title: Expression, purification, crystallization and preliminary X-ray analysis of strictosidine glucosidase, an enzyme initiating biosynthetic pathways to a unique diversity of indole alkaloid skeletons.
abstract: 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 A and diffract X-rays to 2.48-A resolution.
authors.affiliation: Department of Pharmaceutical Biology, Institute of Pharmacy, Johannes Gutenberg-University Mainz, Staudinger Weg 5, D-55099 Mainz, Germany.
authors.forename: Leif
authors.initials: L
authors.lastname: Barleben
authors.forename: Xueyan
authors.initials: X
authors.lastname: Ma
authors.forename: Juergen
authors.initials: J
authors.lastname: Koepke
authors.forename: Guohong
authors.initials: G
authors.lastname: Peng
authors.forename: Hartmut
authors.initials: H
authors.lastname: Michel
authors.forename: Joachim
authors.initials: J
authors.lastname: Stöckigt
mesh_terms: D030008:Catharanthus / Q000201:enzymology*; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005959:Glucosidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D026121:Indole Alkaloids / Q000378:metabolism*
chemical_list: D026121:Indole Alkaloids; D005959:Glucosidases; C456847:strictosidine glucosidase