铜绿假单胞菌的FabG是一种依赖NADPH的3-酮酰基还原酶，可在大肠杆菌中为中链长度聚3-羟基链烷酸酯的生物合成提供前体。
FabG, an NADPH-dependent 3-ketoacyl reductase of Pseudomonas aeruginosa, provides precursors for medium-chain-length poly-3-hydroxyalkanoate biosynthesis in Escherichia coli.
作者信息
Ren Q, Sierro N, Witholt B, Kessler B
机构信息
Institute of Biotechnology, ETH Hönggerberg, CH-8093 Zürich, Switzerland.
出版信息
J Bacteriol. 2000 May;182(10):2978-81. doi: 10.1128/JB.182.10.2978-2981.2000.
Abstract
Escherichia coli hosts expressing fabG of Pseudomonas aeruginosa showed 3-ketoacyl coenzyme A (CoA) reductase activity toward R-3-hydroxyoctanoyl-CoA. Furthermore, E. coli recombinants carrying the poly-3-hydroxyalkanoate (PHA) polymerase-encoding gene phaC in addition to fabG accumulated medium-chain-length PHAs (mcl-PHAs) from alkanoates. When E. coli fadB or fadA mutants, which are deficient in steps downstream or upstream of the 3-ketoacyl-CoA formation step during beta-oxidation, respectively, were transformed with fabG, higher levels of PHA were synthesized in E. coli fadA, whereas similar levels of PHA were found in E. coli fadB, compared with those of the corresponding mutants carrying phaC alone. These results strongly suggest that FabG of P. aeruginosa is able to reduce mcl-3-ketoacyl-CoAs generated by the beta-oxidation to 3-hydroxyacyl-CoAs to provide precursors for the PHA polymerase.
摘要
表达铜绿假单胞菌fabG的大肠杆菌宿主对R-3-羟基辛酰辅酶A显示出3-酮酰基辅酶A（CoA）还原酶活性。此外，除fabG外还携带聚3-羟基链烷酸酯（PHA）聚合酶编码基因phaC的大肠杆菌重组体从链烷酸积累中链长度的PHA（mcl-PHA）。当分别在β-氧化过程中3-酮酰基辅酶A形成步骤的下游或上游步骤存在缺陷的大肠杆菌fadB或fadA突变体用fabG转化时，与仅携带phaC的相应突变体相比，大肠杆菌fadA中合成的PHA水平更高，而在大肠杆菌fadB中发现的PHA水平相似。这些结果有力地表明，铜绿假单胞菌的FabG能够将β-氧化产生的中链3-酮酰基辅酶A还原为3-羟基酰基辅酶A，为PHA聚合酶提供前体。
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