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Expression, purification, crystallization and preliminary X-ray analysis of strictosidine glucosidase, an enzyme initiating biosynthetic pathways to a unique diversity of indole alkaloid skeletons

被引：32
作者：
Barleben, L
Ma, XY
Koepke, J
Peng, GH
Michel, H
Stöckigt, J
机构：
[1] Johannes Gutenberg Univ Mainz, Inst Pharm, Dept Pharmaceut Biol, D-55099 Mainz, Germany
[2] Max Planck Inst Biophys, Dept Mol Membrane Biol, D-60439 Frankfurt, Germany
来源：
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS | 2005年 / 1747卷 / 01期
关键词：
strictosidine beta-D-glucosidase; cloning and purification; crystallization; X-ray analysis; Rauvoltia serpentina;
D O I：
10.1016/j.bbapap.2004.09.026
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Strictosidine beta-D-glucosidase, a plant enzyme initiating biosynthetic pathways to about 2000 monoterpenoid indole alkaloids with an extremely large number of various carbon skeletons, has been functionally expressed in Escherichia coli and purified to homogeneity in mg scale. Crystals suitable for X-ray analysis were found by robot-mediated screening. Using the hanging-drop technique, optimum conditions were 0.3 M ammonium sulfate, 0.1 M sodium acetate, pH 4.6 and PEG 4000 (10%) as precipitant buffer. The crystals of strictosidine glucosidase belong to the space group P42(1)2 with unit cell dimensions of a=157.63, c=103.59 Angstrom and diffract X-rays to 2.48-Angstrom resolution. (C) 2004 Elsevier B.V All rights reserved.
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页码：89 / 92
页数：4
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