eiator)聚羟基烷酸(PHA)合成酶(PHAc(Re))和豚鼠气单胞菌(Aeromonas caviae，PHAc(Ac))聚羟基烷酸(PHA)合成酶组成的嵌合酶。PHAC(Re)是已知的具有特征的PHA合成酶中具有很强的酶活性的酶。PHAC(Ac)具有广泛的底物专一性，可合成短链(SCL)/中链(MCL)PHA。我们试图创造继承这两种有利性质的嵌合酶。在8个嵌合体中，Acre12(PhaCA的N端占26%，Phac(Re)的C端占74%)在大肠杆菌JM109中表现出与亲本酶相似的P(3-羟基丁酸)积累。因此，将AcRe12应用于以E.coliLS5218为宿主的SCL/MCL PHA生产中。AcRe12的PHA积累量(50wt%)高于亲本酶。此外，PHA由2摩尔%3-羟基己酸酯和3-羟基丁酸酯组成。因此，嵌合的PHA合成酶Are12继承了这两种亲本酶的特性，从而表现出更好的酶性质。
Chimeric enzymes composed of polyhydroxyalkanoate (PHA) synthases from Ralstonia eutropha (Cupriavidus necator) (PhaC(Re)) and Aeromonas caviae (PhaC(Ac)) were constructed. PhaC(Re) is known for its potent enzymatic activity among the characterized PHA synthases. PhaC(Ac) has broad substrate specificity and synthesizes short-chain-length (SCL)/medium-chain-length (MCL) PHA. We attempted to create chimeric enzymes inheriting both of the advantageous properties. Among eight chimeras, AcRe 12, with 26% of the N-terminal of PhaCA, and 74% of the C-terminal of PhaC(Re), exhibited comparable P(3-hydroxybutyrate) accumulation as parental enzymes in Escherichia coli JM109. Thus, AcRe12 was applied to SCL/MCL PHA production using E. coli LS5218 as the host. AcRe12 accumulated higher amount of PHA (50 wt %) than the parental enzymes. Furthermore, the PHA consisted of 2 mol % 3-hydroxyhexanoate as well as 3-hydroxybutyrate. Therefore, the chimeric PHA synthase, AcRe 12, inherited the character of both of the parental enzymes and thus exhibits improved enzymatic properties.