马克斯克鲁维酵母NBRC1777来源的β-葡萄糖苷酶的纯化、结晶及X射线初步分析
Purification, crystallization and preliminary X-ray analysis of beta-glucosidase from Kluyveromyces marxianus NBRC1777.
作者信息
Yoshida Erina, Hidaka Masafumi, Fushinobu Shinya, Koyanagi Takashi, Minami Hiromichi, Tamaki Hisanori, Kitaoka Motomitsu, Katayama Takane, Kumagai Hidehiko
机构信息
Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi, Ishikawa, Japan.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Nov 1;65(Pt 11):1190-2. doi: 10.1107/S1744309109042948. Epub 2009 Oct 30.
Abstract
The intracellular beta-glucosidase from Kluyveromyces marxianus NBRC1777 (KmBglI) belongs to glycoside hydrolase family 3 and has a unique domain architecture. Selenomethionine-labelled KmBglI was purified and crystallized by the hanging-drop vapour-diffusion method using the purified enzyme at 30 mg ml(-1), 0.04 M potassium dihydrogen phosphate pH 5.1, 16%(w/v) PEG 8000 and 20%(v/v) glycerol. The crystal belonged to space group C2, with unitcell parameters a = 245.8, b = 148.7, c = 119.9 angstrom, beta = 112.9 degrees. Multiple-wavelength anomalous dispersion data were collected at 2.4 and 2.5 angstrom resolution. A tetramer was assumed to be present in the asymmetric unit, which gave a Matthews coefficient of 2.6 angstrom(3) Da(-1).
摘要
来自马克斯克鲁维酵母NBRC1777（KmBglI）的细胞内β-葡萄糖苷酶属于糖苷水解酶家族3，具有独特的结构域结构。使用浓度为30 mg/ml的纯化酶、0.04 M磷酸二氢钾（pH 5.1）、16%（w/v）聚乙二醇8000和20%（v/v）甘油，通过悬滴气相扩散法对硒代甲硫氨酸标记的KmBglI进行纯化和结晶。晶体属于空间群C2，晶胞参数a = 245.8、b = 148.7、c = 119.9埃，β = 112.9°。在2.4和2.5埃分辨率下收集了多波长反常色散数据。假定不对称单元中存在一个四聚体，其马修斯系数为2.6埃³ Da⁻¹。
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