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来自大麻的聚酮合酶-1(PKS-1)的结晶及初步X射线衍射研究。

Crystallization and preliminary X-ray diffraction studies of polyketide synthase-1 (PKS-1) from Cannabis sativa.

作者信息

Taguchi Chiho, Taura Futoshi, Tamada Taro, Shoyama Yoshinari, Shoyama Yukihiro, Tanaka Hiroyuki, Kuroki Ryota, Morimoto Satoshi

机构信息

Faculty of Pharmaceutical Sciences, Kyushu University, Japan.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Mar 1;64(Pt 3):217-20. doi: 10.1107/S1744309108003795. Epub 2008 Feb 29.

Abstract

Polyketide synthase-1 (PKS-1) is a novel type III polyketide synthase that catalyzes the biosynthesis of hexanoyl triacetic acid lactone in Cannabis sativa (Mexican strain). PKS-1 was overproduced in Escherichia coli, purified and finally crystallized in two different space groups. The crystal obtained in 0.1 M HEPES buffer pH 7.5 containing 0.2 M calcium acetate and 20%(w/v) polyethylene glycol 3350 diffracted to 1.65 A resolution and belonged to space group P1, with unit-cell parameters a = 54.3, b = 59.3, c = 62.6 A, alpha = 69, beta = 81, gamma = 80 degrees. Another crystal obtained in 0.1 M HEPES buffer pH 7.5 containing 0.2 M sodium chloride and 20%(w/v) polyethylene glycol 3350 diffracted to 1.55 A resolution and belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 54.3, b = 110, c = 130 A. These data will enable us to determine the crystal structure of PKS-1.

摘要

聚酮合酶-1(PKS-1)是一种新型的III型聚酮合酶,可催化大麻(墨西哥品种)中己酰三乙酸内酯的生物合成。PKS-1在大肠杆菌中过量表达,经过纯化,最终在两种不同的空间群中结晶。在含有0.2 M乙酸钙和20%(w/v)聚乙二醇3350的pH 7.5的0.1 M HEPES缓冲液中获得的晶体,其衍射分辨率为1.65 Å,属于空间群P1,晶胞参数为a = 54.3、b = 59.3、c = 62.6 Å,α = 69°、β = 81°、γ = 80°。在含有0.2 M氯化钠和20%(w/v)聚乙二醇3350的pH 7.5的0.1 M HEPES缓冲液中获得的另一种晶体,其衍射分辨率为1.55 Å,属于空间群P2(1)2(1)2(1),晶胞参数为a = 54.3、b = 110、c = 130 Å。这些数据将使我们能够确定PKS-1的晶体结构。

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Crystallization of Delta1-tetrahydrocannabinolic acid (THCA) synthase from Cannabis sativa.来自大麻的Δ1-四氢大麻酚酸(THCA)合酶的结晶。
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