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Report No.
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Crystallization and preliminary X-ray diffraction studies of polyketide synthase-1 (PKS-1) from ${it Cannabis sativa}$

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

Polyketide synthase-1 (PKS-1) is a novel type III polyketide synthase that catalyzes biosynthesis of the hexanoyl triacetic acid lactone in ${it Cannabis sativa}$ (Mexican strain). PKS-1 was overproduced in ${it Escherichia coli}$, 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 ${AA}$ resolution and belonged to space group ${it P}$1 with unit-cell parameters of a = 54.3 ${AA}$, b = 59.3 ${AA}$, c = 62.6 ${AA}$, ${it $alpha$}$ = 69$$^{circ}$$, ${it $beta$}$ = 81$$^{circ}$$, ${it $gamma$}$ = 80$$^{circ}$$. 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 ${AA}$ resolution and belonged to the space group ${it P}$2$$_{1}$$2$$_{1}$$2$$_{1}$$ with unit-cell parameters of a = 54.3 ${AA}$, b = 110 ${AA}$, c = 130 ${AA}$. These data will enable us to determine the crystal structure of PKS-1.

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Category:Biochemical Research Methods

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