Crystallization and preliminary X-ray analysis of (R)-specific enoyl-CoA hydratase from Aeromonas caviae involved in polyhydroxyalkanoate biosynthesis.
Crystallization and preliminary X-ray analysis of (R)-specific enoyl-CoA hydratase from Aeromonas caviae involved in polyhydroxyalkanoate biosynthesis.
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来自豚鼠气单胞菌参与聚羟基链烷酸酯生物合成的 (R) 特异性烯酰辅酶 A 水合酶的结晶和初步 X 射线分析。
DOI：
10.1107/s0907444900014062
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Y. Doi
中科院分区：
文献类型：
--
作者：
T. Hisano;T. Fukui;T. Iwata;Y. Doi
来自豚鼠气单胞菌的二聚体 (R) 特异性烯酰辅酶 A (CoA) 水合酶可催化碳长度为 4-6 的反式-2-烯酰辅酶 A 水合，产生相应的 (R)-3-羟基酰基-CoA，对于聚羟基脂肪酸酯 (PHA) 生物合成至关重要。该酶在 298 K 下通过蒸气扩散在含有 20% 聚乙二醇 4000、5% 2-丙醇和 20 mM HEPES pH 7.0 的储存溶液中结晶。晶体属于单斜空间群 C2，晶胞参数 a = 111.54 (3)、b = 59.29 (1)、c = 47.27 (4) A、beta = 113.04(2)度且在不对称单元中含有二聚体分子。晶体在 100 K 下的闪蒸冷却将其晶胞参数更改为 a = 109.82 (7)、b = 57.98 (6)、c = 46.84 (2) A、beta = 112.71 (3) 度。使用同步加速器辐射在低温 (100 K) 条件下收集的原始数据分辨率为 1.7 A，完整性为 94.5%，R（合并）为 4.0%。
Dimeric (R)-specific enoyl-coenzyme A (CoA) hydratase from Aeromonas caviae catalyzes the hydration of trans-2-enoyl-CoAs with carbon lengths of 4-6 to yield their corresponding (R)-3-hydroxyacyl-CoAs and is essential for polyhydroxyalkanoate (PHA) biosynthesis. The enzyme has been crystallized by vapour diffusion against a reservoir solution containing 20% polyethylene glycol 4000, 5% 2-propanol and 20 mM HEPES pH 7.0 at 298 K. Crystals belong to the monoclinic space group C2, with unit-cell parameters a = 111.54 (3), b = 59.29 (1), c = 47.27 (4) A, beta = 113.04 (2) degrees and contain a dimeric molecule in the asymmetric unit. Flash-cooling of a crystal at 100 K alters its unit-cell parameters to a = 109.82 (7), b = 57.98 (6), c = 46.84 (2) A, beta = 112.71 (3) degrees. Native data to a resolution of 1.7 A have been collected with 94.5% completeness and an R(merge) of 4.0% under cryogenic (100 K) conditions using synchrotron radiation.