6533b822fe1ef96bd127d98f
RESEARCH PRODUCT
Expression, purification, crystallization and preliminary X-ray analysis of strictosidine glucosidase, an enzyme initiating biosynthetic pathways to a unique diversity of indole alkaloid skeletons
Guohong PengJoachim StöckigtJuergen KoepkeHartmut MichelLeif BarlebenXueyan Masubject
Ammonium sulfateCatharanthusStereochemistryBiophysicsCrystallography X-Raymedicine.disease_causeBiochemistryIndole AlkaloidsAnalytical Chemistrychemistry.chemical_compoundRauvolfia serpentinaPEG ratioEscherichia colimedicineCloning MolecularMolecular BiologyEscherichia colichemistry.chemical_classificationbiologyIndole alkaloidbiology.organism_classificationEnzymeBiochemistrychemistryStrictosidineCrystallizationSodium acetateGlucosidasesdescription
Abstract Strictosidine β- 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 P 42 1 2 with unit cell dimensions of a =157.63, c =103.59 A and diffract X-rays to 2.48-A resolution.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2005-02-01 | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics |